07-27-01 05:48
No 199197
This is a work in progress. It’s by Mobius Bandsaw
At present I have a fairly good grasp of the means and the end.
You are invited and encouraged to contribute information you may have relating to this subject, we can work together to advance the cause of knowledge.
Shall we begin?
Forward:
When I was given this forum I was also given something of a research assignment.
It seems there is a great deal of interest in the production of ephedrine and pseudoephedrine. (The forum was called “Homebrewing”)
Both of these products may be obtained in quantity using methods which are in normal practice in industry.
Industrial production of L-PACs uses methodologies which are not altogether removed from the art of distillers of spirits.
While still speaking about theoretical methods of production of alcohols, and products of alcohols, it is hoped that a practical method of production of ephedrine and pseudoephedrine will be achieved which could be employed by the individual
should circumstance find him in need of such capability.
It should be noted from the outset that these methods, recipies, informations, and observations, are not intended to be used by any person.
Further, all persons are hereby advised that use of this information to produce any substance for any illegal purpose is forbidden, and that the author expressly withdraws any permission to read, store, record, or in any way archive this work,
from any person, persons, entity, or organisation, intending to use this information for any illegal purpose.
Any methods discussed here are purely speculative, and for informative and entertainment purposes only, and have not been attempted.
Untrained persons working with devices and substances mentioned here risk serious injury or death.
Production by unlicensed individuals of any substance mentioned or described here could lead to criminal prosecution.
~~~~~~~~~~~~~~~~~~~ 1.~~~~~~~~~~~~~~~~~~~~~
FUN WITH L-PAC production!
Biotransformation processes involving both yeast (Candida utilis) and pyruvate decarboxylase (PDC) for the production of L-phenylacetylcarbinol (L-PAC) from substrates benzaldehyde and pyruvate are an interesting field of study!
L-PAC is an intermediate in the production of ephedrine and pseudoephedrine.
Models for the process with a substrate feeding profile for benzaldehyde will be proposed.
Chiral vicinal aminoalcohols are key building blocks for the production of a number of pharmaceutical products.
Important compounds which may be produced from chiral vicinal aminoalcohol precursors span a range of therapeutic categories and include pseudoephedrine.
Patent US5834261 describes a process that may be employed to produce a broad range of chiral vicinal aminoalcohols, both cyclic and acyclic.
A particularly notable feature of this method is the ability to control the absolute configuration at chiral centers bearing both the amino and alcohol functionality to produce any of the four possible stereoisomers in high stereochemical purity.
Patent US5942644 (issued in August 1999), is related to the process patent described above.
This second patent covers the key intermediates that are involved in the production of chiral vicinal aminoalcohols. These compounds include the hydrazides and hydroxamic acids that are the immediate precursors of chiral vicinal aminoalcohols.
The process for producing chiral vicinal aminoalcohols relies on readily available þ-ketoesters as starting materials.
Examples of þ-ketoesters useful for the production of chiral vicinal aminoalcohols include the inexpensive compounds acetoacetic ester and þ-keto-phenylpropionic acid esters.
Central to this method for the production of these chiral vicinal aminoalcohols is the combination of two key steps, each of which proceeds with a well-defined and controllable stereochemical outcome.
The first step is the stereoselective reduction of the keto group of a þ-ketoester to produce the corresponding þ-hydroxyester.
This reaction is catalyzed by an alcohol dehydrogenase in the presence of a nicotinamide cofactor.
Because of the facile equilibrium between the two enantiomers of a 2-substituted-þ-ketoester in aqueous solution, the interconversion of these two stereoisomers occurs rapidly.
The reduction of the ketone by an alcohol dehydrogenase occurs with a high degree of stereoselectivity, reducing only one of the two ketone enantiomers.
The reduction of the ketone is highly stereoselective for the production of a single alcohol stereoisomer.
so, two chiral centers are generated simultaneously by this enzymatic reaction, and this reaction provides for control of stereochemistry at both the C-2 and C-3 positions of the 2-substituted-þ-ketoester.
NEXT: How earth people do it.
~~~~~~~~~~HOW EARTH PEOPLE DO IT~~~~~~~~~~~~
If your nitemares never include metalic voices coming over loudspeakers
demanding that you throw out your five gallon plastic buckets and come out with
your hands up, congratulations...you may be on Earth.
Earth people find many wonderful uses for the humble five gallon plastic bucket, as we shall soon see.
Any earth native who wishes to produce L-PAC at home may discover these utilitarian devices to be worth well more than their wieght in gold.
To produce L-PAC using biosynth methods, one must first decide what will be fermented in the buckets.
In the course of my research I have discovered that the liquid obtained by the crushing, grinding, pressing, and filtering of the common sugar beet is in fact the best possible substance to use for this method due to natural enzyme feeding, but it
is by no means the only one, according to everyones favorite Uncle, even water may be used as a starting point!
After obtaining the liquid to be used, one may wish to fortify it with a bit of brewers sugar.
Into the 4.5 gallons of liquified beet extract one may add about 5 lbs of brewers sugar until the sugar no longer wishes to disolve, the reason for this being the already high sugar content of the extract.
The next requirement is to add yeast. Not just any yeast will do.
For our purposes the strain named Candida Utilis is understood to be best, however, experimentation is encouraged.
Candida Utilis enjoys wide popularity, and may easily be obtained.
The proper addition of yeast is a point of contention.
Many "experts" say that one adds only a small amount, such as say a couple of packets of brewers yeast.
Others state that it is necessary to put in an amount of wieght equal to the added sugar. It would seem to me that a benefit of doing this would be that the process will progress much quicker (three or four days, as opposed to three or four weeks) if
one were to add the larger amount of yeast.
Yeast is expensive if you buy it one package at a time.
Most brewers know how to increase yeast.
Those who do not are again encouraged to purchase a brewers manual detailing the method.
Another required substance is Benzaldehyde.
The production of L-PAC will be directly proportional to the amount of Benzaldehyde used. Returns are expected to be in the range of 60 to 80% the wieght of the added Benzaldehyde.
Once the needed items have been procured one may begin the progress of biosynthesis.
Step one involves mixing the nutrient, which is the liquid, and sugar. Next one adds the yeast to the mixture.
Be careful to avoid contamination of your mixture!
After combining the ingredients, place the top loosely on the five gallon bucket. With the passage of an hour or so one may begin to see the process of fermentation begining, it is felt that the passage of at least 10 hours is needed to allow the fermentation process to fully become active.
After ten hours one must add the benzaldehyde. For this feeding profile the proper amount is calculated to be 60ML.
After adding the Benzaldehyde the fermentation should be left to progress in a cool place away from sunlight. While underway this process also needs to be free of physical shock, such as thumps, and quaking, since this retards the process. If you have a Rock and Roll band and practice in the garage, don't do your biosynth there.
NEXT: IT WORKED! NOW WHAT?
~~~~~~~~~IT WORKED! NOW WHAT?~~~~~~~~~~~~~
Now that your fermentation has completed you have your precurson to ephedrine just floating around in your five gallon bucket.
It's not doing you much good in there is it?
The next thing you want to do is to recover your phenylpropanol which is what the yeast turned the sugars into with the help of the benzaldehyde.
The most direct method of recovery is to use a centrifuge, but few of us have one on hand. A more practical method for the small scale producer is filtration.
Now that you have your filtered liquid it is time to begin extracting the phenylpropanol.
To extract the phenylpropanol, you will need to use a non polar solvent, here you may choose between Acetone, Tolulene, or Xylene (personal preference here is toward acetone).
The about 200ml of NP should be added to the mixture, swirled, then allowed to seperate, then decanted, save it, your goodies are there. Repeat this proceedure three times.
At the end of the third wash you should have your phenylpropanol in solvent.
Next you will need to distill the solvent/phenylpropanol in order to remove the solvent.
Once your solvent is largely gone you will again need to distill, but this time you will need to use vacuum in your distillation process. You will need to pull between 14 and 18 torr and your product will come over as phenylacetylcarbinol at between 105c and 155c.
All that remains to be done is a fairly standard reductive amination, whereupon you will find yourself in possession of pure gak free ephedrine.
~~~~~~~~~~~~~~~~~~~NOTE~~~~~~~~~~~~~~~~~
Comments, suggestions, and refinements are all solicited.
~~~~~~~~~~~~~~~REFERENCES~~~~~~~~~~~~~~~
Uncle Fester, SOMM
Shin, H.S. and Rogers, Production of L-PAC from benzaldehyde using partially purified pyruvate decarboxylase.
Rogers, P.L., Shin, H.S. and Wang, Biotransformation for L-ephredrine production.
Benzaldehyde: Artificial essential oil of almond; artificial almond flavor is water,
benzaldehyde and alcohol.
Occurs in kernels of bitter almonds; made synthetically from benzal chloride (C6H5CHCl2)
and lime, or by oxidation of toluene.
Hydrolysing benzal chloride is no challenge, boiling in alkaline water. It's making it, by
chlorinating toluene, and keeping it separated from benzyl chloride, the monochloro
derivative, that's the challenge.
In toluene oxidation, the challenge is to prevent overoxidation to benzoic acid. My ballot
is marked for acidic manganese dioxide, but I have seen chromyl chloride mentioned.
A convenient natural source of benzaldehyde is by hydrolysis of amygdalin. That spelling
challenge is a double glucoside of mandelonitrile, benzaldehyde's cyanide derivative.
Mostly in apricot pits and bitter almonds, also the seeds of peaches and cherries.
In these natural sources is some free benzaldehyde, but most of it is locked in the
amygdalin. But there is an enzyme in the natural seeds, which will hydrolyse the
amygdalin for you, when you let the ground seed material stand in water; glucose,
benzaldehyde, and hydrogen cyanide result.
Posted by--anomolon
This is a link you might want to look at: http://www.geocities.com/dritte123/ephye
Be sure to look up these two patents, Patent US1956950 & Patent US1962476.
My intent on posting this information is for others to join in on the positive and see if we as a whole could bring this to a standard in our quest to rid our self’s of the dependance of a pill to produce a product. It should be looked at and responded to in that light... one bee of the Hive, Jacked
(Ancient Alchemist Delux)
07-27-01 16:38
No 199320
Could this thread be moved to the serious chemistry forum. It was a mistake placing it
here I believe.
Paid in Full
(Stranger)
07-29-01 04:53
No 199661
Swish is very happy that someone was able to rescue this post from the Zonez. Swish has full intention of giving this some serious experimentation when everything is gathered.
(Newbee)
08-20-01 23:22
No 206068
This is veerrrrrrry interesting!
I have just a couple of Qs...1) You suggest using acetone as the non-polar solvent... wouldnt acetone bee miscible with the sugar beet juice?
2) You use 200mL solvent for each extraction - isnt this far too little for 4.5gallons of solution?
3) Would this work for substituted benzaldehydes?
4) Is it really necessary to vac-distil the L-PAC before doing a reductive amination?
Many thanks,
Ololiuqui
(Chief Bee)
08-21-01 00:59
No 206116
I have heard that the yields are very low with substituted benzaldehydes.
(Hive Bee)
08-22-01 02:36
No 206424
I refer you to this tid bit of info.
__________________________________
!!!! I FORGOT TO QUOTE THAT THIS REACTION IS STRONGLY pH DEPENDENT AND
BEST PROCEEDS AT A pH OF 8-9. !!!!!
From Agric. Biol. Chem., Volume 50, yr 1986, page 1261
"Reductive C3-Homologation of Substituted Benzaldehydes by
Fermenting Bakers' Yeast
...
... A unique reaction catalyzed by Bakers' Yeast is the reductive C-C bond formation in
benzaldehyde that results in the formation of 1-phenylpropane-1,2-diol [ref 3]. This reaction is
supposed to involve the attack of pyruvic acid, which is formed from sugars through a well known
pathway, by benzaldhyde to give phenyl-1-hydroxy-2-propanone [ref 4]. This ketol is considered to
be reduced by alcohol dehydrogenase of the yeast to give the
diol. ...
...
Reduction of aldehydes by fermenting yeast. General procedure. To 50 mL of tap water, 12.5 g
of dry yeast and 10 g glucose were added, followed by stirring
at room temperature for 10 minutes. Then a solution of aldehyde 1 (about 600 mg) in 1 mL of
ethanol was added to the suspension of fermenting yeast with stirring. After 1 hour yeast (6.2 g)
and glucose (5 g) were added to the reaction mixture, followed by stirring for an additional 3.5
hours. The reaction mixture was then poured into a beaker containing 35 grams of celite and
extracted with 100 mL of ethyl acetate three times. The combined organic layer was dried over
anhydrous Na2SO4. The solvent removed under reduced pressure to give an oil consisting of
benzylic alcohol 3, 1-arylpropane-1,2 diol 3, and in
some cases, 1-aryl-1-hydroxy-2-propanone 4. ...
Table II. Yields and Steroselectivities of propanediols (a)
________________________________________
Substrate______Yield %____...
________________________________________
H______________30
p-OMe__________22
p-Me____________28
p-Cl____________27
p-F_____________26
p-NO2__________14
o-Me____________7
o-Cl_____________32
o-F_____________30
m-F____________31
CF3(e)__________0
_________________________
(a) Reactions were carried out in tap water at room
temperature ... (e) All of the o-, m-
p-substituted isomers gave no diols."
_________________
If i remember correctly, the diol can be rearranged to form P2P with H2SO4
(Ancient Alchemist Delux)
08-22-01 11:45
No 206519
You suggest using acetone as the non-polar solvent... wouldnt acetone bee
miscible with the sugar beet juice?
At the point in the procedure that the acetone is used, the sugar beet
juice has been fermented.
The solvent will collect what it is after, and seperate (layer).
You use 200mL solvent for each extraction - isnt this far too little for 4.5gallons of solution?
Most of the 4.5 gallons is junk that you are not interested in. The 200Ml
per extraction is conservative, you may use more if you wish, the only
complication is the added seperation/evaporation time.
Would this work for substituted benzaldehydes?
It would depend on the substitution
Is it really necessary to vac-distil the L-PAC before doing a reductive
amination?
Yes. Without the vacuum distillation you will not be aminating only the
desired substance. I am unsure what contaminates may reside in the
undistilled form. For safety's sake, I would not skip this step.
Answers were given to these questions by the writer of the original post.
http://crystalninjas.net/jacked/disclaim
(Stranger)
08-22-01 19:07
No 206566
In your quotation from the journal you say:
The solvent removed under reduced pressure to give an oil consisting of
benzylic alcohol 3, 1-arylpropane-1,2 diol 3, and in
some cases, 1-aryl-1-hydroxy-2-propanone 4. ...
What is the 4. part of the last word? I do not want to assume anything!!!
It also appears that the large excess of yeast is used to allow immediate gratification to the above mentioned experimentalists!
I dunno, but I been told ... You never slow down, you never grow old!
(Hive Bee)
08-23-01 04:03
No 206663
To clarify:
"... benzylic alcohol 3, 1-arylpropane-1,2 diol 3, and in
some cases, 1-aryl-1-hydroxy-2-propanone 4. ..."
the 3, and 4 refer drawings of the compounds on diagram not included.
(Official Hive Translator)
09-19-01 19:14
No 215161
....having read the proc. about getting -propanediol from benzaldehyde, i couldn't help wondering about 2 things:
1) The procedure doesn't mention any sort of pH adjustment - how is pH supposed to get to 9? It also just looks strange - what is the origin of that 1st phrase, the one in CAPS? Jim, you still around? Please, explain!
2) Apart from the pH thing, the whole procedure seems completely identical to l-PAC production!! What is you think that makes the difference?
Antoncho will bee VERY appreciative of any help on this issue, cause SW He Doesn't Even Know just happens to have a kilo of benzaldehyde lying around with no use for it :):):)
Thank you all in advance,
impatiently awaiting
Antoncho
P.S. Dear Moderators, do you really think this thread belongs here? - if mr. Jacked doesn't mind...
(Distinctive Doe)
09-20-01 05:05
No 215347
*****ADVANCES IN BIOCHEMICAL ENGINEERING/BIOTECHNOLOGY*****
Rogers PL Shin HS Wang B
Biotransformation for L-ephedrine production.
In: Adv Biochem Eng Biotechnol (1997) 56:33-59
ISSN: 0724-6145
L-ephedrine is widely used in pharmaceutical preparations as a decongestant and
anti-asthmatic compound. One of the key intermediates in its production is
L-phenylacetylcarbinol (L-PAC) which can be obtained either from plants (Ephedra
sp.), chemical synthesis involving resolution of a racemic mixture, or by
biotransformation of benzaldehyde using various yeasts. In the present review,
recent significant improvements in the microbial biotransformation are assessed
for both fed-batch and continuous processes using free and immobilised yeasts.
From previous fed-batch culture data, maximal levels of L-PAC of 10-12 gl-1 were
reported with yields of 55-60% theoretical based on benzaldehyde. However,
recently concentrations of more than 22 gl-1 have been obtained using a
wild-type strain of Candida utilis. This has been achieved through optimal
control of yeast metabolism (via microprocessor control of the respiratory
quotient, RQ) in order to enhance substrate pyruvate production and induce
pyruvate decarboxylase (PDC) activity. Processes involving purified PDC have
also been evaluated and it has been demonstrated that L-PAC levels up to 28 gl-1
can be obtained with yields of 90-95% theoretical based on the benzaldehyde
added. In the review the advantages and disadvantages of the various strategies
for the microbial and enzymatic production of L-PAC are compared. In view of the
increasing interest in microbial biotransformations, L- PAC production provides
an interesting example of enhancement through on-line control of a process
involving both toxic substrate (benzaldehyde) and end-product (L-PAC, benzyl
alcohol) inhibition.
Shin, H.S. and Rogers, P.L. (1996) Production of L-PAC from benzaldehyde using partially purified pyruvate decarboxylase (PDC). Biotechnol. Bioeng. 49, 52-62.
Shin, H.S. and Rogers, P.L. (1996) Kinetic evaluation of biotransformation of benzaldehyde to L-PAC by immobilized pyruvate decarboxylase. Biotechnol. Bioeng. 49, 429-436.
This one looks really good, a review article!
Oliver, A. L., Anderson, B. N. and Roddick, F. A. (1999) Factors affecting the production of L-phenylacetylcarbinol by yeast - a case study, Advances in Microbial Physiology, vol 41, pp1-41.
V.B. Shukla and P.R. Kulkarni, (2000) L-Phenylacetylcarbinol (LPAC) : biosynthesis and industrial applications,World J. Microb. Biotech. 16, 499-506 (UK).
Do Your Part To Win The War
(Newbee)
09-24-01 18:27
No 216691
SWIM translation:
OK all motovation inside starting with 4lbs white sugar and 2-3gallons h2o boiled and allowed to cool then added 1oz of active dry yeaqst (red and yellow jar)
Question is this canditas utilits ? what would work better? Is that super yeast for beer the right stuff?
About ten hours and then 60 ml distalled benzaldehyde was added.
4 days have elapsed and now I am thinking about what to do next?
Thanks
Hum did you get that?
(Distinctive Doe)
09-25-01 02:02
No 216825
Process parameters and reusability of the free cell mass of Torulaspora delbrueckii for the production of L-phenylacetylcarbinol (L-PAC).
AU: Shukla-V-B; Kulkarni-P-R {a}
SO: World-Journal-of-Microbiology-and-Biotec
April, 2001; 17 (3): 301-306..
PY: 2001
The effect of process parameters on the biotransformation of benzaldehyde to L-phenylacetylcarbinol (L-PAC) using a yeast isolate identified as Torulaspora delbrueckii was studied. The maximum yield of L-PAC obtained was (331 mg) per 100 ml biotransformation medium (glucose 3%, peptone 0.6% and at pH 4.5) from 600 mg of benzaldehyde with 8 h of reaction at 30+-2degreeC. Growing the organism in presence of 3% glucose reduced the biotransformation time to 120 min. Addition of 0.6% acetaldehyde (30-35%) lead to an increase in L-PAC yield to 450 mg%. Semi-continuous feeding of benzaldehyde (200 mg) and acetaldehyde (200 mul) four times at 30 min intervals could produce 683 mg of L-PAC/100 ml biotransformation medium. Chiral HPLC analysis of purified L-PAC and PAC-diol showed 99% enantiomeric purity. The cell mass was found to be reusable for biotransformation up to nine times when benzaldehyde and acetaldehyde levels were maintained at (350 mg and 350 mul)-(400 mg and 400 mul). At concentrations from 450 mg and 450 mul to 600 mg and 600 mul, however the cell mass could give efficient biotransformation only during one use.
Based on this you could get up to 100+ grams of L-PAC in a one day 5-gallon fermentation!!
Do Your Part To Win The War
(Distinctive Doe)
09-26-01 07:11
No 217273
I wonder what would happen if you put indolecarboxaldehyde in this fermentation??? Easy AMT?
Would a RP/I reaction reduce indole?
Do Your Part To Win The War
(Hive Addict)
10-12-01 01:07
No 223365
both yeast (Candida utilis) and pyruvate
decarboxylase (PDC)
What type of home brewing are these used for?
BISD: Built-in shit detector
(Hive Bee)
10-13-01 01:48
No 223832
pardon my stupidity, but:
this biosynth states that it creates l-ephedrine (after
reductive amination of l-pac).
However, upon reduction, won't this yield l-methamphetamine?
Forgive me if I am incorrect, but isn't the l-isomer of
methamphetamine much less effective, and is used in vicks'
inhalers?
Flames welcome.
Don't mind me. I'm mentally ill.
(Synaptic Cleft Mountaineer)
10-13-01 10:00
No 223952
l-ephedrine gives d-methamphetamine, and d-pseudoephedrine gives d-methamphetamine. Seems a little illogical at the first glance, but that is indeed the case.
(Hive Bee)
10-13-01 17:05
No 224026
yes, thanks rhodium, I realised that after I left and
was reading a doc on catalytic hydrogenation, and it
said (-)-ephedrine and (+)-psuedoephedrine yield
(+)-methamphetamine. I knew d-psuedoephedrine gave us
the right isomer, I assumed it was the same for ephedrine.
I've been looking around for a nice way to get some
benzaldehyde to use in this. looking at the
"phenylacetones.htm" file I found a patent reference
for the production of benzyl chloride by the chlorination
of toluene (it's the same thing as toluene with 1 xtra
Cl on the carbon outcrop).
The process bubbled Cl2 through Toluene, but it also said
that using "bleaching powder" at high temperature with
an acid provides nascent chlorine.
However, the guy who wrote this has a bitch about side
products. Now here is the NICE bit:
"I have discovered, however....,
by heating to a high temperature
an anhydrous mixture of toluene
and bleachin powder, with thorough
commingling, and without the use of
any acid, the chlorination of the
toluene is effected without any of
the disadvantages incident to the
processes above referred to. In
the absence of any added water the
chlorin from the bleaching powder
seems to go directly to produce
benzyl chlorid in the side chain
without attacking the nucleus, even
though the process is carried on in
the presence of iron, as in an
iron vessel"
Hmm. That looks nice. The reaction is pretty much:
Heat toluene to 90'C. Slowly add equal amount (weight)
of Ca(OCl)2 (pool shock), and raise temperature to
100'-105'C, leaving it there 1 hour or until reaction
complete. Settle. Decant of oil.
This oil contains 30-35% benzyl chloride and 65-70%
toluene.
However, We don't want benzyl chloride, do we? We want
BENZAL chloride. The patent goes on to describe that if
sufficient amount of Ca(OCl)2 is added, the toluene is
completely converted to Benzal Chloride and
Benzo Trichloride. Upon boiling in alkaline water, we
get Benzaldehyde and Benzoic acid. Voila.
Hope this will be a good addition to the home-brew-E
method, as many bees don't have that much benzaldehyde
lying around.
Check out the patent:
Don't mind me. I'm mentally ill.
(Hive Bee)
10-13-01 22:58
No 224105
Sorry you can't see the above. The patent is:
U.S. Pat.#1,280,612
Also, if the concentration of benzaldehyde in synthetic
almond essence is high enough, then the alcohol could
just be boiled off, and the rest plopped into the bucket.
Also, is it possible to get out the L-PAC in a fashion that
does not require vacumn distillation?
Ooooh! Here we go, info on the almond essence:
Bitter Almond oil is a light colourless liquid with a characteristic 'marzipan' scent. It main constituents are benzaldehyde (95%) and prussic acid (3%)."
bitchin! That's an extremely high amount. However, I'm not
convinced that this will be the concentration in OTC almond
essence. Perhaps if "imitation" essence is used, there will
be no prussic acid, as the essence will be made from
benzaldehyde, rather than via almonds. Maybe? Maybe not.
I'll be watching & working with you on this one.
Don't mind me. I'm mentally ill.
(Hive Bee)
10-13-01 23:06
No 224112
pardon my ever growing list of posts but...
It seems that the strain Candida Utilis is often
one of the organisms responsible for...
VAGINAL YEAST INFECTIONS!
This is really, really, really gross.
Okay, so who's going to be the first person to make
ephedrine from almond essence, vaginal yeast & sugar beets
and then meth from phosphorous from a human skull, and
iodine from stinky ol' kelp & dead fish.
Mmmmm. yummy. You can't get much more ORGANIC with this
organic chemistry.
This is bad. This is very very bad. Vaginal yeast. <Shudder>
Don't mind me. I'm mentally ill.
(Hive Bee)
10-14-01 00:41
No 224148
So ya's all want some Candida Utilis, ey?
There's pretty much three ways of getting it:
1. By accident.
2. From a culture collection - fork out $100
and get the third degree - "now ma'am, why did
you want to buy this sealed culture container?"
You should see the license agreement you have to
sign.
3. Go to a health store / pet store / big supermarket and
ask for Torula Yeast. After much searching around for this
damned strain of yeast, I found it.
Candida utilis (formerly Torulopsis utilis) is the yeast known as Torula Yeast. This yeast is also important in industry because it can utilize the pentose sugars from processed wood pulp used in making paper.
Gee, ya don't say?
Don't mind me. I'm mentally ill.
(Master Searcher)
10-14-01 01:29
No 224184
See US patent 2061136 for some other ways (other than MeNH2 and Al/Hg) of aminating L-PAC and 3,4- substituted L-PACs.
http://www.geocities.com/dritte123/PSPF.
(Hive Bee)
10-15-01 02:43
No 224546
SWIM's choice of action would be to aminate with
methylamine, then reduce with either palladium or
Urushibara Nickel (NiCl2 and Al).
A thought on the reductive amination of L-PAC:
the product of this is l-ephedrine, correct? If this
is so, and catalytic hydrogenation is applied (in leiu
of NaBH4 or LiAlH4), then shouldn't this l-ephedrine
be further reduced to d-methamphetamine at the same
time? (naturally, extra time/hydrogen would be given
to ensure complete reduction.
I assume that since methamphetamine has no more oxygen to
reduce, that there is no risk of over-reducing with this
method?
I am particularly interested in the above procedure, and
would love to hear some more from the author.
In wishing to simplify this procedure a bit more, mainly
for the sake of those who do not have vacumn distillation
apparatus, but also to reduce time and effort in create
large quantities, I am currently looking into different
ways of extracting the L-PAC from the fermentation mixture
that does not require the above-mentioned apparatus.
Perhaps a clever adaptation of selective solubility may
be in order? I'll check the library later, but if anybody
has solubility data regarding L-PAC, It would be very
welcome.
But then again, a fermentation is rather organic (gee ya
don't say), and much speculation could be made as to what
this mixture actually contains (anyone care for trying to
read a GC/MS?).
I'm sure people don't want to be snorting Torula yeast up
their nose - but then again, it is a diet suppliment, is
it no?
Tired but still working as always,
Encopo.
Don't mind me. I'm mentally ill.
(Stranger)
10-18-01 21:09
No 226338
Kudos to you for putting this info out there jacked. But I only have one last question left how do you perform a simple reductive amination. You see I'm a newbee and would like to get a simpler version of what I've seen posted thanks
________________________________________
Knowlege is power
(Hive Bee)
10-19-01 02:36
No 226514
Okie dokie, here's how it is.
Reductive Amination:
Well the "reductive" bit refers to reducing. Reducing is
synonymous with "hydrogenation". Let's say you have a
compound like the following:
# double bond
/ - \ single bond
O oxygen atom
C carbon atom
H hydrogen atom
O
#
H C H
\ / \ /
H-C C-H
| |
H-C C-H
/ \ / \
H C H
/ \
H H
Pardon the ascii art. This is cyclohexanone (keto form).
If we REDUCE this (HYDROGENATION) using catalytic
hydrogenation (that just means reducing with hydrogen and
a catalyst - something that speeds up the reaction, but
doesn't become part of the reaction products), we
will get:
OH
|
H C H
\ / \ /
H-C C-H
| |
H-C C-H
/ \ / \
H C H
/ \
H H
cyclohexanol.
Can you see how the hydrogen has "attached" itself to the
oxygen atom, by breaking one of the bonds that it had with
the carbon atom? If we continue to reduce we get:
H H
\ /
H C H
\ / \ / H
H-C C-H /
| | + O
H-C C-H \
/ \ / \ H
H C H
/ \
H H
cyclohexane
Can you see how further reducing(hydrogenation) has fully
removed the Oxygen atom from the compound? Now it has
bonded with another hydrogen atom, and no-longer has a
bond to the carbon atom, so it drops off as water (H2O).
Now the carbon is 2 bonds too short, and bonds with 2 more
hydrogens to "fill itself up".
This is how reducing works. If you look at ephedrine, you'll
see that it has an alpha-hydroxy group (another name for
an OH). If you reduce ephedrine, this O bonds with another
H atom and drops off as H2O. Then more H's come along to
"fill up the space", and we get methamphetamine (a powerful
and dangerous stimulant, a controlled substance in many
countries).
So now you know what REDUCING is. But you want to know what
reductive amination means?
Well, to AMINATE something refers to reacting a compound
with ammonia (NH3), or an ammonia-containing compound, such
as methylamine (NH2CH3) to produce an imine (never mind
what that means). Reductive amination is the action of:
1. reacting a compound with an ammonia-containing compound.
2. reducing the formed imine
Thus, when we reductively aminate safrole, we add NH2CH3,
and then we hydrogenate it (reduce it), to give us
3,4-methylenedioxymethamphetamine (pay honey).
If we're making honey from bromosafrole, we don't need to
reduce it, we just aminate it with ammonia (NH3). This is
why the bromosafrole method is popular with some people, as
it can be sealed up inside a container and sat for several
days (until it finished aminating), or put into a pipe-bomb
and heated to 130'C in hot oil for 2 hours (this aminates
faster).
In turning L-PAC into l-ephedrine, one could aminate with
methylamine in the pipe-bomb, and then reduce it with
sodium borohydride (NaBH4), or aluminum amalgam (Al/Hg),
catalytic hydrogenation with raney nickel, or possibly
even with an urushibara catalyst such as created from
nickel chloride (NiCl2) and aluminum. Many other ways
exist to reduce this formed imine. If one had the glassware,
the pipe-bomb would be replaced with a reflux condenser
and a round-bottomed flask, and the mixture of methylamine
in methanol and the L-PAC would be refluxed for the
necessary amount of time.
Many reductive aminations reduce and aminate at the same
time, such as the popular Al/Hg method (not actually the
Al/Hg method, but an extension a bee has made to it that
aminates and reduces at the same time).
So I hope that explains reductive amination. For more info,
visit Rhodium's site at ../rhodium/chemistry
and look at some of the info on different methods (of
interest to you might be the bromosafrole/halosafrole route
that uses the pipe-bomb for aminating. However, this
reaction aminates with ammonia, not methylamine as is
required for reductively aminating L-PAC).
Hope I could sort some things out for you.
And now I need a big dump (darned coffee).
Sphincterally yours,
Encopo.
Don't mind me. I'm mentally ill.
(Stranger)
10-19-01 17:08
No 226773
aha that was just the info that is going to be saving me some money dag nabbit. You my freind encopo are a lifesaver. I just read a post on microwave hydrogenation. Would you reccomend this method. No more stupid pills for me.-------------------------------------
(Newbee)
10-19-01 17:39
No 226786
What about reducing the imine with elektricity, is´nt that a way to go?
I still dont understand how one could sepate out l-pac out of the soup with acetone.
Is normal yeast for baking bread possible to use?
(Stranger)
10-19-01 17:50
No 226789
IN the halosafrole amination should I put the 1 mole of the L-PAC instead of the methylamine in the reaction to produce ephedrine
(Hive Bee)
10-19-01 21:45
No 226850
oooh, nononononono. That would have dire consqeuences.
Well, not dire, but not what you want. And wasteful
(two nice precursors down the drain).
I'm not sure what you want to do, but:
* If you want to use the halosafrole method to make
a halosafrole (such as bromosafrole), and then reductively
aminate it, then you should aminate it with AMMONIA, as
outlined nicely in the halosafrole/bromosafrole docs.
* If you want to make the L-PAC into ephedrine, then
you want to reductively aminate with methylamine, and
your choice of reducing procedure. A procedure that used
the pipe bomb FROM the halosafrole procedure to be able
to aminate the L-PAC with methylamine (NOT ammonia) in the
proper methanol solution, would aminate nicely, and then
you could procede to reduce.
As for extracting L-PAC with acetone, I'll be looking up
on the solubility of L-PAC in various solvents. Then I
will let you all in on the info. If someone can suggest
a better method than selective solubility, I'd be glad to
hear it (no, not distilling - many bees don't have/can't
afford the glassware).
And yes, I believe that a catalytic hydrogenation of some
description would be best for reducing the formed imine
(using electrolytically produced hydrogen, or if using
Urushibara, perhaps the nascent hydrogen generation from
aluminum would be sufficient (although I doubt it).).
Don't mind me. I'm mentally ill.
(Stranger)
10-20-01 06:24
No 227038
I was just reminiscing on my days of youth in my physical science class and remembered a procedure in which a large flashlight battery was used to seperate hydrogen and oxygen and as I remember it it produced hydrogen quite well. Hell the teacher even lit the test tube full of hydrogen when he was done. Nuff to convince me man.
(Hive Bee)
10-20-01 15:26
No 227143
In response to the use of normal baker's yeast in creating
L-PAC, I wouldn't suggest it. In just the same way that
cows are different animals to lions, you can't expect Simba
to be able to provide the solvent for your breakfast cereal,
no? Sure, SOMETHING will be created, and maybe even SOME
L-PAC will be created, but the conditions under which
baker's yeast proliferates would be different, and the end
products would not really be the ones we're after here.
On microwave hydrogenation:
Microwave hydrogenation seems to reduce reaction times when
compared to a reaction that provides hydrogen by a
dissolving metal such as aluminum, but not when compared to
using raney nickel in a parr-shaker hydrogenation setup.
Since Urushibara catalysts are far more OTC than raney
nickel, and don't have the pyrophoric properties, they would
be an easier choice for the chemical/apparatus deprived.
The use of one of the electrolytic hydrogenation device
mentioned throughout the hive (the ultrasonic refreshing
one looks VERY nice), would be good in this respect, if
a high-enough quality urushibara catalyst was prepared.
The hydrogen in this method is provided by the electrolysis
of water with an electrolyte (such as NaOH) dissolved in
it to provide H2 gas (and O2 gas), and this cuts down
on reaction time.
The use of ultrasonic refreshing acts to create the
environment at an atomic level of high-pressure and temp,
whilst being performed in an apparatus at room temp & press.
As someone said "the ultrasonic transducers literally
slam the hydrogen and ephedrine together". Of course, that
was in reference to reducing ephedrine to methamphetamine,
but a simliar principle would apply.
A note of warning though:
A method that myself and many other bees would find easy
may be more daunting and/or dangerous to an unexperienced
newbee.
The creation of Urushibara catalysts requires the
use of a carcinogenic and toxic compound - NiCl2 (nickel
chloride).
However, a chip a stray cat recently coughed up (this time
in my lounge) describes a person who has done some
experiments in creating the NiCl2 in situ. from nickel
oxide, and then directly proceeding with the creation of
an Urushibara catalyst.
I will post these findings later this week, after I have
deciphered the strange dialect that it is written in.
If this works, many newbees would be able to explore the
use of this exciting reducing agent, requiring only OTC
products, some time, and some caution.
By creating the NiCl2 in situ, one avoids cancerous tumours
forming in embarrasing places.
lymphatically bloatedly yours,
Encopo.
Don't mind me. I'm mentally ill.
(Hive Addict)
11-05-01 02:22
No 233119
€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€
*****AMERICAN JOURNAL OF GASTROENTEROLOGY*****
Nadir A Agrawal S King PD Marshall JB
Acute hepatitis associated with the use of a Chinese herbal product, ma-huang
[see comments]
In: Am J Gastroenterol (1996 Jul) 91(7):1436-8
ISSN: 0002-9270
Herbal medicines are widely perceived by the public as being healthful and
innocuous. A number of herbal medicines have now been linked with
hepatotoxicity. We report a case of acute hepatitis associated with the use of
ma-huang, a herbal product derived from plants of the Ephedra species, which is
advertised as being useful for causing weight loss and enhancing energy levels.
Given the lack of reports in the literature of hepatotoxicity with ma-huang and
ephedrine, we speculate that the ma-huang product our patient took contained
some other ingredient or contaminant or was misidentified. Our report and others
in the literature, which we review, indicate that the clinician should consider
herbal medicines as a possible cause of unexplained liver injury.
Registry Numbers:
299-42-3 (Ephedrine)
€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€
*****EMBO JOURNAL*****
Malek O Lattig K Hiesel R Brennicke A Knoop V
RNA editing in bryophytes and a molecular phylogeny of land plants.
In: EMBO J (1996 Mar 15) 15(6):1403-11
ISSN: 0261-4189
RNA editing has been observed to date in all groups of vascular plants, but not
in bryophytes. Its occurrence was therefore assumed to correlate with the
evolution of tracheophytes. To gain more insight into both the phylogeny of
early land plants and the evolution of mitochondrial RNA editing we have
investigated a number of vascular and non-vascular plant species. Contrary to
the belief that editing is absent from bryophytes, here we report mitochondrial
RNA editing in cox3 mRNA of the liverwort Pellia epiphylla, the mosses Tetraphis
pellucida and Ceratodon purpureus and the hornwort Anthroceros crispulus. RNA
editing in plants consequently predates the evolution of tracheophytes. Editing
is also found in the eusporangiate ferns Ophioglossum petiolatum and Angiopteris
palmiformis, the whisk fern Tmesipteris elongata and the gnetopsid Ephedra
gerardiana, but was not detected in Gnetum gnemon.cox3 mRNA of the lycopsid
Isoetes lacustris shows the highest frequency of RNA editing ever observed in a
plant, with 39% of all cytidine residues converted to uridines. The frequency of
RNA editing correlates with the genomic GC content rather than with the
phylogenetic position of a species. Phylogenetic trees derived from the slowly
evolving mitochondrial sequences find external support from the assessments of
classical systematics.
Registry Numbers:
EC 1.9.3.1 (Cytochrome-c Oxidase)
€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€€
*****ADVANCES IN BIOCHEMICAL ENGINEERING/BIOTECHNOLOGY*****
Rogers PL Shin HS Wang B
Biotransformation for L-ephedrine production.
In: Adv Biochem Eng Biotechnol (1997) 56:33-59
ISSN: 0724-6145
L-ephedrine is widely used in pharmaceutical preparations as a decongestant and
anti-asthmatic compound. One of the key intermediates in its production is
L-phenylacetylcarbinol (L-PAC) which can be obtained either from plants (Ephedra
sp.), chemical synthesis involving resolution of a racemic mixture, or by
biotransformation of benzaldehyde using various yeasts. In the present review,
recent significant improvements in the microbial biotransformation are assessed
for both fed-batch and continuous processes using free and immobilised yeasts.
From previous fed-batch culture data, maximal levels of L-PAC of 10-12 gl-1 were
reported with yields of 55-60% theoretical based on benzaldehyde. However,
recently concentrations of more than 22 gl-1 have been obtained using a
wild-type strain of Candida utilis. This has been achieved through optimal
control of yeast metabolism (via microprocessor control of the respiratory
quotient, RQ) in order to enhance substrate pyruvate production and induce
pyruvate decarboxylase (PDC) activity. Processes involving purified PDC have
also been evaluated and it has been demonstrated that L-PAC levels up to 28 gl-1
can be obtained with yields of 90-95% theoretical based on the benzaldehyde
added. In the review the advantages and disadvantages of the various strategies
for the microbial and enzymatic production of L-PAC are compared. In view of the
increasing interest in microbial biotransformations, L- PAC production provides
an interesting example of enhancement through on-line control of a process
involving both toxic substrate (benzaldehyde) and end-product (L-PAC, benzyl
alcohol) inhibition.
Registry Numbers:
EC 4.1.1.1 (Pyruvate Decarboxylase)
100-52-7 (benzaldehyde)
299-42-3 (Ephedrine)
67-64-1 (Acetone)
90-63-1 (1-hydroxy-1-phenyl-2-propanone)
palladium foil
(Distinctive Doe)
11-06-01 03:23
No 233432
Manufacture of L-phenylacetylcarbinol with Saccharomyces and preparation of L-ephedrine therefrom.
Horitsu, Hiroaki; Otsubo, Tetsuya. (Alps Yakuhin Kogyo K. K., Japan; Horitsu, Hiroaki).
Jpn. Kokai Tokkyo Koho (1997), 4 pp. CODEN: JKXXAF
JP 09234090 A2 19970909 Heisei.
Patent written in Japanese.
Abstract
L-Phenylacetylcarbinol (I) is manufd. from pyruvic acid (II) and PhCHO in a medium contg. C source with Saccharomyces yeast. L-Ephedrine (III) is prepd. from I manufd. by the above method. Bakers' yeast was precultured in a medium contg. molasses, salts, II, and vitamin B1 at 30° for 1 h. Subsequently PhCHO was added to the culture over 1 h and the incubation was continued for 5 h to give I. A mixt. of I, AcOBu, MeOH soln. of MeNH2, Adams Pt oxide, and MeOH was autoclaved at 3 atm H ro give III.
Method for isolation of L-ephedrine hydrochloride and sulfate salts.
Vondracek, Miloslav; Svoboda, Ivan. (Vyzkumny Ustav Antibiotik A Biotransformaci S.P., Czech Rep.). Czech Rep. (1996), 4 pp. CODEN: CZXXED
CZ 281218 B6 19960717 Patent written in Czech.
Abstract
L-Ephedrine hydrochloride and sulfate salts can be isolated from a reaction mixt. by reductive amination of D-(-)-1-phenyl-1-hydroxy-2-propanone at pH 10-13 and removing the catalyst with an org. phase, then bringing the pH to 1-6 with concd. HCl or H2SO4, with azeotropic distn. to remove 70-100% of the vol. of the water present and to release the desired salt.
L-Ephedrine from phenylacetyl carbinol.
Nebesky, Ferdinand; Souhrada, Josef; Jakl, Vladimir. (Czech.). Czech. (1978), 3 pp. CODEN: CZXXA9
CS 186027 19781130 Patent written in Czech.
Abstract
Phenylacetylcarbinol was hydrogenated at 50-5° and H pressure 202.6 kPa over a Pt catalyst in AcOBu contg. HCl with portionwise addn. of an aq. 35-40% MeNH2 soln. To give a mixt. of the L-(I) and DL-ephedrine, which was sepd. by crystn. to yield 88% I.HCl.
Do Your Part To Win The War
(Distinctive Doe)
11-06-01 03:29
No 233436
This is cool, If you are up to the plant cell culture, which is much more difficult than growing yeast.
Method for producing ephedrine from large-scale Ephedra cell culture.
Cha, Lihang; Jiao, Yuxia; Liu, Dalu; Zhu, Weixing; Zhang, Guozheng; Tang, Lianghong. (Institute of Chemical Metallurgy, Chinese Academy of Sciences, Peop. Rep. China; Xinjiang Tuofeng Pharmaceutic Industry Co., Ltd.). Faming Zhuanli Shenqing Gongkai Shuomingshu (2000), 13 pp. CODEN: CNXXEV
CN 1256316 A 20000614 Patent written in Chinese.
Abstract
Ephedrine is produced by inoculating Ephedra cell ZHJ-25CGMCCNo.0359 in a culture, culturing at 25° for 20-50 d by solid culturing, suspension culturing, or bioreactor culturing, and drying at 60°. The culture is prepd. by adding 0.7% agar to a soln. contg. KNO3 2.0-10.0, CuSO4 0.0001-0.001, NaH2PO4 0.3- 5.0, ZnSO4 0.001-0.01, MgSO4 0.5-3.5, (EDTA)Fe 0.1-2, Na3BO3 0.001-0.007, kinetin 0.001-0.01, (NH4)2SO4 0.5- 2.0, indoleacetic acid 0.001-0.01, CaCl2 0.5-5.0 mM, and sugar 1.0-10.0%, boiling, and sterilizing at 121° for 15 min. The culture may also contain phenylalanine 0.1-20 mM, fungus fermn. liquor 10-50%, vitamin B 0.4-0.5 mM, and nicotinic acid 0.078 mM.
Do Your Part To Win The War
(Distinctive Doe)
11-06-01 03:33
No 233440
Ephedra species: in vitro culture, micropropagation, and the production of ephedrine and other alkaloids.
O'dowd, N. A.; Mccauley, P. G.; Wilson, G.; Parnell, J. A. N.; Kavanagh, T. A. K.; Mcconnell, D. J. School of Botany, Trinity College, University of Dublin, Dublin, Ire.
Biotechnol. Agric. For. (1998), 41(Medicinal and Aromatic Plants X), 154-193.
CODEN: BAFOEG ISSN: 0934-943X. Journal; General Review written in English. CAN 128:101114
Abstract
A review with 102 refs.
This could bee very interesting.
Do Your Part To Win The War
(Distinctive Doe)
11-06-01 03:51
No 233448
1-Ephedrine.
Groeger, Detlef; Schmauder, Hans P.; Froemmel, Helmut. Ger. (East) (1966), 3 pp. CODEN: GEXXA8
DD 51651 19661125 Patent written in German.
Abstract
The title compd. (I) is prepd. by aerobic fermentation of a soln. of 18-25% beet molasses, 0.03-0.07% MgSO4.7H2O, 0.1-0.15% KH2PO4, 0.3-0.6% (NH4)2SO4 at pH 4.7-5.2, contg. small amts. of coenzymes, vitamin B, or whey and Saccharomyces cerevisiae, during 9 hrs. Within 4 hrs., 4 addns. of 0.2% C6H5CHO and AcH are added. A yield of 55-76% L-phenylacetylcarbinol is obtained which is treated by known methods to give I.
This one here looks like GOLD!!!
Factors affecting the production of L-phenylacetylcarbinol by yeast: a case study.
Oliver A L; Anderson B N; Roddick F A
ADVANCES IN MICROBIAL PHYSIOLOGY (1999), 41 1-45.
Ref: 112. Journal code: 2NT. ISSN:0065-2911.
Abstract
L-Phenylacetylcarbinol (L-PAC) is the precursor for L-ephedrine and D-pseudoephedrine, alkaloids possessing alpha- and beta-adrenergic activity. The most commonly used method for production of L-PAC is a biological method whereby the enzyme pyruvate decarboxylase (PDC) decarboxylates pyruvate and then condenses the product with added benzaldehyde. The process may be undertaken by either whole cells or purified PDC. If whole cells are used, the biomass may be grown and allowed to synthesize endogenous pyruvate, or the cells may be used as a catalyst only, with both pyruvate and benzaldehyde being added. Several yeast species have been investigated with regard to L-PAC-producing potential; the most commonly used organisms are strains of Saccharomyces cerevisiae and Candida utilis. It was found that initial high production rates did not necessarily result in the highest final yields. Researchers then examined ways of improving the productivity of the process. The substrate, benzaldehyde, and the product, L-PAC, as well as the by-products, were found to be toxic to the biomass. Methods examined to reduce toxicity include modification of benzaldehyde dosing regimes, immobilization of biomass or purified enzymes, modification of benzaldehyde solubility and the use of two-phase reaction systems. Various means of modifying metabolism to enhance enzyme activity, relevant metabolic pathways and yield have been examined. Methods investigated include the use of respiratory quotient to influence pyruvate production and induce fermentative activity, reduced aeration to increase PDC activity, and carbohydrate feeding to modify glycolytic enzyme activity. The effect of temperature on L-PAC yield has been examined to identify conditions which provide the optimal balance between L-PAC and benzyl alcohol production, and L-PAC inactivation. However, relatively little work has been undertaken on the effect of medium composition on L-PAC yield.
Do Your Part To Win The War
(Hive Addict)
11-06-01 06:05
No 233543
where did you find all that? I might be wrong but dont we want D-ephedrine
palladium foil
(Newbee)
11-06-01 07:08
No 233585
Here's two more possibilities for Benz from Toluene that I have found.
1) Photooxidation of Toluene in Cation-Exchange Zeolites
2) Oxygenation of Hydrocarbons using Nanostructured TiO2 as a Photocatalyst
Now number one is interesting, here is the puchline...
Toluene photooxidation was investigated in BaX and BaY (zeolites - BIG MAGIC). Toluene was introduced into the infrared cell at a pressure of approximately 10 torr and alllowed to equilibrate for one hour. Gas-phase toluene was subsiquently pumped out leaving strongly absorbed toluene. Quantitative measures of the toluene absorption indicated that the loading was approximately 2 toluene molecules per supercage. Molecular oxygen was then added to the infrared cell... sample was irradiated with broadband for 1hr... The end result is that BaY zeolite produced a 87% yeald then was nearly fully recycled.
Now BaY Zeolite can only be gotten from the petuitary of the invitro fetus of an extinct species of tree lizards, but otherwise the method is perfect.
The second synth is more viable however, needing only Tol, O2 (atm probably would work with patience) and TiO2 which is Titanium white used as a paint thickener and coloring...
Details were irritatingly thin, but Tol was oxidized in the presence of a TiO2 catalyst produced via flame deposition on a unnamed substrate. Two hours with bubbled O2 (rate not provided) under UV radiation from a Hg lamp with nearly theoretic conversion and selectivity.
Too tired to retype the reference tonight, but if people are interested I might tomorrow.
Let me know what you think, butI intend to try a TiO2 held in suspension by bubbling O2 thru Toluene. I figure recycle the O2 with a pump to really churn things up and hit it hard with UV from all sides via a reflector. Of course I just realized that: Toluene + O2 + Heat might lead to an exothermic side reactions involving the local authorities via kitchen rearangement which would reduce the selectivity of the outcome. Maybe I'll wait to hear from wiser bee's, and maybe some sleep, before I dream such things...
TrickE
And on the eight day, God created Meth...
... and hasn't done much of anything usefull since!
(Distinctive Doe)
11-06-01 18:18
No 233782
suk
No you want L-ephedrine.
I have a good book here
"Principles of Brewing Science" by George Fix
It has some pretty good basic info on all the chemical transformations in yeast. Mine is the old edition, I heard the new one was easier to understand, that could mean that he left out many good details but maybee he didn't.
Ok now to the goods.
Mr Fix says that Mg++ is a cofactor for the pyruvic acid decarboxylase enzyme. So I would say that you definately want/need some Magnesium in your media. Use epsom salt (MgSO4) in the amount recommended in one of the above media compositions.
Do Your Part To Win The War
(Stranger)
11-10-01 07:49
No 235102
Well I went to the grocery store to find me some Torula Yeast. No luck so's I decided to look for it on the net and guess what I found Ya'll
http://www.ohly.de/hutch.htm [url]http://
(Newbee)
02-17-02 20:05
No 270916
Since this same subject came up in the Crystal Meth forum, ressurecing this seems worthwhile.
My questions on the matter are:
Since both the precursor and the product (along with the side reaction products) are toxic to the biomass, is there perhaps some way to periodically rinse the yeast so as to refresh them and keep them viable?
Perhaps some sort of filtering to remove the nascent liquor for processing out the ephedrine, then returning the yeast to the fermenttion vessle to continue eating, reproducing and makeing more gold?
This one really seems like a good way to keep a low profile whilst continuing to cook.
(BTW, while vaginal yeas may be a strain of Candida, it is C albicans, not C. utilis)
burn-out only occurs if you go too fast in the wrong direction
(Newbee)
03-10-02 13:27
No 280073
Yeast & L-PAC
Look up:
WO 90/04631
US 6,271,008
WO 01/44486
US 5,173,413
DE 518029 (german)
GB 365535
all at:
http://ch.espacenet.com/espacenet/ch/en/
more to follow, if someone cares
ORGY
(who seeks for a bee experienced in catalytic hydrogenation/red. alkylation for putting together parts fitting so easily. Has done research on yeast not only in theory and now... PM!)
"I hope I'm becoming more eccentric. More room, you know.
More room in the brain."
(Newbee)
03-11-02 18:49
No 280685
Sorry in thy above post was a typo:
Thy german patent is:
DE1518029
it is identical with
GB1118293
whoooo not one noticed it.
seems me and my ketoalcohol will stay thy lonesome riders...
...no! Not all alone.
Shop closed.
ORGY
"I hope I'm becoming more eccentric. More room, you know.
More room in the brain."
(Newbee)
03-15-02 14:28
No 282983
Yes we proudly present, thy neverbefore shown image of thy mysterious
EPHEDRINE BIOSYNTH
A Pic, a pic, a pic!
As nobody wants to read patents nowadays (P-SAM ya thy guilty one ya did it intentional confess!
)Hey look, thats what it looks like, hold it before it undergoes racemisation! Oh, too late, its only PAC, no L-PAC anymore.
For those searching for valid information before starting thy part of free phantasting this may be helpful.
WO 90/04631
enough is enough
ya´ll get no colored one.
ORGY
"I hope I'm becoming more eccentric. More room, you know.
More room in the brain."
(Stranger)
03-21-02 16:52
No 285901
I read the posting and I have also tried to understand the reading on Biotransformation for L-ephedrine Production. by P.L. Rogers,H.S. Shin and B. Wang found in the pub lished by Springer on volume 56 Advances in Biochemical Engineering Biotechnology
First in the use of Candida Utilis, the max growth occured at PH 7 while the RQ( respiratory quotient ) value = 4 this value being the O2 AND CO2 concentration and the relative stirring of the metabolic step of the production.
While the article is good the data ca be confusing , also they use equipment out of reach for many of us.
However I would like to post the whole article for the interpretation of the reader , since many may not have access to the original work.
I would like to know if this would be cool or does it go against some type of policy of the forum ? The Journal that contains the article is pricy $150 dol. but its info is worth it.
(Hive Bee)
03-21-02 22:06
No 286014
Dont know if its cool or not,but I shure would like to catch a glimpse of that.Ive searched briefly on the net for it but only keep coming up with the same few paragraphs,telling me that it can be done and the basic procedure,not going into any specific detail.
Any info or links would be much appreciated by me.
Im pretty sure it would be cool to post some of it at least.
Its not on rhodiums page yet is it?
I realy think,seeing as the C.M board seems to have dried up when it comes to novel or new ideas and how the OTC sources of feedstock are becoming more of a bitch every day,that this subject should be the focus of all the meth bees.
Im sure within a couple of months we could get it cracked and form a relatively OCT method for biosynthimg e.
Then all we would have to worry about is when big brother makes yeast available by presription and starts regulating sugar purchases.
Whadda ya say bees?This would be one huge battle won in this war of ours.
BIOBALLZ
Then we just gotta get flinger to let us in on how to grow his meth plants and were laughing.
(Stranger)
03-23-02 02:28
No 286649
I wasn't able to post the whole refrence to the P.L.Rogers- H.S. Shin, B. Wang......BIOTRANSFORMATION FOR L- EPHEDRINE
so I sent it to the forum moderator see where he puts it, I also sent it to Rhodium maybe they will post it in its
entirety..................Question , is it possible to use fructose sugar as the source ? or just raw sugar cane sugar?
Here is recent finding , I thought it was interesting:
1: Biotechnol Bioeng 2001 Aug 20;74(4):317-25
Continuous production of (R)-phenylacetylcarbinol in an enzyme-membrane reactor
using a potent mutant of pyruvate decarboxylase from Zymomonas mobilis.
Goetz G, Iwan P, Hauer B, Breuer M, Pohl M.
Institut fur Enzymtechnologie der Universitat Dusseldorf im Forschungszentrum
Julich, D-52426 Julich, Germany.
The optimization of a continuous enzymatic reaction yielding
(R)-phenylacetylcarbinol (PAC), an intermediate of the L-ephedrine synthesis, is
presented. We compare the suitability of three pyruvate decarboxylases (PDC),
PDC from Saccharomyces cerevisiae, PDC from Zymomonas mobilis, and a potent
mutant of the latter, PDCW392M, with respect to their application in the
biotransformation using acetaldehyde and benzaldehyde as substrates. Among
these, the mutant enzyme was the most active and most stable one. The reaction
conditions of the carboligation reaction were investigated by determining
initial rate velocities with varying substrate concentrations of both aldehydes.
From the resulting data a kinetic model was inferred which fits the experimental
data with sufficient reliability to deduce the optimal concentrations of both
substrates for the enzymatic process. The results demonstrate that the
carboligation is most efficiently performed using a continuous reaction system
and feeding both aldehydes in equimolar concentration. Initial studies using a
continuously operated enzyme-membrane reactor gave (R)-PAC with a space-time
yield of 81 g L(-1). d(-1) using a substrate concentration of 50 mM of both
aldehydes. The yield was easily increased by cascadation of enzyme-membrane
reactors. The new strategy allows the synthesis of (R)-PAC from cheap substrates
in an aqueous reaction system. It thereby overcomes the limitation of by-product
formation that severely limits the current fermentative process.
PMID: 11410856 [PubMed - indexed for MEDLINE]
most recent article found and lots of research stuff on biotransformation at:
http://www3.ncbi.nlm.nih.gov/htbin-post/
......good reading
(Chief Bee)
03-23-02 03:02
No 286662
Sent it to me? Resend it please - rhodium@ziplip.com
(Hive Bee)
03-23-02 05:32
No 286705
what thy heck frucose?
I cannot say it will not work, but never heard about.
what thy yeast needs: vitamines, minerals its a lifeform.
happa, happa for yeast:
starting fermentation broth: molasses + yeast extract(does RECYCLING say anything to ya?) + glucose aka dextrose aka fucking plain sugar.
+ ceremony of sacrifice at choice.
it´s all in thy patents
hungry yeast is bad yeast
ORGY
"I hope I'm becoming more eccentric. More room, you know.
More room in the brain."
(Chief Bee)
03-23-02 10:57
No 286795
Here is the document I got from Java: ../rhodium/archive/biotransf
(Newbee)
04-27-02 07:02
No 301971
It is unlikely you could use Fructose. It may depend on the strain of yeast being used, but in brewing of drinkable stuff Fructose is added to sweeten the drink as it is unfermentable by the std yeasts used. Tastes twice as sweet as sugar, and one hell of a lot better than the artificial gunk.
When sugar is added, the yeast produces an enzyme which breaks sugar (sucrose) into Fructose and Glucose. The Glucose is then fermented to alcohol.
HTH, MMM
When the day is bad,and life's a curse
CHEER UP!!! Tomorrow may be Worse!!
('HAGAR' Comic)
(Hive Bee)
04-29-02 15:24
No 302974
MMM you are absolutely right.
Glucose = Dextrose but not Sucrose.
My thought was, that plain sugar can be supplied to feed the yeast, but as Dextrose is cheap and available this may be taken.
If betacyclodextrin is added it is possible to apply more benzaldehyde and better yields are reached. The adding of about 10% ethylalcohol may be substituted for betacyclodextrin.
thanks MMM
ORGY
(Hive Bee / Eraser)
07-29-02 20:11
No 338609
any new updates or breaks on this subject? jetson thought this had died long ago but to his surprise the last reply was just last april.
jetson's going to do some independant studying and will post anything new he finds but most likely it'll be more catching up than anything. the devil is so lonely
(Hive Bee)
09-21-02 03:14
No 358648
have a look at this quite new abstract and hope this sophisticated techniques will soon serve the clandestine chemist in his daily fight.
© Springer-Verlag 2002
Original Paper
Enzymatic (R)-phenylacetylcarbinol production in benzaldehyde emulsions
B. Rosche1, , N. Leksawasdi1, V. Sandford1, M. Breuer2, B. Hauer2 and P. Rogers1
(1)
School of Biotechnology and Biomolecular Sciences, University of New South Wales, Sydney NSW 2052, Australia
(2)
BASF-AG, Hauptlaboratorium, 67056 Ludwigshafen, Germany
Abstract. (R)-Phenylacetylcarbinol [(R)-PAC)] is the chiral precursor for the production of the pharmaceuticals ephedrine and pseudoephedrine. Reaction conditions were improved to achieve increased (R)-PAC levels in a simple batch biotransformation of benzaldehyde emulsions and pyruvate, using partially purified pyruvate decarboxylase (PDC) from the filamentous fungus Rhizopus javanicus NRRL 13161 as the catalyst. Lowering the temperature from 23°C to 6°C decreased initial rates but increased final (R)-PAC concentrations. Addition of ethanol, which increases benzaldehyde solubility, was not beneficial for (R)-PAC production. It was established that proton uptake during biotransformation increases the pH above 7 thereby limiting (R)-PAC production. For small-scale studies, biotransformations were buffered with 2-2.5 M MOPS (initial pH 6.5). High concentrations of MOPS as well as some alcohols and KCl stabilised PDC. A balance between PDC and substrate concentrations was determined with regards to (R)-PAC production and yields on enzyme and substrates. R. javanicus PDC (7.4 U/ml) produced 50.6 g/l (337 mM) (R)-PAC in 29 h at 6°C with initial 400 mM benzaldehyde and 600 mM pyruvate. Molar yields on consumed benzaldehyde and pyruvate were 97% and 59%, respectively, with 17% pyruvate degraded and 24% converted into acetaldehyde and acetoin; 43% PDC activity remained, indicating reasonable enzyme stability at high substrate and product concentrations.
E-mail: B.Rosche@unsw.edu.au
Phone: +61-2-93853895
Fax: +61-2-93136710
some perhaps valuable information on L-PAC:
L-PAC, aka laevo-phenylacetylcarbinol, is also named:
1-hydroxy-1-phenyl-2-propanone
1-hydroxy-1-phenylpropanone
l-1-phenylpropan-1-ol-2-on (phenylpropanolon) -> old german
laevo-phenylpropanolone
1-phenyl-2-ketoalcohol-(1)
1-phenyl-propanol-1-one-2
R-PAC
I am sure the babylonians have still more names for this simple compound. If somebody finds another one - PM me.
Also is this molecule well known since 1921, and heavily used in pharmaceutic industry, but there is no entry at chemfinder or another public database I have access.
Please, somebody who has access to better resources could look up the properties (boiling point, soluble in...)? Post them here or/and PM me?
THANKS!
Racemisation in basic solution is very fast, thats what I could dig up.
IMHO biotransformations are a sharp sword in the hand of clandestine chemistry. And I don´t speak of genetically engineered or otherwise dangerous organisms. All day everywhere yeasts are keeping a widely undeveloped (or not published) power of transformation. Enzyme extraction isn´t rocket science also.
I will be happy if I would get the L-PAC properties
ORGY
(Hive Bee)
09-29-02 04:17
No 361794
I found some more data for L-PAC:
Formula C9H10O2
Composition C, 71.98; H, 6.71; O, 21.31
Mol. Weight 150.1745
Exact Mass 150.06808
and the CAS NR is: 90-63-1
if now somebody with STN or BEILSTEIN access, AFAIK anyone living in GB and most people on universities have this for example, could be so kind and look this up and post the properties of L-PAC?
thanks
ORGY
<begin rant>
app. 30 seconds after a synth is posted they will come and show you all the goodies with the wise smile and this "we knew all this for long, boy" expression. Lot of Yogis here. Teacherism.
<end rant>
(Moderator)
09-29-02 23:31
No 362049
1-hydroxy-1-phenylacetone, aka Acetylphenylcarbinol, boils at 205-7° C at atmospheric pressure, 135-7° C at 24 mm Hg, 140-5° C at 11 mm Hg and 66° C at .2 mm Hg
It's miscible with most organic solvents, and more information can be found in JCS 1232 (1930), Ann 484 1 (1930), Compt Rend 198 1998 (1934), Biochem Z 245 238 (1932) & 230 320 (1931) and Bull Soc Chim 43 573 (1928) Phenylacetylcarbinol, aka 1-hydroxy-3-phenylacetone has a melting point of 48° C and boils at 144-5° C at 12-13 mm Hg, and more information can be found in Compt Rend 197 1328 & 1649 (1933)
(Hive Bee)
12-18-02 17:47
No 390532
Organikum Post No 388164 and there after.......that should have been here , and so it is now!.
(Stranger)
01-11-03 15:23
No 397227
Industrial Biotransformation:
http://www.ct.utwente.nl/ipp/docs/Papers
http://imb.usal.es/castellano/personales
roger2003
(not quite as irritable lately)
01-23-03 23:23
No 400937
I apologise if any of this has already been posted....
Here is the relevant bits from the SchmidetalEnzymes2002.pdf in the above post...
The flavoenzyme tryptophan 7-halogenase has been evaluated for selective chlorination of tryptophan and indole derivatives. The reaction is thought to proceed via the formation of an epoxide and ring-opening to a chlorohydrin followed by dehydration. Glaxo Wellcome Research and Development reported on the use of nucleoside oxidase. The enzyme was found to have a very broad substrate spectrum towards unnatural nucleosides.
Immobilization of the oxidase directly from crude extracts onto Eupergit-C resulted in stabilization of its activity, which also allowed reuse of the enzyme and an easy scale-up of the reaction. An interesting carbon–carbon bond formation reaction was reported for the preparation of (R)-phenylacetylcarbinol by carboligation of pyruvate and benzaldehyde using various pyruvate decarboxylases. Continuous production of (R)-phenylacetylcarbinol from acetaldehyde and benzaldehyde could be achieved using a mutant of pyruvate decarboxylase from Zymomonas mobilis in an enzyme membrane reactor (EMR) with space-time yields of 81g L–1 d–1.
Widespread application of enzymes in the chemical industry will depend on the ability to couple enzymatic and chemical steps. Chemoenzymatic reaction sequences profit from the high technical development level of both chemical and enzymatic reactions. DSM uses well-established amidase catalysis to produce enantiopure Cá-tetrasubstituted á-amino acids containing terminal double bonds that react to cyclic oligopeptides by Grubbs olefin metathesis. Lonza uses a sequence of nitrile hydratase catalysis, chemical hydrogenation, and amidase-catalyzed reactions to obtain enantiopure pipecolic and piperazine carboxylic acids from aromatic nitrile precursors. Making biocatalysis compatible with chemical multistep synthesis is one of the important future challenges for this new technology.
Conclusions
Today, applications of enzymes in the chemical industry are already well established and, given the current developments, the number of biocatalytic processes will continue to increase rapidly. The first hurdle, confidence in the new technology, has been taken and as more and more processes, technology and infrastructure are implemented, efforts in biocatalytic research and development will also increase. This will allow the discovery and application of new enzymes and biological counterparts for traditional chemical reactions and will facilitate the integration of enzymatic steps in chemical multistep syntheses.
Biotransformation processes for L-PAC production
Prof Peter Rogers
Biotransformation processes involving both yeast (Candida utilis) and pyruvate decarboxylase (PDC) are being evaluated for the production of L-phenylacetylcarbinol (L-PAC) from substrates benzaldehyde and pyruvate. L-PAC is an intermediate in the production of the decongestant and antiasthmatic pharmaceuticals, ephedrine and pseudoephedrine. Kinetic models for the process are under development together with a computer-based optimal substrate feeding profile for benzaldehyde.
Funding sources: Commercial (ICI 1994-6; other 1997-9)
Student involvement: One Postdoctoral Research Fellow and two PhD students, MAppSc and Honours students
Recent publications:
Shin, H.S. and Rogers, P.L. (1996) Production of L-PAC from benzaldehyde using partially purified pyruvate decarboxylase (PDC). Biotechnol. Bioeng. 49, 52-62.
Shin, H.S. and Rogers, P.L. (1996) Kinetic evaluation of biotransformation of benzaldehyde to L-PAC by immobilized pyruvate decarboxylase. Biotechnol. Bioeng. 49, 429-436.
Rogers, P.L., Shin, H.S. and Wang, B. (1997) Biotransformation for L-ephredrine production. Adv. Biochem. Eng. 56, 33-60.
Liew, M.K.H., Fane, A.G. and Rogers, P.L. (1997) Fouling effects of yeast culture with antifoam agents on microfilters. Biotechnol. Bioeng. 53, 10-16.
Liew, M.K.H., Fane, A.G. and Rogers, P.L. (1997) Fouling of microfiltration membranes by broth-free antifoam agents. Biotechnol. Bioeng. 56, 89-98.
http://www.biotech.unsw.edu.au/research2
http://www.iupac.org/publications/ci/199
Here is the details from this one...
Environmental biotechnology
The November issue also contains selected proceedings of an International Conference on Environmental Biotechnology ‘96 held in Palmerston North, New Zealand, 1–4 September 1996. The published conference papers offer a wide-ranging analysis of the potential of biotechnology to waste treatment, specific perspectives on environmental damage and remediation, industrial research on pollutant mitigation, research into the area of upflow anaerobic sludge blanket reactors, the biological treatment of food industry wastes and two examples of cleaner technology for developments from the pharmaceutical and paper industries. The cleaner technology examples included production of phenylacetylcarbinol (PAC) and production of phenylacetylcarbinol by yeast through productivity improvements and waste minimisation. LPhenylacetylcarbinol is a precursor for the synthesis of L-ephedrine and D-pseudoephedrine, two pharmaceuticals with nasal decongestant properties. LPhenylacetylcarbinol is generated biologically through the pyruvate decarboxylase-mediated condensation of added benzaldehyde with acetaldehyde generated metabolically from feed stock sugars via pyruvate. Some of the added benzaldehyde is converted through the action of alcohol dehydrogenase(s) to benzyl alcohol, an undesired by-product.
L-Phenylacetylcarbinol extracted from the fermentation broth is converted chemically by hydroamination in the presence of methylamine and hydrogen to L-ephedrine, and then by isomerization to D-pseudoephedrine. Bruce Anderson and colleagues at the Royal Melbourne Institute of Technology present a dual approach strategy to enhance the ratio of product to by-product generated and to minimize the waste treatment burden of the spent fermentation broth. He explains that benzaldehyde delivery to the fermentation has been modified to ensure that sufficient raw material is available, together with pyruvate, during peak periods of pyruvate decarboxylase activity, and that benzaldehyde is less available during periods of high alcohol dehydrogenase activity. The inorganic content of the spent fermentation broth has been reduced substantially by the partial substitution of raw sugar for molasses in the medium, with a reduction of molasses content by 60% resulting in an increase of phenylacetylcarbinol production.
Further work on the optimization of the concentration of carbohydrate, nitrogen and phosphate in the fermentation has been conducted and has led, he claims, to further productivity increases, together with reduced waste generation, resulting in an L-phenylacetylcarbinol process which is considerably ‘cleaner’ than the parent process.
Taken from some guy's resume I found in a search engine...
Head, Food & Fermentation Technology Division.
The project dealt with the biotransformation of benzaldehyde to L- phenyl acetyl carbinol (synthon for various drugs) using yeast isolate. Various aspects like standardization of the method of analysis, purification and identification of product & byproducts of the biotransformation using various techniques were standardised. Using yeast isolate, various process parameters for increasing yield of the product and reusability of the biocatalyst were studied. A novel immobilization method for the aforesaid biotransformation was standardised and the process parameters using immobilized cell system studied. The mass transfer coefficient, power consumption and hold up in a stirred tank reactor with a dual impeller system were studied for the growth and biotransformation medium and were compared with those for air-water system.
After establishing the correlation between these operating parameters, a scaling up of this biotransformation to 5 L was achieved in a systematic manner. Work was also carried out on the synthesis of various chiral compounds using a combination of chemical synthesis and biotransformation using Rhizopus arrhizus.
to be continued.
Intelligence is not the recall of knowledge, but the ability to use it. (Rainman was a retard)
(not quite as irritable lately)
01-23-03 23:40
No 400943
From
http://www.swsbm.com/Abstracts/Ephedra-A
*****ADVANCES IN BIOCHEMICAL ENGINEERING/BIOTECHNOLOGY*****
Rogers PL Shin HS Wang B
Biotransformation for L-ephedrine production.
In: Adv Biochem Eng Biotechnol (1997) 56:33-59
ISSN: 0724-6145
L-ephedrine is widely used in pharmaceutical preparations as a decongestant and anti-asthmatic compound. One of the key intermediates in its production is L-phenylacetylcarbinol (L-PAC) which can be obtained either from plants (Ephedra sp.), chemical synthesis involving resolution of a racemic mixture, or by biotransformation of benzaldehyde using various yeasts. In the present review, recent significant improvements in the microbial biotransformation are assessed for both fed-batch and continuous processes using free and immobilised yeasts. From previous fed-batch culture data, maximal levels of L-PAC of 10-12 gl-1 were reported with yields of 55-60% theoretical based on benzaldehyde. However, recently concentrations of more than 22 gl-1 have been obtained using a wild-type strain of Candida utilis. This has been achieved through optimal control of yeast metabolism (via microprocessor control of the respiratory quotient, RQ) in order to enhance substrate pyruvate production and induce pyruvate decarboxylase (PDC) activity.
Processes involving purified PDC have also been evaluated and it has been demonstrated that L-PAC levels up to 28 gl-1 can be obtained with yields of 90-95% theoretical based on the benzaldehyde added. In the review the advantages and disadvantages of the various strategies for the microbial and enzymatic production of L-PAC are compared. In view of the increasing interest in microbial biotransformations, L- PAC production provides an interesting example of enhancement through on-line control of a process involving both toxic substrate (benzaldehyde) and end-product (L-PAC, benzyl alcohol) inhibition.
Registry Numbers:
EC 4.1.1.1 (Pyruvate Decarboxylase)
100-52-7 (benzaldehyde)
299-42-3 (Ephedrine)
67-64-1 (Acetone)
90-63-1 (1-hydroxy-1-phenyl-2-propanone)
Taken from http://www.ephedra.nu/engels/
Also found same info mirrored here...
http://www.r00t-access.org/texts/ephedri
First site is interesting site devoted completely to ephedrine.
Synthesis of ephedrine
======================
Actively fermenting yeast transforms benzaldehyde into L-phenylacetylcarbinol (L-PAC), which can be reductively aminated to afford ephedrine or phenypropa-nolamine (depending on the amine used)
----------------------------------------
Title: ISOLATION, ANALYSIS, AND SYNTHESIS OF EPHEDRINE AND ITS DERIVATIVES
Abstract: A review is given of methods for the isolation, quantitative determination, and modification of the ephedrine alkaloids, and advances in this field of natural compound chemistry.
Author: Gazaliev, A. N.; Zhurinov, M. Zh.; Fazylov, S. D.; Balitskii, S. N.
Reference: Chem.Nat.Compd.(Engl.Transl.), 25, 3, 1989, 261-271
----------------------------------------
Title: The biosynthesis of ephedrine
Abstract: It is shown by 13C nuclear magnetic resonance spectroscopy that the labelled C2 fragment of <2,3-13C2>pyruvic acid is transferred intact into the C-methyl group and the adjacent carbon atom of the Ephedra alkaloids, norephedrine, ephedrine, norpseudoephedrine, and pseudoephedrine, in growing plants of Ephedra gerardiana.This finding serves to identify pyruvate as the elusive precursor of the aliphatic C2 terminus of the skeleton of the alkaloids.In earlier experiments with 14C-labelled substrates, label from <3-14C>pyruvic acid was incorporated mainly, but not exclusively, into the C-methyl group of ephedrine, and label from <2-14C>pyruvate was incorporated similarly into the carbon atom adjacent to the C-methyl group.A C6-C1 unit related to benzaldehyde or benzoic acid has long been known to generate the benzylic fragment of the carbon skeleton of the Ephedra alkaloids.It is likely that the carbon skeleton of ephedrine is generated from pyruvate and either benzaldehyde or benzoic acid, by a reaction analogous to the formation of acetoin or diacetyl from pyruvate and acetaldehyde or acetic acid, respectively.Key words: biosynthesis of ephedrine, Ephedra alkaloids, 13C NMR spectra, ephedrine, biosynthesis of pyruvic acid, incorporation into ephedrine 13C NMR spectra.
Grue-Sorensen et al, Can.J.Chem., EN, 67, 1989, 998-1009
----------------------------------------
Stereoselectrive Syntheses of Ephedrine and Related 2-Aminoalcohols of High Optical Purity from Protected Cyanohydrins
Abstract: Ephedrine and related optically active b-aminoalcohols can be prepared by zinc borohydride reduction of aryl O-protected magnesium imines and aryl a-hydroxyimimes which in turn are readily available from optically active cyanohydrins.
Jackson et al, Tetrahedron Lett., 31, 10, 1990, 1447-1450
----------------------------------------
Title: SYNTHESIS OF EPHEDRINE (DIALKYL PHOSPHOROTHIOATE)S. CRYSTALLOGRAPHIC STRUCTURE OF EPHEDRINE (DIETHYL PHOSPHOROTHIOATE)
Abstract: The synthesis of ephedrine (dialkyl phosphorothioate)s has been effected.The method of their preparation and their properties are described.The IR and PMR spectra of ephedrine (dialkyl phosphorothioate)s and some results of an x-ray structural investigation of the spatial structure of ephedrine (diethylphosphorothioate) are given.
----------------------------------------
Methods for Biocatalyst Screening
[url]http://www.google.com/search?q=cach
Again, sorry if some is redundant, I tried to check everything back thru this thread, but it's a bit of a mission. It seems as though the limited amount of info on the net regarding this matter is mostly the same stuff.
Intelligence is not the recall of knowledge, but the ability to use it. (Rainman was a retard)
(Hive Addict)
01-24-03 09:44
No 401069
The most effective way to do this biotransformation is:
benzaldehyde + acetaldehyde + molasses + brewers wort + some salts
whereby:
benzaldehyde from toluene and/or benzylalcohol
acetaldehyde from ethylalcohol
Yields up to 70% on benzaldehyde. But who minds if less, as the benzaldehyde not converted to L-PAC gets converted to benzylalcohol which can be oxidized to benzaldehyde again quite easily. Without acetaldehyde yields are cut by half.
Java has linked to the post with the writeup, if some is interested in the details. To the writeup is to add that it is not necessary to distill the L-PAC as written, but the extract from the fermentation broth can be reductive animated as is. No strong vacuum pump necessary! Tested with Al/Hg reductive alkylation.
The method is unbeaten by now (except by pure separated enzymes, >90% but extreme expensive).
Great work Placebo! Quite comprehensive, real good work.
ORG
(not quite as irritable lately)
01-24-03 12:26
No 401090
I will be happy if I would get the L-PAC properties
Tell me about it, I looked for about 12 hours today!
Org, check if there is anything useful on that last link and save it, otherwise it will disappear from the cache and the .pdf has to be paid for.
Intelligence is not the recall of knowledge, but the ability to use it. (Rainman was a retard)
(Hive Addict)
01-24-03 15:24
No 401113
(Rated as: excellent)
(R)-(+)-Mandelic acid + MeLi -> (+)-acetylphenylmethanol (45%)
Tetrahedron Lett.; EN; 28; 50; 1987; 6313-6316.
(R)-(-)-1-hydroxy-1-phenyl-2-propanone, bp 72-74°C/0.2mmHg
Collect.Czech.Chem.Commun.; EN; 55; 8; 1990; 2046-2051.
Collect.Czech.Chem.Commun.; EN; 55; 11; 1990; 2685-2691.
1-phenyl-propane-1,2-dione -H2/Pt/Al2O3-> (S)-(+)-1-hydroxy-1-phenyl-2-propanone
J.Catal.; EN; 204; 2; 2001; 281 - 291.
Constitution
Justus Liebigs Ann. Chem.; 526; 1936; 143, 170.
Biochem.Z.; 127; 1922; 338.
Chem.Zentralbl.; GE; 111; II; 1940; 1860.
(RS)-1-Hydroxy-1-phenyl-aceton (racemic), mp 9.5-11°C (ether, pentane)
Can.J.Chem.; EN; 68; 11; 1990; 2060-2069.
(RS)-1-Hydroxy-1-phenyl-aceton:
Boiling Point mmHg Reference
66°C 0.2 1
236-238°C 760 2
143-145°C 31 3
130-132°C 20 4
124°C 14 2
129-131°C 14 5
129-130°C 13 6
106-108°C 7 7-8
104-106°C 6 9
Ref. 1 J.Amer.Chem.Soc.; 73; 1951; 4284.
Ref. 2 Justus Liebigs Ann. Chem.; 526; 1936; 143, 170.
Ref. 3 Biochem.Z.; 127; 1922; 338.
Ref. 4 Bull.Soc.Chim.Fr.; <4> 33; 1923; 770, 771;
C.R.Hebd.Seances Acad.Sci.; 176; 1923; 313.
Ref. 5 Zh.Obshch.Khim.; 27; 1957; 1622,1625;engl.Ausg.S.1694,1697.
Ref. 6 Mem.Inst.Sci.Ind.Res.Osaka Univ.; 6; 1948; 96, 98.
Ref. 7 Yakugaku Zasshi; 77; 1957; 851,853; Chem.Abstr.; 1958; 1949.
Ref. 8 Yakugaku Zasshi; 76; 1956; 1250, 1253; Chem.Abstr.; 1957; 4309.
Ref. 9 Zh.Obshch.Khim.; 21; 1951; 183,185;engl.Ausg.S.199,201.
(R)-1-Hydroxy-1-phenyl-aceton
Boiling Point mmHg Ref:
65-67°C 0.4 1
100-102°C 0.01 2
123-124°C 12 3
124-125°C 12 4
118-119°C 8 5-6
Ref. 1 J.Med.Chem.; EN; 7; 1964; 427-433.
Ref. 2 Collect.Czech.Chem.Commun.; EN; 37; 1972; 3897-3901.
Ref. 3 Chem.Zvesti; 12; 1958; 687; Chem.Abstr.; 1959; 11289.
Ref. 4 Biochem.Z.; 127; 1922; 133.
Ref. 5 Chemia anal.; 3; 1958; 573; Chem.Abstr.; 1959; 13841.
Ref. 6 Chem.Zvesti; 12; 1958; 17,19; Chem.Abstr.; 1958; 10768.
Thats on the properties. Plus what lugh posted.
some tidbits out of patents and articles:
- L-PAC racemizes fast under basic conditions and higher temperatures. So keeping fermentation temperatures low and ph as acidic as possible (4,3 - 4,7, whereby 4,3 is only possible with yeast harvested in acidic enviroment). On the other side this offers a perfect way to tune your gear after your personal preferences.
- Useful for extraction of L-PAC from the fermentation broth are most nonpolar solvents as: (diethyl-)ether, petrolether, DCM and ethyl acetate. A continous extraction is favorable at low temperatures one can also boil away some of the water as long not dealing with serious amounts to synthesize. Salting out helps a lot.
- The yeast may be washed and reused after the fermentation one or two times, then undergo autolysis to gain yeast extract useful as feed in following fermentations. The extract reduces the needed amounts of salts and brewers wort.
- The yeast to use is not critical. Bakers yeast from the supermarket. Harvesting this yeast under acidic conditions aka adding up to 3% sulfuric acid hardens the yeast and makes a ph as low as 4,3. possible (if not hardened ph 4,7 to 5,2).
what brings up this sudden interest again I wonder?
perhaps the wintertime
ORG
Placebo: 12 hours? I tried it 2 weeks and didn´t get anything some time ago...
(Hive Bee)
02-03-03 23:13
No 404101
I thought I bring in this piece of the puzzle to this thread since it seems to fit in in the quest to a source of starting material......the elusive Ephedrine Hcl,
Organikum fills in some more pieces to the puzzle, here...java
Post 400469 (Organikum: "L-PAC reductive alkylation", Novel Discourse)
(Newbee)
02-19-03 18:18
No 409782
Benzylic biooxidation of various toluenes to aldehydes by peroxidase
Russ, Rainer; Zelinski, Thomas; Anke, Timm
Tetrahedron Lett. 2002, 43: 5 791 – 794
A catalytic method is described for the oxidation of toluene and substituted derivatives to the corresponding benzaldehydes by hydrogen peroxide, using peroxidase. In most cases the respective benzoic acid was produced as a byproduct. The reaction proceeds under mild conditions in an aqueous medium.
(Hive Addict)
04-20-03 01:34
No 428138
Substituted benzaldehydes are often used as feedstock in industrial chemistry. The selective oxidation of aromatic methyl groups to the respective aldehyde is, however, difficult.[1] The chemical oxidation of the methyl group commonly proceeds directly to the carboxylic acid. We therefore chose to investigate enzymatic methods, because enzymes can be chemoselective. We started with the laccase/2,2-azino-bis(3-ethylbenzthiazol
The transformation of 21 different substituted methyl aromatics by hydrogen peroxide and Coprinus peroxidase was tested. Only three of the compounds tested, p-cymene (4-isopropyl-toluene), m-cresol, and p-cresol, were not at all transformed into the corresponding benzaldehydes. All other 18 compounds were transformed into the respective benzaldehydes whereby the efficiency of the reaction varied (Table 1). Suitable substituents comprised methyl, halogen, methoxy, and nitro groups. It seems that the position of the substituent was more important than its composition. Ortho or para positions of the substituent to the methyl group were preferred against meta, except for the nitrotoluenes. o-Nitrobenzaldehyde was obtained in a low yield, whereas m-nitrobenzaldehyde was formed with yields comparable to the p-isomer. In the case of o-nitrotoluene, an interaction of the intermediate methyl cation radical with the nitro group perhaps prevented the formation of the aldehyde. o-Nitrotoluene was the only substrate that produced the alcohol derivative. The cresols were found not suitable for catalytic conversion by Coprinus peroxidase and hydrogen peroxide, probably due to polymerization reactions as described for lignin peroxidase.[4] The preparation with o-cresol immediately turned yellow after addition of the enzyme. A weaker discoloration to yellow was observed for m- and p-cresol, as well as toluene, 3-chloro-, 4-methoxy-, and 4-fluorotoluene. p-Cymene was transformed into two compounds that were not identified. A molecular mass of 132 and 136 inferred that none of the compounds was either an aldehyde- or carboxyl-derivative of p-cymene.
In summary, the method presented here used Coprinus peroxidase and hydrogen peroxide to oxidize a variety of toluene derivatives to their corresponding aldehydes. Further studies are underway to characterize the enzyme responsible, as well as improving the reaction conditions to provide better yields.
References
1. W.J. Mijs and C.R.H.I. de Jonge. Organic syntheses by oxidation with metal compounds, Plenum, London (1986).
2. A. Potthast, T. Rosenau, C.-L. Chen and J.S. Gratzl. J. Org. Chem. 60 (1995), pp. 4320–4621.
3. E. Fritz-Langhals and B. Kunath. Tetrahedron Lett. 39 (1998), pp. 5955–5956. Abstract | PDF (107 K)
4. M. Tien and T.K. Kirk. Proc. Natl. Acad. Sci. USA 81 (1984), pp. 2280–2284.
5. F. van Rantwijk and R.A. Sheldon. Curr. Opin. Biotechnol. 11 (2000), pp. 554–564. SummaryPlus | Full Text + Links | PDF (234 K)
6. V.P. Miller, A. Tschirret-Guth and P.R. Ortiz de Montellano. Arch. Biochem. Biophys. 319 (1995), pp. 333–340. Abstract | PDF (735 K)
7. J. Geigert, D.J. Dalietos, S.L. Neidlman, T.D. Lee and J. Wadsworth. Biochem. Biophys. Res. Comm. 114 (1983), pp. 1104–1108. Abstract-MEDLINE
There is a graphical abstract and a table that I can put up if anyone is interested.
(Hive Bee)
07-10-03 12:27
No 446145
Benzaldehyde (0,010 g, 0,1 mmol), sodiumpyruvate (0,205g), baker`s yeast (0,415 g) and citrate buffer (0,415 ml, pH 6) was placed into a 15 ml stainless steel vessel. The vessel was pressurised to 2000 psi by pumping dried liquid carbon dioxide into the vessel and stirred for 24 h at 33° C.
The vessel was than cooled to RT an slowly degassed.
82% yield of Phenylacetylcarbinol
US Pat 2003/0077769 Apr. 24, 2003
Patent WO0144486
(Stoni's sexual toy)
07-10-03 18:00
No 446214
I love 10mg synths.
Let's scale this one up by a factor of 1000:
Benzaldehyde (10g), sodiumpyruvate (205g), baker`s yeast (415 g) and citrate buffer (415 ml, pH 6) was placed into a 15 l stainless steel vessel. The vessel was pressurised to 2000 psi (150 bar) by pumping dried liquid carbon dioxide into the vessel and stirred for 24 h at 33° C.
The vessel was than cooled to RT an slowly degassed.
(Note: not one word about workup of the glob of slime, how to keep it from foaming all over the place etc...)
82% yield of Phenylacetylcarbinol.
I'm not fat just horizontally disproportionate.
http://www.whatreallyhappened.com
(Hive Bee)
07-10-03 18:32
No 446222
The workup in the patent (example 1) was carried out by radial chromatography, but in the above theads are a lot of methods for the workup.
You think, you need a vessel about 15 l for a 10 g (benzaldehyde) synthesis ?
I think it is only a question of the pressue (CO2) and you can take a 3-4 l vessel for a 30 g (benzaldehyde) synthesis.
roger2003
(Stoni's sexual toy)
07-10-03 19:44
No 446239
Probably so. The CO2 should be supercritical at that pressure (I think), so it will also act as a solvent. It's not the pressure that counts, some volume is definitely needed.
I'm not fat just horizontally disproportionate.
http://www.whatreallyhappened.com
(Stranger)
10-09-03 08:10
No 463523
So, has anyone tried using the Sodium Borohydride reduction with methylamine? Works with amazing yield for MDMA so is this a viable reduction method for L-PAC? I always thought this to be the best high yield, large scale amination I had seen. And let's face it, if one is going to go through all these hassles to produce a relatively valueless product such as ephedrine, when it can be acquired easier than one might think, it should be done on a large scale to justify the procedure. To me it does not seem much easier than the conversion of safrole to MDMA using a good wacker oxidation method yet it is worth far less.
Go Hard Or Go Home
(Stranger)
10-09-03 10:06
No 463553
I thought Id throw this patent up while we`re at it.
It might be of interest of those out there who are lazy like me.Sorry If it has been posted previously.
It describes a simular process as above,only it claims it can be worked in in non fermenting conditions,using common solvents at lower temps. Patent US6271008
Recycling may raise yield,but still, it looks a lot more like tweeer chemistry than ever before.
Sorry
(Hive Bee)
10-09-03 12:19
No 463560
Pyruvic acid [127-17-3] , 2-oxopropanoic acid, pyroracemic acid, a-ketopropionic acid, H3C–CO–COOH, Mr 88.06, is the most important a-oxocarboxylic acid. It plays a central role in energy metabolism in living organisms [8]. During exertion, pyruvic acid is formed from glycogen in the muscle and reduced to lactic acid [79-33-4]. In the liver, pyruvic acid can be converted into alanine [56-41-7] by reductive amination. Pyruvic acid was discovered and first described in 1835 by BERZELIUS [9].
Pyruvic acid is totally miscible with water, ethanol, and ether. Pyruvic acid exists in the keto form; the enol form cannot be detected [10].
Chemical Properties Pyruvic acid reacts as both an acid and a ketone. It forms, for example, oximes, hydrazones, and salts. 4,5-Dioxo-2-methyltetrahydrofuran-2-carb
On standing in aqueous solution, pyruvic acid polymerizes to higher molecular mass products via the dimeric ketoglutaric acid [19071-44-4] and the trimeric aldol product [12] , [13].
Like all 2-oxo acids, pyruvic acid eliminates carbon monoxide on treatment with concentrated sulfuric acid [14].
Oxidation of pyruvic acid gives acetic or oxalic acid [144-62-7] and carbon dioxide, depending on the conditions [15]. Lactic acid is obtained by reaction with reducing agents [1].
Reaction of a-amino acids with pyruvic acid gives, besides carbon dioxide, alanine [56-41-7] (transamination reaction) and the corresponding aldehyde with one carbon atom less [16]. Alanine is also obtained by reductive amination of pyruvic acid [1]. Phenylethylamines react with pyruvic acid to form the corresponding tetrahydroisoquinolines via the Bischler – Napieralski reaction [17]. Reaction with o-phenylenediamines gives quinoxalinols [18]. In a similar reaction the corresponding hydroxypteridines are obtained from 4,5-diaminopyrimidines and pyruvic acid [19]. Pyruvic acid reacts with aldehydes to form the corresponding a-keto-g-hydroxy acids, which then cyclize to butyrolactone derivatives [1] . Friedel – Crafts type reactions of aromatic compounds with pyruvic acid yield diarylpropionic acids. These compounds have achieved a certain degree of importance because they provide a good route to 1,1-diarylethylenes by dehydration and decarbonylation [15] , [20].
Production On an industrial scale, pyruvic acid is produced by dehydration and decarboxylation of tartaric acid [87-69-4] [21]. In this process, pyruvic acid is distilled from a mixture of tartaric acid and potassium and sodium hydrogen sulfates at 220 °C. The crude acid obtained (ca. 60 %) is then distilled in vacuum. The reaction temperature can be lowered to 160 °C by adding ethylene glycol [107-21-1] [22]. Pyruvic acid can also be obtained by the gas-phase oxidation of lactic acid [23] , but this process has not been successful industrially. In contrast, microbial oxidation of D-lactic acid by a new process results in high yields [24]. Microbial oxidation of 1,2-propanediol [57-55-6] to pyruvic acid has also been described [25]. Another process describes the hydrolysis of 2,2-dihalopropionic acids to pyruvic acid [26]. A process for the oxidation of methylglyoxal [78-98-8] with halogens has recently been published [27].
Uses Pyruvic acid is used mainly as an intermediate in the synthesis of pharmaceuticals. It is also employed in the production of crop protection agents, polymers, cosmetics, and foods.
Storage and Quality Specifications Pyruvic acid is stored and transported in tightly closed polyethylene containers. It can be kept for only a limited period and must therefore be stored in refrigerated areas at a maximum of 10 °C. At higher temperature, explosion can occur through spontaneous self-condensation [28]. The concentration of the commercial product is determined acidimetrically and decreases by ca. 1 % per month during storage.
Toxicology Pyruvic acid has a corrosive effect and irritates the eyes, skin, and respiratory passages.
[1] A. J. L. Cooper, J. Z. Ginos, A. Meister, Chem. Rev. 83 (1983) 321.
[8] K. Schreiber: Die Brenztraubensäure und ihr Stoffwechsel, Editio Cantor, Aulendorf 1956.
[9] J. J. Berzelius, Ann. Phys. 36 (1835) 1.
[10] A. Schellenberger, K. Winter, Chem. Ber. 92 (1959) 793.
[11] L. Wolff, Justus Liebigs Ann. Chem. 317 (1901) 1.
[12] H. Goldfine, Biochim. Biophys. Acta 40 (1960) 557.
[13] A. Schellenberger, E. Podany, Chem. Ber. 91 (1958) 1781.
[14] A. Bistrzycki, B. v. Siemiradzki, Ber. Dtsch. Chem. Ges. 39 (1906) 58.
[15] S. Patai, S. Dayagi, J. Chem. Soc. 1958, 3058.
[16] R. M. Herbst, L. L. Engel, J. Biol. Chem. 107 (1934) 505.
[17] G. Hahn, A. Hansel, Ber. Dtsch. Chem. Ges. 71 (1938) 2163.
[18] O. Hinsberg, Justus Liebigs Ann. Chem. 237 (1887) 327.
[19] G. B. Elion, G. H. Hitchings, P. B. Russel, J. Am. Chem. Soc. 72 (1950) 78.
[20] Bayer, Patent DE2830953 , 1978 (W. Meyer, H. Rudolf, E. Cleur, E. Schoenhals). = Patent US4369206
[21] J. W. Howard, W. A. Fraser, Org. Synth. Coll. Vol. 1 , 475. http://www.orgsyn.org/orgsyn/prep.asp?pr
[22] J. D. Riedel, Patent DE281902 , 1913.
[23] C. H. Boehringer Sohn, Patent DE523190 1931 (F. Zumstein).
[24] BASF, Patent EP313850 1988 (B. Cooper).
[25] Y. Izumi, Y. Matsumura, Y. Tani, H. Yamada, Agric. Biol. Chem. 46 (1982) 2673.
[26] Dow Chemical, Patent US3524880 1966 (L. H. Lee, D. E. Ranck).
[27] BASF, Patent DE3219355 1982 (U. R. Samel, L. Hupfer).
[28] Sichere Chemiearbeit 29 (1977) 87.
(Moderator)
11-01-03 22:45
No 468182
While there are some changes necessary in the inputs and outputs for producing L-PAC, as opposed to simply growing yeast for protein; this article by Litchfield about culturing single cell proteins should bee very helpful in gaining understanding of what's required
Bacteria are also capable of growing on a variety of raw materials, ranging from carbohydrates such as starch, and sugars, to gaseous and liquid hydrocarbons such as methane and petroleum fractions, to petrochemicals such as methanol and ethanol. Suitable nitrogen sources for bacterial growth include ammonia, ammonium salts, urea, nitrates, and the organic nitrogen in wastes. A mineral nutrient supplement must be added to the bacterial culture medium to furnish nutrients that may not be present in natural waters in concentrations sufficient to support growth.
The bacterial species most likely to be used for singly-cell protein production grow best in slightly acid to neutral pH in the range 5 to 7. The bacteria should also be able to tolerate temperatures in the 35 to 45 C range, because heat is released during the bacterial growth. The use of temperature-tolerant strains will minimize the need for refrigerating the water that cools the fermentation vessel. Bacterial species cannot be used for single-cell protein production if they are pathogenic for plants, animals or humans.
Bacterial single-cell protein may be produced in conventional hatch systems in which all of the nutrients are supplied to the fermentor initially; the cells are harvested when they have consumed the nutrients and stopped growing. However, in the more advanced production methods the nutrients are supplied continuously in the concentrations needed to support bacterial growth and the cells are harvested continuously once the population reaches the desired concentration.
The concentration of the carbon and energy source usually ranges from 2 to 10 per cent in batch processes. In the continuous process the supply of the carbon source is regulated so that the concentration in the growth medium does not exceed that required by the growing bacterial cells. This concentration will generally be lower than those used in batch processes.
Maintaining sterile conditions during single-cell protein production is very important because contaminating microorganisms grow very well in the culture medium. The incoming air, the nutrient medium and the fermentation equipment must be sterilized in all bacterial single-cell protein processes, and sterile conditions; must be maintained throughout the production cycle.
In continuous processes the nutrients are replenished as they are consumed to maintain the concentrations needed by the bacteria. The solution containing the bacteria is drawn off, treated to cause the bacteria to agglomerate or flocculate, and centrifuged. The liquid may then be recycled in the fermentor while the bacteria are spray-dried and ground to yield the final product.
After the nutrients are sterilized and intoduced into a fermentation vessel and inoculated with the bacteria to be grown. The vessel, which is known as a 'bioreactor', must be supplied with sterile air and with cooling water to prevent the heat released during fermentation from building up and killing the cells. The cooling water is circulated in either the outer jacket of the fermentor or through internal cooling coils.
The vessels are also fitted with instruments that measure and control the pH and temperature of the contents and the concentration of dissolved oxygen. The exhaust air from the bioreactor contains carbon dioxide that may be separated and compressed for sale to industrial users of carbon dioxide.
After the bacteria are removed from the fermentation tank, they must he separated from the culture broth, which is usually done by adding chemicals that will cause the cells to clump and then centrifuging them. The separated cells are dried to yield a product that will be stable during shipment and storage. Finally, there must be equipment for grinding and packaging the cells and a system for treating and recycling the spent culture fluid.
Oxygen transfer to the cells in the fermentor is a critical factor in obtaining growth rates and yields that are economically satisfactory. A variety of fermentor designs can provide suitable aeration. The most commonly used are the baffled stirred-tank reactor and the air-lift fermentor.
Although considerable research was conducted on the production of bacterial single-cell protein during the 1960s and early 1970s, Imperial Chemical Industries (ICI) in the United Kingdom developed the only process to reach a commercial scale of operation. In the ICI process the bacterium Methylophilas methvlotrophus, which has a generation time of about 2 hours, is grown continuously with methanol as the substrate and additional nutrients including ammonia and the minerals phosphorus, calcium and potassium. The company developed for the process a unique air-lift fermentor with a capacity of 1500 cubic metres. The fermentor design minimizes the requirements for cooling the vessel and the problem of oxygen limitation.
Schematic diagram of a baffled stirred tank fermentor. The air introduced into the fermentor is dispersed by the propellor-like agitator. The baffles projecting from the side of the tank shown in cross-sectional view help to ensure that the contents of the tank are thoroughly mixed and oxygenated.
In 1980 ICI commissioned a plant, with the capacity of prodicing 50 000 metric tons of single-cell protein every year, at Billingham, England. The plant has since been operated intermittently, with a production of 6000 metric tons per month. The bacteria grow on methanol as their energy source. Two metric tons of methanol yield about 1 metric ton of dry 'Pruteen' single-cell protein. The dried product, which contains about 72 per cent protein and 8 per cent moisture, has been sold as an animal feed supplement in Western European markets.
With soybean meal now costing just $190 per metric ton, however, 'Pruteen' is no longer competitive as an animal feedstuff and the plant is not being operated on a commercial scale at present. Nevertheless the development of the ICI process for making the bacterial single-cell protein exemplifies the application of modern chemical engineering to the field of biotechnology.
During the development of 'Pruteen' ICI scientists investigated the possibility of improving the conversion of methanol it single-cell protein by genetically modifying the ability of M. methvlotrophus to use ammonia. They introduced into the bacteria a gene for an ammonia-assimilating enzyme that is more efficient than the endogenous bacterial enzyme. Although the new gene was stable in the bacteria and was expressed there, only a 3 to 5 per cent increase in single-cell protein yield was, obtained with the genetically modified strain of bacteria.
Yeasts
Modern technology for producing yeast single-cell protein has largely developed since World War II. Today, yeast products for human or animal consumption are produced on a commercial scale in many countries. In addition, baker's yeast, which is grown on molasses, is sold as a food flavouring and nutritive ingredient in addition to being used as a leavening agent.
Yeast can be grown on a number of substrates. These include carbohydrates, both of the complex type, such as starch, and of the simple type, such as the sugars glucose, sucrose and lactose. Alternatively, sugar-containing raw materials such as corn syrup, molasses and cheese whey can be used. Some yeasts are able to grow on straight-chain hydrocarbons, which are obtained from petroleum, or on ethanol or methanol.
In addition to a carbon source, a nitrogen source is required. The nitrogen can be provided by addition of ammonia or ammonium salts to the culture medium. A supplement of mineral nutrients is also required.
The requirements for production of yeast single-cell protein are similar to those previously described for production by bacteria. The yeast should have a generation time of about 2 to 3 hours. It should he pH- and temperature-tolerant and genetically stable, giving satisfactory yields from the substrate used, and not cause disease in plants, animals or humans.
The, technology for producing yeast single-cell protein is also similar to that for making the bacterial products. The baffled, stirred-tank fermentor is the most common type of vessel for yeast single-cell protein production, but air-lift fermentors are also used. As in the bacterial cultures, heat is released during yeast growth and the fermentor must be provided with a cooling system.
The yeast fermentations may be operated either in the batch or continuous mode, or by a third mode called 'fed-batch'. In fed-batch processes the substrate and other nutrients are added in an incremental manner to meet the growth requirements of the yeast while maintaining very low nutrient concentrations in the growth medium at any time. This method yields 3.5 to 4.5 per cent dry weight of product compared with the 1.0 to 1.5 dry weight of product yielded by batch cultivation. Cells grown by fed-batch processes are harvested as they are in the batch: mode of production.
Although batch and fed-batch culture systems have been used in baker's yeast production for many years, only recently has the technology been available for monitoring and adjusting the pH and substrate concentrations to permit the continuous type of operation. Yeast cell concentrations of up to 16 per cent (dry weight) can be obtained in the continuous culture operations.
Yeasts have certain advantages over bacteria for production of single-cell protein. For one, the yeasts tolerate a more acid environment, in the range of 3.5 to 4.5 instead of the near neutral pHs preferred by bacteria. Consequently, yeast processes can be operated in a clean but non-sterile mode at a pH of 4.0 to 4,5 because most bacterial contaminants will not grow well in this degree of acidity. For another, yeast cell diameters are about 0.0005 centimetres as compared with 0.0001 centimetres for bacteria. Because of their larger size yeasts may be separated from the growth medium by centrifugation, without the need for a flocculation step.
Yeast single-cell protein production depends upon meeting the oxygen demand of the growing cultures. Yeasts grown on carbohydrates generally require about 1 kilogram of oxygen per kilogram dry weight of cells and when grown on hydrocarbons they need about twice that much. Air, which is sterilized by filtration, is supplied to the fermentor through a perforated screen or perforated pipes in the bottom of the vessel, or by a rotating aeration wheel or air-lift device similar to those used for culturing bacterial cells.
Yeast single-cell protein may be produced either under sterile, or clean but non-sterile, conditions. In a typical batch or fed-batch non-sterile operation in which carbohydrate is used as the carbon and energy source the medium is sterilized by passing it through a heat-exchanger and then charged into clean fennentors. Contamination control is based on having a pH of 4.0 to 5.0, supplying sterile air, and maintaining large populations of yeast cells in the fermentor to overwhelm any small numbers of contaminating microorganisms. In some continuous yeast fermentations that use hydrocarbons or ethanol as the substrate, completely sterile conditions may he needed to achieve the desired yields and product quality.
Candsda units, which is known as torula yeast and used both as an animal feed supplement and for human consumption, is manufactured from a wide range of raw materials, among which are ethanol, the sulphite waste liquor from paper mills, normal paraffin hydrocarbons and cheese whey.
Chemistry is our Covalent Bond
(Horrible Personality)
11-25-03 22:12
No 473151
thats a good piece of information again, love it.
But the problem is actually that ppl are talking here about biosynthesis who are obviously not even able to make some acetal or benzaldehyde from OTC chems. Thats the only problem - the easy biosynth is posted already. Not more difficult as brewing beer - what is not so easy as some may think btw..
One last piece of information:
How to determine the biosynthesis process is actually over?
- it doesnt smell like oil of bitter almonds anymore.
thats it, the technique is called "nosing", ask DWARFER.....
I am outa this.
done
ORG
(Hive Addict)
11-28-03 00:53
No 473520
thats a good piece of information again, love it.
But the problem is actually that ppl are talking here about biosynthesis who are obviously not even able to make some acetal or benzaldehyde from OTC chems. Thats the only problem - the easy biosynth is posted already. Not more difficult as brewing beer - what is not so easy as some may think btw..
No, that's not true sorry. Senior chemistry bees have continually given the misleading impression that there was vital or atleast important information mission necessary to success.
What do you mean beer-making is hard?!?!?!?!!?!??! That's an infuriating implication to make at this late stage.
Oh so it is a walk in the park then, I thought so, thats what the first references I've read translated to a long time ago..... seemed straightforward and complete enough, it seems it always was.
I think Ill trust the references on the yeast species, candida utilis.
The benzaldehyde and acetaldehyde was always something to work at, it's just that uncertainties were portrayed.
Ohh you meant difficult as in you'd have to follow instructions and a few parametres.
You don't need 2 Bee High to follow the goings-on @The-Hive--Just Addikted!
(Horrible Personality)
11-28-03 14:00
No 473625
I didnt write it is difficult - but it is more than throwing some yeast in a bucket with sugar.
I see Java has provided the post number but no direct link. Here it is:
Post 388164 (Organikum: "substituted hydroxyphenylacetone", Novel Discourse) and followups
and
Post 400469 (Organikum: "L-PAC reductive alkylation", Novel Discourse)
The procedure as described is straight after an east german patent and works fine. Tried and true. And probably the way most ephedrine on the world is manufactured in China and India. Low tech process.
yeast:
actively fermenting saccaromyces cerevesiae aka bakers yeast is to use - actively fermenting says aerated and feeded with dextrose. ph-control is important. Brewers wort and acetaldehyde are important but can be substituted by diminished yields.
If your batch after 10 hours of active fermentation still smells like christmaspunch use another sort (strain) of bakers yeast.
Brewers wort here is wort before the hops are added - the temperature of the wort must never have exceeded 78°C or the essential enzymes are destroyed and its near to worthless.
and now I am realy out here.
ORG
(Chief Bee)
12-04-03 02:16
No 474575
Review: Application of beta-keto acid decarboxylases in biotransformations
H. Iding, P. Siegert, K. Mesch and M. Pohl
Biochimica et Biophysica Acta (BBA) - Protein Structure and Molecular Enzymology, 1385(2), 307-322 (1998) (../rhodium/pdf /l-pac.biotra
DOI:10.1016/S0167-4838(98)00076-4
Abstract
The advantages of using enzymes in the synthesis of organic compounds relate to their versatility, high reaction rates, and regio- and stereospecificity and the relatively mild reaction conditions involved. Stereospecificity is especially important in the synthesis of bioactive molecules, as only one of the enantiomeric forms usually manifests bioactivity, whereas the other is often toxic. Although enzymes which catalyze asymmetric carbon-carbon bond formation are of great importance in bioorganic chemistry, only a few examples are known for thiamin diphosphate (ThDP)-dependent enzymes, whereas transformations using e.g. aldolases, lipases and lyases are well documented already. The present review surveys recent work on the application of pyruvate decarboxylase and benzoylformate decarboxylase in organic synthesis. These enzymes catalyze the synthesis of chiral beta-hydroxy ketones which are versatile building blocks for organic and pharmaceutical chemistry. Besides the substrate spectra of both enzymes amino acid residues relevant for substrate specificity and enantioselectivity of pyruvate decarboxylase have been investigated by site-directed mutagenesis.
(Horrible Personality)
01-24-04 17:40
No 484386
Production of yeast-alginate pearls:
(hope alginat is alginate in english LOL)
2,5g yeast is mixed with 3ml waterby shaking.
0,25g sodiumalginate is shaken with 7ml of water.
Then the yeast suspension is given to the dissolved alginate.
0,3g CaCl2 is dissolved in 15ml water and stirred like hell (vortex stirring). Whilst stirring the yeast/alginate mixture is added to the CaCl2/water dropwise using a syringe. The syringe should be hold vertically when doing this.
3mm diameter pearls of yeast enclosed in alginate will drop out and can be removed by using an ultrahightech teasieve device.
voila - immobilized yeast.
I know I said I am out here, but thats to good to disclose it I believe. And it might come VERY handy not only in the L-PAC process which btw is called "acloin condensation" in newer publications. Ah these chemists - always a new name for an old reaction.....
ORG
(Hive Bee)
01-29-04 01:35
No 485174
sorry about that outburst up there, it looks disgraceful and immature now. what a **cken baby.
=============================
It seems like the biotranformation is highly dependent on Can high PDC activity the type of PDC and yeast one can get a hold of, including mutated forms.
Organikum, I'm not clear-minded regarding whether acetaldehyde addition is beneficial in a batch process...
Is alcohol dehydrogenase activity decreased with higher concentrations of acetaldehyde already present, necessarily?
Is it possible to produce acetaldehyde in situ before addition of benzaldehyde in a second phase of the fermentation process? Perhaps after the addition of more yeast, and developing this second culture.
(see: 'LPhenylacetylcarbinol is generated biologically through the pyruvate decarboxylase-mediated condensation of added benzaldehyde with acetaldehyde generated metabolically from feed stock sugars via pyruvate...')
Do we know the response time of increased PDC activity as a result of reduced aeration? (For the purpose of having high PDC activity at the moment of substrate addition(s))
Do we know the amount of yeast that is required for developing enough of it in a specific time for a given quantity of benzaldehyde feedstock? An estimate?
Tell me if I'm unclear, it's happened before.
(Horrible Personality)
01-29-04 17:16
No 485311
- Acetaldehyde works as a hydrogen acceptor - it is essential for good yields.
- Too much acetaldehyde as benzaldehyde kills the yeast.
- The yeast "cycles" through phases of high PDC activity and lower activity. The monitoring of these cycles is high tech and warrants not the efforts IMHO.
- Ethylalcohol may be added to suppress the benzaldehyde -> benzylalcohol transformation, about 10% to 15% is ok, more may disable or even kill the yeast.
- The benzaldehyde -> L-PAC transformation is depenedent on MANY factors not only PDC activity - yeasts with lots of PDC in a highly active state dont give the best results.
Do we know the amount of yeast that is required for developing enough of it in a specific time for a given quantity of benzaldehyde feedstock? An estimate?
Thats covered in my posts I linked to before.
The acetaldehyde added is NOT converted to L-PAC, only the acetaldehyde which is made in situ from the pyruvate is converted to L-PAC. This is not acetaldehyde in a pure form but a kind of nascent acetaldehyde compound.
It works without the addition of acetaldhyde with lowered yields.
The process which is used in the production of heaps of ephedrine to lowest prices in China and India is described in my former posts. No need to make it overly complicated.
Get yourself a nice b-dehyde synth and an acetaldehyde synth - thats the problem to solve. The biosynth is easy.
(Newbee)
02-07-04 18:28
No 487103
Does anyone have information concerning US patent #5,420,022
phenylacetic acid from yeast and phenylalanine. How much phenylalanine is added to the brew, and how does one extract the phenylacetic acid from the brew? Can nutrasweet be used as more of it can be dissolved in water,so it is easier to mix?
02-14-04 04:44
(Rated as: insignificant)
(Hive Bee)
03-06-04 16:32
No 493430
...well, that -OH is obsolete anyway.
Patent CA2414742: Microbial Production Of R-Phenylacetylcarbinol By Biotransformation Of Benzaldehyde By Filamentous Fungi
Abstract: Process for the production of R-phenylacetylcarbinol by biotransformation of benzaldehyde by filamentous fungi.
Just some investigations of R-PAC production by filamentous fungi selected from the group of Rhizopus, Neurospora, Polyporus, Fusarium, Monilia, Paecilomyces, Mucor.
“The real drug-problem is that we need more and better drugs.” – J. Ott
(Wonderful Personality)
03-08-04 13:18
No 493730
Its the same.
l -(laevo)- phenylacetylcarbinol is identical with R-Phenylacetylcarbinol.
So many compounds but much more names - oh Babylon, oh Babylon
.S we have the funny fact arising that:
l-PAC = L-PAC (whats wrong but often used anyways) = R-PAC
This doesnt matter anyways as "phneylacetylcarbinol is the wrong name also - a mistake made by the discoverers Neuberg and Hirsch.
Look above there are all names (known to me) listed.
(Hive Bee)
03-08-04 14:03
No 493737
Thanks for the correction Organikum. I already figured that out, but it was too late to edit the post. Yes, it is just a different nomenclature, but I always mix up those R,S/D,L nomenclatures if I don’t think enough about stereochemistry and I almost never think
.Well, sometimes I do think but the results of such an irresponsible activity might be wrong
. Did anybody ever considered the following transformation:
Similar mechanisms are already known for alpha-hydroxy-ketones, especially in the heterocyclic chemistry. Here we have one more driving force to push the equilibrium on the right: the conjugation of the enamine and latter of the carbonyl double bond with the benzene ring. One additional push would be to eliminate the water that forms during the enamine formation. Azeotropic distillation with a Dean-Stark trap would be a little hard to acomplish due to the volatility of dimethylamine, diethylamine, methylethylamine etc. N,N-dimethylcathinone is a relatively good stimulant, isn't it? And it is much more straightforward for the kitchen chemist than reduction to ephedrine and so on.
“The real drug-problem is that we need more and better drugs.” – J. Ott
(Wonderful Personality)
03-08-04 15:52
No 493748
You want it easy AND good?
Go:
benzaldehyde + bakers yeast = l-PAC
l-PAC (crude extract from the biosynth is ok) + NH3 = imine *
imine + Al/Hg = l-norephedrine
l-norephedrine + KOCN = carbamyl
carbamyl + HCl = "XYZ" surprise, surprise what can this bee....?
* hydroxyphenylacetones form stable imines, so the formation of the same in advance before reduction is possible and advantegous regarding yields which are bad if the reductive amination is done one-step as usual.
(Hive Bee)
03-08-04 17:07
No 493762
Very interesting book (5 MB)
Industrial Biotransformations:
A Collection of Processes by Andreas Liese (Author), Karsten Seelbach (Author), Christian Wandrey (Author)
http://www.amazon.com/exec/obidos/tg/det
roger2003
(Wonderful Personality)
03-09-04 01:07
No 493820
You wanted to say 150$$ I guess.
Thats a joke, there are hundreds of interesting books at amazon, you know?
Thousands probably.
weird.
03-09-04 10:16
(Rated as: insignificant)
(Hive Bee)
04-09-04 05:37
No 499839
I did a little research on pyruvic acid and discovered that it's right under our noses - well, when we're making our dinner it can make the eyes water... yes ONIONS! sorry if that was known to the reader... Pyruvic acid is produced during the decomposition of sugars (glucose i think) and it can reduce quite readily to lactic acid.
so all you need is a source of pyruvate decarboxylase, and with a few experiments one should find that a particular set of conditions is conducive to the PDC reacting with the pyruvic acid in such a way as to increase the chances of this decarboxylation occurring in proximity to the benzaldehyde so that one gets maximum transformation.
Or better still, is there a way to make benzaldehyde and pyruvic acid join together the right way without using an enzyme?
Pyruvic acid could probably be easily sublimation purified out of extract from onion - subzero alcohol extraction would extract solid pyruvic acid into the alcohol and then the material boiled off and the fraction boiling at 165 degrees distilled, and then sublimed onto a dry ice cooled dewar type condenser as it solidifies at 11.8°C.
This might be useful information for the biosynthesis process, as pyruvic acid probably doesn't have a very long lifespan in actively brewing warm vat... maybe it could be added with a dripping funnel thingy? I'm sure it wouldn't cost much to get a few litres of pyruvic acid from the supermarket...
(Hive Bee)
04-09-04 10:12
No 499867
Pyruvic acid could probably be easily sublimation purified out of extract from onion
Are you serious?
You can make pyruvic acid with the dry distillation of tartaric acid which is OTC and cheap (at least where I live). I don't remember the exact procedure but I'm sure it is as easy as it can get (maybe some sulphuric acid has to be added or something like that).
“The real drug-problem is that we need more and better drugs.” – J. Ott
(Hive Bee)
04-09-04 12:31
No 499877
the question is, would it help to add it? It is naturally formed as a part of the yeast's metabolism, (and virtually any organism that uses sugars to provide energy) but is the rate of production equal the rate of PDC production? If PDC was being produced in excess to the proportion of pyruvic acid, we could probably squeeze more out of it (?) If not, the whole line of thought is worthless... unless we can get PDC out of yeast somehow perhaps, or if it is commercially available. or... maybe there's some way to make pyruvic acid useful here somehow (chlorinate it and react with benzyl chloride perhaps?)
(just looking at molecules) oh i see decarboxylate tartaric acid one time and remove one hydrogen (or add an oxygen) and you get *tada* pyruvic acid.
I've been trying to nut out how exactly the PDC goes about joining these things together and what i've figured out is this: The pyruvic acid decarboxylates, and the free carbon bond remaining of what used to be pyruvic acid reacts with the aldehyde, which bonds by saturating the carbon with the acetaldehyde which is lacking a hydrogen (which is floating around from the decarboxylation), and the hydrogen goes onto the now single-bonded oxygen at the first carbon on the now phenylacetylcarbinol.
If one created reaction conditions of pyruvate decarboxylating in the presence of benzaldehyde, it doesn't make any difference how it is done so long as the process allows the freshly decarboxylated pyruvic acid take up the carbon bond. or am i missing some vital point here? The stereoselectivity is inherent in this reaction because of the way it reacts via the production of an aldehyde with an extra proton on the carbon with the aldehyde on it, because the keto group has some kind of interaction with the oxygen with a free electron. I think in this case we always want the OH and keto group to end up opposite each other, and i suspect that is exactly what they do. One could imagine that there would be some kind of interaction leading to a fairly predictable bonding formation.
Tricky thing is, how do we get the pyruvic acid to decarboxylate with benzaldehyde around? perhaps a high boiling ketone? pressure (*ducks flamethrower blasts*) ? *wince* ... ooh, maybe something simple like a natural organic ketone, eg carvone or something similar... That would dissolve the benzaldehyde well enough but the determining factor for the best ketone solvent/matrix/decarboxylation catalyst would be good solubility of pyruvic acid.
(Wonderful Personality)
04-09-04 13:14
No 499880
No. Pyruvic acid or molasses or even plain sugar makes no real difference. Molasses is even better for the nutrients it contains.
But pyruvic acid may be useful in another way.
- pyruvic acid and diluted H2SO4 gives acetaldehyde.
The biosynthesis is absolutely no problem. Acetaldehyde and benzaldehyde are the limiting factors here.
For the biosynth:
- permanent ph control and adjustment is essential.
(Hive Bee)
04-09-04 15:59
No 499900
- pyruvic acid and diluted H2SO4 gives acetaldehyde.
Any references ?
roger2003
(Newbee)
04-09-04 18:29
No 499907
bio brew tryptophan->tryptophol.
bio brew phenylalanine ->phenyl acetic acid.
now to do this can one use the same set up as is used for L-pac,I read that patent for phenyl acetic acid production but was wondering if it could be brewed more like beer as with the L-pac?
(Newbee)
04-09-04 22:09
No 499937
Here is a good Pyruvic acid patent.
Claimed 90% yield from cream of tartar and h2so4Patent US4136111
(Hive Bee)
04-10-04 06:23
No 499995
if only it made phenylacetic acid... no sorry, biobrew is gonna turn the material into phenylethanol or maybe phenylacetaldehyde (i'm fairly sure it's the alcohol)
phenylethanol is not going to be an easy way to start if one wants to end up with an amphetamine, since the amine has to go on there but it needs another methyl group after it as well...
(Newbee)
04-10-04 18:29
No 500077
phenylethanol can changed to phenyl acetic acid then ran through a tube furnace to get p2p right? what is needed to change phenylethanol to phenyl acetic acid? in patent 5420022 it seems they were producing phenyl acetic acid,the beer brewing set up would be somewhat different than their set up so maybe the the main product would be phenyl ethanol.so can I get more feed back on this, thank you.
(Hive Bee)
04-10-04 23:53
No 500117
phenylethanol probably oxidises to it's carboxylic acid equivalent via some means, ordinary ethanol certainly seems to. This requires some kind of bio-catalysing normally, i think, you can't just let ever clear sit open and expect it to oxidise, a lot of water is needed. Probably you just brew it up, then don't stop until the thing gets really acidic and full of acetic acid. I would think the same oxidation would occur, but then i can't be sure cos phenylethyl alcohol is a lot bigger.
If there's a way to make phenylacetic acid, then couldn't tryptophol also be turned into indoleacetic acid?
(Hive Bee)
04-18-04 09:56
No 501439
How about modifying this: Method for producing L-3,4-dihydroxyphenylalanine - European Patent (EP 0636695-A1) (http://l2.espacenet.com/espacenet/viewe
1: Instead of aiming for DOPA one could aim for alpha-methyl-DOPA by using catechol with alpha-methyl-serine. Possibly may not work but surely worth investigating.
People have discussed the conversion of alpha-methyl-DOPA to MDA via decarboxylation followed by methylenation but Rhodium told me that it wouldn't work. I can't see why not. But I suppose it's academic without any alpha-methyl-DOPA.
2nd point. Why not try the above Euro patent with hydroquinone and serine rather than catechol and serine? - to, possibly, get 2,5-dihydroxy-phenyl-alanine - which is just a decarboxylation and a methylation (I daren't go further) away from something interesting.
Apologies is this post interrupts the thread and is thought to be off-topic.
(Wonderful Personality)
04-24-04 21:45
No 502714
As acetaldehyde is very useful in the biosynthesis of l-PAC I want to point interested bees here:
Post 502712 (Organikum: "Rhodium wrote: Re: If you do not remove the...", Stimulants)
ferrous/ferric sulfate + hydrogen peroxide + ethylalcohol
=
acetaldehyde + acetic acid
ORG
(Newbee)
04-24-04 23:01
No 502720
Free for this week, Encyclopedia of Bioprocess Technology - Fermentation, Biocatalysis, and Bioseparation, Volumes 1-5: http://www.knovel.com/knovel2/Toc.jsp?Bo
(Fragile ego)
05-04-04 21:21
No 504822
You can make pyruvic acid with the dry distillation of tartaric acid which is OTC and cheap
Or you can search froogle for "calcium pyruvate"
http://www.democracynow.org/ - The Exception to the Rulers
(Stranger)
05-06-04 05:05
No 505189
[blue]But pyruvic acid may be useful in another way.
- pyruvic acid and diluted H2SO4 gives acetaldehyde.
[black]Well, if this is true, since the patent I was just reading claims that potassium bitartrate and H2SO4 mixed in equimolar ratios, yields 90% pyruvic acid when heated at 170C to 270C for one hour, can't one just use an excess of H2SO4, thereby turning this reaction into an acetaldehyde generator?
This sounds too easy and too OTC to be true?
Anyone going to try this before I get a chance?
Uncle Fester's Recipes suck
(Wonderful Personality)
05-06-04 11:08
No 505253
I cannot answer your question on the tartrate to acetaldehyde borolithium, sorry, but I must tell that FeSO4 + H2O2 + EtOH = acetaldehyde maybe theoretically nice but is practically a pain in the ass as the reaction tends to runaways.
Except one wants to go for real large amounts of product - kilos - I would suggest to skip the acetaldehyde, to concentrate on the production of benzaldehyde instead and to get a bigger bucket for the fermentation
Also I suggest to use diethylether for the extraction of the fermentation broth as the reductive amination/alkylation can be done directly onto the raw extract in ether - say: extract with ether, reduce the volume and do the Al/Hg in ether instead of alcohol.
(Stranger)
05-07-04 06:00
No 505504
It is not that hard to get you know. Where I am from, the grocery store will suffice. Granted, they are tiny bottles, but can't one just send a bunch of people out for the day?
IMITATION ALMOND EXTRACT
Also, bitter almond oil is not hard to get abroad. There are a thousand ways to smuggle it in, as I doubt there are any dogs that are trained to smell it. Shampoo Bottle?
That being said, it is my understanding that the presence of acetaldehyde is critical to the yield of L-PAC. Valuable benzaldehyde will be converted to benzyl alcohol in the absense of the acetaldehyde, and since acetaldehyde cannot be readily purchased, a simple and cost effective system of generating it is needed.
There are several patents detailing the use of acetylene, passed through a mercury or heavy metal catalyst at moderate temperatures.
Acetylene is available to all of us without raising any suspicion, but the mercury salts may be an issue.
Tartrates are OTC and would provide simple means of generating acetyldahyde without working with hard to obtain and dangerous ingredients, such as mercury salts.
Uncle Fester's Recipes suck
(Hive Addict)
05-07-04 11:48
No 505564
There are lots of ways to oxidise ethanol, for example pass ethanol/air over hot copper in a tube. Simple and cost-effective.
http://www.geocities.com/eric_vornoff/go
(Newbee)
05-09-04 07:22
No 505929
http://86.1911encyclopedia.org/P/PY/PYRU
When heated with hydrochloric acid to 10o C. it yields carbon dioxide and pyrotartaric acid C5H804, and when warmed with dilute sulphuric acid to 150 C it gives carbon dioxide and acetaldehyde.
This sounds waaaaaaaaaaaaaaaaaaayyyyyyyyyyyy easier than bulding some high temp, silver coated mesh catalyst burning tube, injecting ethanol and air streams in perfect proportions, and hopefully not having a big ass blow back or explosion. State of the art 19th century technology.
So, to simplify things:
Potassium Bitartrate + conc. H2SO4 in equimolar amounts, heated to 170C to 270C for 1 hour = 90% pyruvic acid
AND
Pyruvic Acid + dilute H2S04 heated to 150C = CO2 + acetaldehyde = essential ingredient for high yielding L-PAC
I would run the evolving gas through a cooling tube at 40C to condense any moisture, while allowing the acetaldehyde to mostly pass through. The gas can be bubbled into an aqueous solution, with conc. estimated by weight or volume. This solution can be later introduced into the bioreactor with the benzaldehyde to maximize the L-PAC yield.
Any objections?
Do Your Part To Help Stop Bovine Scatology
(Wonderful Personality)
05-09-04 10:13
No 505962
Any objections?
No.
Of course you will need some routine to get the very good yields told in literature (as always ), but it will work exactly like this. A powerful condensor for the acetaldehyde or bubbling it into ice-cold water is recommended.
ORG
(Newbee)
05-20-04 03:42
No 508310
A bioreactor seems to be the best way to maximize yields in a minimal amount of space. Constant aeration, agitation, and the slow introduction of the benzaldehyde and acetaldehyde are reported to provide the highest yield in the smallest amount of space.
Now if one was trying to purchase a bioreactor, there is only a handful of companies manufacturing these units, and they would likely ask what it was being used for, as these units are often customized. Can anyone think of a good, plausible industrial reason for wanting a bioreactor?
Do Your Part To Help Stop Bovine Scatology
(Fragile ego)
05-23-04 01:57
No 508869
You want to do some carefully controlled beer brewing experiments.
http://www.democracynow.org/ - The Exception to the Rulers
(Newbee)
05-23-04 03:20
No 508895
Thanks Foxy. I suppose that's too obvious. Mind you, I can't imagine any company getting too suspicious about a bioreactor.
Now here is a question for you foxy. as I remember a post you had put up eons ago regarding the strain candidus utilis. The majority of research papers I have come across using bioreactors use a strain of Sacharamyces Cerevesiae (I'm spelling this from memory so forgive me), a common yeast for white wine, for the conversion of benzaldehyde to L-PAC.
Is the candidus utilis (tarula yeast) superior in it's L-PAC/Benzyl Alcohol production ratio, and would it function much the same in a bioreactor as a substituted yeast strain?
Do Your Part To Help Stop Bovine Scatology
(Fragile ego)
05-23-04 05:08
No 508917
An interesting Brewing Chemistry article for you L-PACer's.
http://www.brewingtechniques.com/library
You can buy different yeast strains from a brewing yeast company that has high and low diacetyl strains. A high diacetyl strain might bump up yeilds nicely?
http://www.democracynow.org/ - The Exception to the Rulers
(Active Asperger Archivist)
05-23-04 18:15
No 509027
pyruvic acid synthesis from K bitartrate and Sulfuric acid
Post 384565 (cthulhujr: "Pyruvic Acid Synthesis and 24%", Stimulants)
Act quickly or not at all.
(Stranger)
05-28-04 22:42
No 510097
The pyruvic acid route to acetaldehyde sounds much more practical than from EtOH at high temperatures.
It's preparing an effective yeast that I have my doubts about.
The yeasts in industry are specially selected from colonies prepared. They are more resilient to B-aldehyde induced death. The hard-working ones are then extracted for the next experiment.
What about isolating yeast that survive certain treatments, and very small test-runs, then culturing those?
There's a method at Rhodium's using yeast from moldy apricots, and the yield from 264gm of B-Aldehyde was 24gm of L-PAC.
Not encouraging considering the level of skill in the art, that the experimenter probably has, and the equipment available.
Although no acetaldehyde was incorporated, or EtOH.
Here's that write-up: http://rhodium.ws/chemistry/ppa.l-pac.ra
How many liters of B-aldehyde is your average joe-bee going to go through before being comfortable with his procedure? For personal use I mean. (toluene will disappear.)
What B-aldehyde contaminents and by-products from it's preparation absolute no-no's for yeast, and which, if any, are acceptable?
How should air bee sterilized before introduction in to the bioreactor?
(Guardian)
05-28-04 23:41
No 510103
How should air bee sterilized before introduction in to the bioreactor?
Depending on the sensitivity of the reaction, usually using two in-line .2micron air sterilizing filters would work on the intake line. Or even placing an activated charcoal in- line filter upstream from the air sterilizing filters. If humidity would have to be introduced it can be done downstream from the .2 micron filters using water for injection as a source of humidity introduction. You can pick up the .2 micron air sterilizing filters for ~1.00 a piece USD for a pack of 100.
m_e
Unipolar Mania, It's good for life...
(Hive Bee)
05-30-04 16:55
No 510309
Adv Microb Physiol. 1999;41:1-45.
Factors affecting the production of L-phenylacetylcarbinol by yeast: a case study.
Oliver AL, Anderson BN, Roddick FA.
Department of Chemical and Metallurgical Engineering, RMIT University, Melbourne, Victoria, Australia.
L-Phenylacetylcarbinol (L-PAC) is the precursor for L-ephedrine and D-pseudoephedrine, alkaloids possessing alpha- and beta-adrenergic activity. The most commonly used method for production of L-PAC is a biological method whereby the enzyme pyruvate decarboxylase (PDC) decarboxylates pyruvate and then condenses the product with added benzaldehyde. The process may be undertaken by either whole cells or purified PDC. If whole cells are used, the biomass may be grown and allowed to synthesize endogenous pyruvate, or the cells may be used as a catalyst only, with both pyruvate and benzaldehyde being added. Several yeast species have been investigated with regard to L-PAC-producing potential; the most commonly used organisms are strains of Saccharomyces cerevisiae and Candida utilis. It was found that initial high production rates did not necessarily result in the highest final yields. Researchers then examined ways of improving the productivity of the process. The substrate, benzaldehyde, and the product, L-PAC, as well as the by-products, were found to be toxic to the biomass. Methods examined to reduce toxicity include modification of benzaldehyde dosing regimes, immobilization of biomass or purified enzymes, modification of benzaldehyde solubility and the use of two-phase reaction systems. Various means of modifying metabolism to enhance enzyme activity, relevant metabolic pathways and yield have been examined. Methods investigated include the use of respiratory quotient to influence pyruvate production and induce fermentative activity, reduced aeration to increase PDC activity, and carbohydrate feeding to modify glycolytic enzyme activity. The effect of temperature on L-PAC yield has been examined to identify conditions which provide the optimal balance between L-PAC and benzyl alcohol production, and L-PAC inactivation. However, relatively little work has been undertaken on the effect of medium composition on L-PAC yield.
Publication Types:
Review
Review, Academic
PMID: 10500843 [PubMed - indexed for MEDLINE]
.
05-31-04 03:41
(Rated as: UTFSE!)
(Chief Bee)
05-31-04 05:48
No 510457
Yes, everyone who UTFSE for it will be able to access it here: Post 505265 (Organikum: "Benzaldehyde -> L-PAC -> Ephedrine", Stimulants)
This article is also excellent: Post 507915 (Rhodium: "Benzaldehyde -> L-PAC -> PPA", Stimulants)
The Hive - Clandestine Chemists Without Borders
(Newbee)
05-31-04 09:17
No 510497
A charcoal filter alone will not sterilize the air in itself, as bacteria will breed quite easily on the charcoal surface. This is often a desired effect, such as in aquaculture, as the aerobic bacteria that colonizes on the charcoal will break down ammonia produced by fish waste into nitrite and ultimately into nitrate. The charcoal will only serve to absorb contaminates out of the air and will allow fungal spores or bacteria to pass easily through.
A HEPA filter or micron filter will catch most spores but not necessarily all of them, and not bacteria.
A UVC Sterilizer Lamp is a common option, available from hydroponic shops and water purification places. These bulbs sterilize the air or water that pass by them and will easily kill any bacteria or fungal spore. They do however, have the unfortunate effect of producing ozone, which may find it's way into the reactor and kill the substrate. If a charcoal filter is used after the UVC lamp, the ozone will be absorbed by the charcoal and %100 sterile air will pass through.
Another simpler option that I read from a brewing manual was having an air lock filled with bleach or alcohol. When suction is applied, air will be drawn through the liquid from one side of the air lock to the other, which will result in "sterile" air.
Just remember that there is a very distinct difference between contaminated air and sterile air, as contaminated air is often sterile and relatively clean air can still have microbes present. Charcoal only serves to decontaminate air and only certain contaminates at that.
Do Your Part To Help Stop Bovine Scatology
(Hive Addict)
06-02-04 20:21
No 510994
Well, the air temperature could be increased to the point that all or most bacteria would die. Then let cool for the reaction.
what about these ultrafilters (0.1µ) used for filtering pyrogenes and viruses?
What about buying a sterilizing filtering device/machine, used in industrial production, into which you install a membrane filter (cellulose nitrate).
These should not be expensive either.
(Stranger)
06-04-04 20:51
No 511482
How should air bee sterilized before introduction in to the bioreactor?
Use HEPA filters or bubble the air through EtOH. NOT denaturated alcohol of course.
(Wonderful Personality)
07-11-04 17:13
No 518701
It seems like acetaldehyde can be substituted by acetone in the biotransformation of benzaldehyde to phenylacetylcarbinol mediated by bakers yeast. This refers to the method using molasses, nutrient salts and thiamine (or whey, or brewers wort) and plain BAKERs YEAST (saccharomyces cerevesiae).
Acetaldehyde was applied as a hydrogen-acceptor in this reaction, its role was to scavenge hydrogen and to get converted to ethylalcohol. Every acetaldehyde converted to the alcohol says one benzaldehyde saved from being converted to benzylalcohol. (the main side product in this reaction).
It is long known that acetone is converted to isopropylalcohol by fermenting yeast (even mono and di-chloroacetone btw. - at least partially). The question if it will substitute acetaldehyde was not answered. Probably because it was never asked.
Now it was asked and answered all in one.
ORG
(Stoni's sexual toy)
07-11-04 17:23
No 518703
If acetone won't work (and your theory is correct), then maybe other, easier to handle aldehydes could be added?
BUSH/CHENEY 2004! After all, it ain't my country!
www.american-buddha.com/addict.war.1.htm
(Wonderful Personality)
07-12-04 18:53
No 518922
Acetaldehyde serving as hydrogen-acceptor is told in several articles and for example in the Patent DD51651 (some of the examples in the patent are proven to be working well btw.).
Acetone works, as with acetaldehyde the amount which can be added is limited due to its toxity to the yeast (benzaldehyde the same). Formaldehyde cannot be used for this reason - its plain too toxic. Acetone has the advantage not to kill the yeast (except in VERY high concentrations but only to suppress fermentation. After some time the yeast will recover as the processes which transform the acetone to the alcohol (which is also toxic but much less) and the benzaldehyde to the l-PAC are independent from fermentation. The fermentation replentishes enzymes which are used up though.
A lower toxity substitute would be preferred of course. Any suggestions?
(Hive Bee)
07-12-04 23:20
No 518974
Pyruvic acid maybe?
I'm not in the mood to go trough the whole thread, but aren't pyruvates used in some patents? Pyruvic acid is surely less toxic than acetone and acetaldehyde, besides being quite OTC and yielding only the edible lactic acid as a side product. Another less toxic ketone should be 2-butanone, but it is less soluble in water.
“The real drug-problem is that we need more and better drugs.” – J. Ott
(Newbee)
07-12-04 23:37
No 518980
Yes, one of the patents I liked used pyruvic acid and the yeast saccharomycae cervisiae (sp?). Pyruvate is very OTC, too.
--aia
(Wonderful Personality)
07-13-04 03:47
No 519028
Look:
As you can see, the pyruvic acid is the reactand which produces l-PAC with benzaldehyde. If the pyruvic acid is added directly or if it is produced by the yeast from molasses or glucose doesnt make so much difference. If pyruvic acid or a salt of it is added directly non-fermenting conditions are used, if molasses or another source of fermentable sugars is used activly fermenting conditions have to be used. But this nothing to do at all with the question for a hydrogen acceptor, which plays solely the role to act as additional prey for the also yeast mediated aldehyde to alcohol reaction.
This for acetaldehyde is usually added which ends up as ethylalcohol.
Also ethylalcohol can be added to the biosynthesis to suppress ALL alcohol producing processes mediated by the yeast as far as possible. Yeast reduces them when a certain level of alcohol is reached in an attempt not to commit suicide.
I hope I could clear this up.
ORG