08-30-01 23:36
No 208763
The concept of utilizing this beautiful fungus has been burning in the backs of my mind for a while. There have been quite a lot of threads regarding this in a lot of forums.
The biosynthetic pathway of the fungus is as follows:
Tryptophan --+tryptamine --+ nmt --+ dmt --+psillocin --+psilocybin.
A bit more goes on here, though.
OK folks, I'm going to name some enzymes. I'm probaly not going to do this right, and I'm not 100% sure the coonvention, or if any of these have been named. PLEASE CORRECT ME O WISE AND KNOWNING ONES (not sarcastic)
Each of these steps is contrlled by an enzyme (as well as co-enzymes), and every enzyme is controlled by a single gene. One gene, one enzyme.
So tryptophan is animated by the enzyme tryptophan animase, then it is methylated by the action of tryptamine methylase, twice, rather than by tryptophan dimethylase, as is confirmed by the presence of baocystin. The enzyme tryptamine 4-hydroxylase hydoxylates the dmt creating psilocin, and the nmt creating the precursor to baeocystin, 4ho nmt. The psilocin and 4ho nmt are then the substrate for 4 ho (tryptamine) phospholase, creating, respectively, psillocybin and baeocystin.
It could be that 4 ho tryptaminase requires mono methylation of the nitrogen. This could be determined by the presence of 4-hydroxytryptamine, if present in the fungus, it would prove that any non-methylated tryptamine could be hydroxylated in the 4pos through the use of this biosynthetic enzyme. This would provide an interesting avenue for the synthetic chemist, both as a direct agent of synthesis and as a launching pad for new compunds.
It would be just as interesting to find 4 phosphorloxy tryptamine, which should be present if the hydroxy is.
Unlike most organism that break down their food-molecules, fungi pull them in whole. This unique property allows the placement of substances within the growing medium that are drawn into the organism and fixed in the metabolism of the mushroom, with the creation of new substances. Their isolation is the most difficult stumbling point right now, and would seem to require specialized equipment. I believe that the isolation of the 4ho's would be possible, given their polar-freebase, in contrast to most alkaloids, but I degress. By placing, say, 2-flurotryptamine (just a bs example), we would end up with 2-fl,4-ho,t. The requirement seems to be that in order for the fungus to fuse a 4ho, there can't be anything in the 4 or 5 position, and that it be a tryptamine. Beyond that, the sky's the limit.
DPT__>4 ho dpt
Dipt__>4 ho dipt
Amt__>4 ho amt ETC
But really you get a lot of shit, because the normal biosynthesis is still happening.
If a culture could be isolated (through mutagenesis) that did not produce psillocybin, it would be easy to determine which genes were lacking. I do not know if this is actually possible, anyone know what metabolic processes are effected by psillocybin?
If the culture produce psillocybin given either tryptophan, tryptamine, nmt, dmt, or psilocin, it would be easy to determine which relitive gene is affected, and thus the enzymes related.
The modification of the genome could prove to be the most powerful tool of synthetic chemestry of the future. What if one nucleotide alteration produced the enzyme isoprpylase? You'd have mushrooms making 4 ho dipt. Perhaps isolation of the enzyme typtamine 5 methoxylase from Phalaris would allow a mutagenic cross. Mushrooms containing foxy methoxy, anyone? The possibilities are literally endless.
Looking at the slew of alkaloids in peyote makes me horny. Imagine the enzymes available there! Or in other legal plants and mushrooms! Perhaps there's a fungus out there that is churning out 4 ho nndmtryptophan, which would produce psillocybin given a trypamine substrate.
Well, enough ramblings 4-now.
Today is opposites day. Everything I say, I mean the opposite.
(Newbee)
08-31-01 22:20
No 209098
So tryptophan is animated by the enzyme tryptophan animase
Huh? I thought tryptophan was decarboxylated to tryptamine by tryptophan decarboxylase. There's no way getting tryptamine from tryptophan by aminating it.
understanding is everything
(Ubiquitous Precursor Medal Winner)
09-01-01 05:18
No 209213
Understanding is everything. Tryptamine methylase, twice, sounds like a most useful enzyme, more than half as useful as tryptamine dimethylase. (Real stuff, I take it.) Perceptive of you to spot (and emphasize) the fungus will process substituted tryptamines.
turning science fact into <<science fiction>>
(Stranger)
09-06-01 01:07
No 210483
Your right, of course, tryptophan decarboxylase.
Thanks, and why didn't the rest of you catch me?
Now, what to do with 700 quart mason jars . . .
Today is opposites day. Everything I say, I mean the opposite.
(Hive Bee)
09-11-01 05:19
No 211948
I was interested in this while at my previous employer. A small life crisis measuring 6.0 on the 'oh shit' scale, and I postponed the idea. But here is what I found: See my original Post 173127 (KrZ: "Re: PSILOCIN Enzyme Sequences", Tryptamine Chemistry)
KrZ-
I wasn't able to get any of the articles, but I'm not sure of their worth anyway. (since they all pre-date molecular bio/DNA techniques)
Ironically, the textbook Biochemistry by Mathews/vanHolde Chapter 21 page 740, review question 4,actually asks for a biosynthesis of psilocybin from tryptophan. Too bad there's no answers in the book.....
Regardless, you stated:
Tryptophan -1-> Tryptamine -2-> N-Methyl-tryptamine -3-> N,N-dimethyltryptamine -4-> psilocin -5->psilocybin
The enzymes for the first 3 steps are known, and the protein+DNA sequences are known too.
1-Tryptophan->Tryptamine
Enzyme=aromatic amino acid decarboxylase
AKA: AAAD, AKA: dopadecarboxylase
Genbank Accession# M88700 (protein sequence=AAA20894)
Reference:Human dopa decarboxylase: localization to human chromosome 7p11 and characterization of hepatic cDNAs. Genomics 13 (2), 469-471 (1992)
2-Tryptamine -> N-Methyl-tryptamine
Enzyme=Indolethylamine-N-Methyltransfera
AKA INMT
Genebank Accession# NM_006774
Reference:Human indolethylamine N-methyltransferase: cDNA cloning and expression, gene cloning,and chromosomal localization. Genomics 61 (3), 285-297 (1999)
3-N-Methyl-tryptamine -> N,N-DiMethyl-tryptamine
Same as #2
4-N,N-dimethyltryptamine -> psilocin
There most likely isn't a human gene for this. (otherwise we'd all be alot happier people)
I find no references in medline for: indole-hydroxylase, trptamine-hydoxylase, or indolamine-hydroxylase, leading me to believe there is no human (or other species) homolog. Dead end? My first thoughts are maybe the psilocin step is impossible at this time, or at least until someone clones the gene responsible. So where from here? Perhaps the 5-hydroxyderivative would be more attainable.
See 5-OH-N,N-DMT http://www.erowid.org/library/books_onli
Shulgin is less than enthusiastic about this compound.
This could be accomplished with tryptophan hydroxylase. Worth the effort?? I'm skeptical.
5-psilocin -> psilocybin
Haven't looked yet.
That's all my ramblings. Any feedback would be much appreciated.
Cheers.
-quicksilver-
(Stranger)
09-17-01 23:20
No 214474
Well, I'm sure you want feedback from anyone but me (as I am lacking true knowledge), but what about 4-acetoxylating dmt rather than hydroxylating it? The acetoxy group is converted into the hydroxy (supposedly), and seems a bit more commen, although the 4 pos isn't.
I've also wondered why no one's ever extended the chain, you know, 4-methoxy,n,n-dmt etc. Here in the hive, I'm the second dumbest guy around (you know that one guy who keeps asking about rp under all those names), so I figure if I flame myself than everyone else can get back to work.
I looked around medline for indole-acetoxylase, tryptamine-acetoxylase, enamine-acteoxylase (that's right, right? enamine=indole?) and indolamine-acetoxylase, but I could be looking right at it and not see it.
Today is opposites day. Everything I say, I mean the opposite.
(Hive Bee)
09-19-01 05:39
No 214987
>what about 4-acetoxylating dmt rather than hydroxylating it?
>The acetoxy group is converted into the hydroxy (supposedly), and seems a bit more commen, although the 4 >pos isn't.
Converted how and where?
>I've also wondered why no one's ever extended the chain, you know, 4-methoxy,n,n-dmt etc.
Innovation is done by the few. Bee a pioneer! Synth something novel! I would assume it has some activity. Maybe something between psilocin & psilocybin.
BTW, Shulgin did make N,N-Dimethyl-4,5-methylenedioxy-T.
(Hallucinogenius)
09-19-01 06:06
No 214992
On the distribution of recombinant organisms; I had a discussion about psilocin biosynthesis in the tryptamine forum some time back. E. Coli. has been discussed as a potential expression system, which is all well and good, but why try switching from eukaryote to prokaryote and risk any possible disulfide bridges and glycosylations? An edible plant expression system might work well, but is more complicated than yeast expression... But how to get the genes out of P. Cubensis? (which appears to be so good at the pathway, since it can produce 3% by dry weight of these products) Well. A quick GenBank search for tryptophan decarboxylase turned up some hits. 6 to be exact. While this is pretty minimal, a BLAST search turned up some interesting stuff. It seems that among the tryp./DOPA/tyrosine decarboxylases, the large large majority (A dozen or so across numerous species) show this highly conserved sequence; gacgcagcttacgc. Generally we would prefer an 18 base probe, but 14 will still work pretty well, with an average occurence of 4^14 it should be relatively specific. Some additional BLAST work on INMTs showed a conserved sequence among several divergent species. This one is even longer, located near the middle of most of these cDNAs; tcctgcgtggccctggagaa. The genome of Aspergeillus Nidulans is around 28 mb, I have no earthly idea how big P. Cubensis' is, but lets assume it's 150 mb just to be safe. From N=1-(1-f)^P, we would need 15000 cosmids (45kb) to cover at 0.99 P. Then probe, isolate those transformants with our gene of interest, and check their cDNA products. Anyway, the DMT producing enzymes in the pathway should be relatively easy to isolate. Affinity binding, protein sequencing, and tRNA preference/low redundancy (or 0) regions could further elucidate the pathway enzymes. I have a friend in mycology who does nothing but make DNA libraries all day, and she seemed to find the idea sound, although quite time consuming for a renegade anarchist Humans deserve the right to ingest and experiment with whatever compounds they like, and this really takes it to the non-scientists. This general schema could be applied to numerous other natural psychadelic products, tyrosine to mescaline, DMT, psilocin, psilocybin, amphetamine/methylamphetamine (bleah, not worth the trouble), etc.. What others can you think of? I know that these products can be cultured already, from their natural hosts, and subsequent extraction, but this opens up alot of possibilities, not all of which are expressed in sufficient quantities to be practical/easily grown. I pulled out some of the best parts for you quicksilver, neat stuff; Psilocybin biosynthesis 4-Hydroxylated or 4-methoxylated indoles are very rare in nature. The only known examples beside the psilocybin-type alkaloids are the 4-hydroxylation of indole acetic acid by Aspergillus niger strains (54), methoxylated b-carbolines from Banisteriopsis argentea and Picrasma javanica, the reserpine analog venenatine from Alstonia venenata, the yohimbine analog mitragynine from Mitragyna speciosa (79, 82 p. 703), and the aminopyrimidyl-indolic meridianines from the tunicate Aplidium meridianum (25). Psilocybin has a simple structure in contrast to the former alkaloids. It is formally derived from tryptophan in five distinct biosynthetic reactions, i.e. decarboxylation, indole-hydroxylation, two N-methylations, and O-phosphorylation (Table 1). Feeding experiments with putative intermediates, analogs of them, or radioactive precursors supported the view that tryptophan decarboxylation is the first biosynthetic step and that O-phosphorylation is the final step. The sequence of the remaining intermediate reaction steps is still unclear. Some authors even suppose a biosynthetic grid with multiple routes to psilocybin (Figure 1) (13, 5, 78). Abbreviation Full name Decarboxylase Tryptophan Decarboxylase Hydroxylase Tryptamine 4-Monooxygenase, Tryptamine : Oxygen Oxidoreductase, 4-hydroxylating Methylase Tryptamine-w-amino-methyltransferase Phosphorylase Psilocin-O-phosphotransferase Table 1. The psilocybin synthesizing enzymes. The full name follows standard enzyme nomenclature while the abbreviated name is used throughout this text. The enzymes may actually consist of several related enzyme with slightly different substrate specifities. Expression cloning procedure For a definitive evaluation of psilocybin biosynthesis, the participating enzymes need to be isolated and tested for their substrate specifities and activities. The feasible way to do this was by genetically cloning and heterologous expression. The cloning methodology chosen is an alternative to PCR based procedures and has been devoloped and used succesfully to clone a broad range of fungal enzymes (21, 20, 15, 14, 53). It consists of ligating a cDNA library into an expression vector (pYes2), transforming a host organism (S. cerevisiae), and a screening the resulting colonies for enzyme activity (Figure 2). The success of such expression cloning procedures depends on reliable and sensitive enzyme assays for the colony screening step. Indeed it was possible to find such assays for all enzymes of the psilocybin biosynthesis pathway (Table 2). A second important requirement is the use of biomaterial containing high amounts of the respective enzymes mRNA. Usually this is a growing tissue containing the metabolites of interest. In this study still developing Psilocybe tampanensis sclerotia were used for mRNA extraction. They grew rapidly and reliably on a special medium and contained high amounts of psilocybin and psilocin. Total RNA isolation from enzyme producing biomaterial \/ mRNA preparation \/ cDNA synthesis \/ Ligation into E. coli / S. cerevisiae shuttle expression vector (pYes2) \/ Amplification in E. coli \/ Colony pooling and plasmid preparation (20 pools of 5,000 colonies each) \/ Yeast transformations \/\/\/\/ Enzyme activity screenings of colonies by color reactions after inducing expression (galactose) \/ Rescreening positive clones \/ Cross-transformation of E. coli, insert sequencing and analysis Figure 2. Expression cloning flow scheme. As used by Dalbøge et al. to clone a broad range of fungal enzymes. Enzyme Screening procedure Decarboxylase 5-fluoro-tryptophan resistance Hydroxylase feeding tryptamines, Keller's reaction Methylase feeding tryptamines, derivatization and removal of substrate, radioactive detection Phosphorylase feeding psilocin, derivatization of substrate, Keller's reaction after phosphatase treatment Table 2. Enzyme activity screening procedures. All tests can be performed on colonies to allow a parallel screening procedures. The Keller's reaction is a very sensitive and specific color reaction for hydroxylated indoles. Mushroom media and culturing Media ingredients and sources: dried unrefined sugar-cane extract (Rapadura, organic food or third world stores), sugar-beet syrup (75% dry matter, food stores), mixed pollen (organic or health food stores), dry yeast extract (Oxoid, England), peptone (Difco Laboratories), agar (Merck, Germany), commercial malt extract / yeast solutions "Salvator" (containing 18.3% stammwuerze) and "Hefe Weissbier dunkel" (dark unfiltered wheat beer, containing 12.4% stammwuerze, both from Paulaner, Munich, Germany). The mycelia were cultured on Parafilm sealed MEY plates (6% malt extract syrup, 0.6% yeast extract, 1.5% agar) at 28°C ± 2°C in an incubator (6). For propagation 1 cm x 1 cm blocks were cut out and transferred to the middle of a new agar plate under sterile conditions. Plant hormones were added as ethanolic solutions. Media containing KH2PO4, pollen, acetic acid, citric acid, and ascorbic acid medium were autoclaved after adjusting the pH. Media from beer (pH around 5.5) were autoclaved for 40 min. Mushroom extraction For a simple but efficient extraction of psilocin as well as psilocybin the following method was applied (98, 58, 45). 7.5 mg lyophilized mushroom material was homogenized in a 1 ml glass mortar with 250 µl methanolic extraction solvent (75% MeOH, 0.1% ascorbic acid). This suspension and 2 x 125 µl methanolic rinsings were pooled in a microcentrifuge tube. After agitation for 10 min at RT the tube was centrifuged for 2 min at 14,000 rpm at RT (5415C centrifuge), the supernatant was transferred to a fresh tube, and the pellet was resuspended in 250 µl ethanolic extraction solvent (75% EtOH, 0.1% ascorbic acid). After agitation for 10 min at RT and centrifugation as above the supernatants were pooled and stored at -20°C in an airtight microcentrifuge tube. Reagents and procedures from the cDNA Synthesis System Kit (Gibco BRL) were used in combination with a vector primed cDNA synthesis literature method (85). This approach was not succesful. Here's a cool transformant screening procedure; 5-Fluoro-tryptophan decarboxylase screening For tryptophan decarboxylase screening transformed FY 73 cells were plated directly onto SC-gal containing 0.5 - 1.5 mM 5-fluoro-tryptophan and were incubated up to 6 days at 30°C. No-Go for submerged culture; The tested Psilocybe strains showed variable preferences for the tested media. In general, the defined Leatham’s media, sugar-cane medium, and beer-based media were poor substrates compared to malt extract based media. The submerged culture produced the highest amounts of biomass, followed by the surface cultures on soft agar (0.2%). But under both conditions the mycelia did not produce measurable amounts of alkaloids. The same was true for Ps. tampanensis submerged cultures in 6% malt extract with no supplement, 0.3% yeast extract, 2% pollen, or both added (data not shown). Ps. cubensis, Ps. tampanensis, Ps. azurescens, and Ps. cyanescens were grown in different media. After 14 days the mycelium was harvested, weighed, extracted, standardized, and analyzed for psilocin and psilocybin content. Ps. cubensis produced psilocin, but no psilocybin under the conditions analyzed. In contrast Ps. tampanensis and Ps. azurescens produced both alkaloids. Ps. azurescens and Ps. cyanescens were growing very slowly on all media tested. Psilocin and psilocybin Rf values Psilocin and psilocybin references (in slightly acidic solution, isolated from Ps. cyanescens fruiting bodies) were TL-chromatographed using various solvent systems. The plates were stained with Van Urk’s reagent (DMCA modification) and Rf values were observed (Table 8). Solvent system Psilocin Rf Psilocybin Rf H2O: MeOH: AcOH (50 + 50 + 1) 0.92 0.88 MeOH 0.54 0.11 Acetone 0.30 0.06 MeOH: AcOH (1 + 1) 0.33 0.13 H2O 0.88 0.47 H2O: AcOH (1 + 1) 0.88 0.76 nPrOH: H2O: AcOH (10 + 3 + 3) 0.71 0.39 CH2Cl2 0.00 0.00 Best Host; Yeast as host organism An important factor for a succesful expression cloning procedure is the choice of an adequate host organism. Yeasts, especially S. cerevisiae, are easy to culture and grow rapidly. As well efficient transformation techniques in common with bacteria have been developed. Otherwise they have many compartments and posttranslational processing and transport systems in common with complex multicellular eukaryotes, especially with other fungi like mushrooms. For the crosstransformation from positive clones into E. coli simple methods exist (90, 84). REFERENCES 1. Brassinosteroids. G. Adam, V. Marquardt. 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Miozzari, P. Niederberger, R. Hütter. Anal. Biochem. 90: 220 (1978) 71. Interrelationship of phosphate nutrition, nitrogen metabolism, and accumulation of key secondary metabolites in saprophytic cultures of Psilocybe cubensis, Psilocybe cyanescens, and Paneolus campanulatus. J. M. Neal, R. G. Benedict, L. R. Brady. J. Pharm. Sci. 57: 1661 (1968) 72. Pharmacotheon. J. Ott. Natural Products Co., Kennewick, WA (1993) 73. Transformation of E. coli using homopolymer-linked plasmid chimeras. S. L. Peacock, C. M. McIver, J. J. Monahan. Biochim. Biophys. Acta 655: 243 (1981) 74. Determination of psilocybin in Psilocybe semilanceata by capillary zone electrophoresis. S. Pedersen-Bjergaard, E. Sannes, K. E. Rasmussen, F. Tønnesen. J. Chrom. 694: 375 (1997) 75. Psilocybian mycetismus with special reference to Paneolus. S. H. Pollock. J. Psyched. Drugs 8: 43 (1976) 76. GLC-mass spectral analysis of psilocin and psilocybin. D. B. Repke, D. T. Leslie, D. M. Mandell, N. G. Kish. J. Pharm. Sci. 66: 743 (1977) 77. Baeocystin in Psilocybe semilanceata. D. B. Repke. J. Pharm. Sci. 66: 113 (1977) 78. Baeocystin in Psilocybe, Conocybe and Paneolus. D. B. Repke. Lloydia 40: 566 (1977) 79. Psilocin analogs. III. Synthesis of 5-methoxy- and 5-hydroxy-1,2,3,4-tetrahydro-9H-pyrido[3
(Hive Bee)
09-20-01 06:42
No 215397
Now that's a LT-persona sized block-o-text you got there! Let me take a bite out of it/ask some questions.
Why use e coli?
I agree. Why not yeast. Or rice!
This general schema could be applied to numerous other natural psychadelic products, tyrosine to mescaline, DMT, psilocin, psilocybin, amphetamine/methylamphetamine (bleah, not worth the trouble), etc.. What others can you think of?
Brewers yeast that makes safrole, ephedrine, etc. Post 27698 (user1634: "Not insects! Trees!", Novel Discourse)
Psilocybin biosynthesis 4-Hydroxylated or 4-methoxylated indoles are very rare in nature. The only known examples beside the psilocybin-type alkaloids are the 4-hydroxylation of indole acetic acid by Aspergillus niger strains (54), methoxylated b-carbolines from Banisteriopsis argentea and Picrasma javanica, the reserpine analog venenatine from Alstonia venenata, the yohimbine analog mitragynine from Mitragyna speciosa (79, 82 p. 703). ref#54 -no enzyme mentioned. No mention in #82 (TIHKAL) or #79 I assume?
Seeing as how these enzymes are conserved across species.
It may be possible to skip all the work of getting the mushroom enzyme and instead just using the human or mouse versions.? Maybe. I dunno.
(Hallucinogenius)
09-20-01 07:14
No 215409
I would bank on the fungal enzymes being far more active, FAR FAR more. Not to mention easier to isolate, less likely to have unwieldy post-translational modifications, and more likely to exhibit matching tRNA preference to any expression system one might choose. If the conservation we see is an accurate depiction of the Psilocin genes, then it would be easier too, just because the human genes have been sequenced doesn't mean they're easy to get at!
(Newbee)
09-20-01 16:11
No 215573
This is what (mostly) inspirid the 4acetoxy/4mydroxy conversion comment
---
Recently, I had the opportunity to experiment with the more stable 4- acetoxy-DIPT derivative. It is theorized that once a 4-acetoxy compound enters the body it gets chopped off by esterase enzymes, leaving behind the active 4-hydroxy group to work its magic on the brain. It s very similar to how 4-Phosphoryloxy-DMT (psilocybin) is metabolized into 4-HO- DMT (psilocin). For all intensive purposes the effects of isolated psilocybin and psilocin have been found to be subjectively the same. This can also be said for 4-acetoxy-DMT, 4-acetoxy-DET, and their 4-HO counterparts. In my first comparison experiment, 4-HO-DIPT and 4-acetoxy- DIPT were found to be essentially the same experience, and others have reported the same conclusion.
It's from toad 4 aces primer. Also there is a research chem supplier whos name sounds like "a shoe" that seems to be running with the idea. They sell, 4-acetoxy,dipt; 4 ace., det, etc.
About bee-ing a pioneer, someday. I still have a bit to learn, I'm not comfortable bluffing in this circumstance, and I'm plodding through Chem 2 right now. Any ideas on the subject would be welcome
Add 5meodmt to the list of possibles. Not that great of an adddition, but its something.
Today is opposites day. Everything I say, I mean the opposite.
(Hive Addict)
09-26-01 06:43
No 217266
"because the normal biosynthesis is still happening.
If a culture could be isolated (through mutagenesis) that did not produce psillocybin, it would be easy to determine which genes were lacking."
This is not that easy my friend. So you dose your spores with enough EMS to give your grandma a bp mutation. O.K. , mutagenesis gives a pretty damn low rate of mutation, so thousands and thousands, and, did I mention thousands, of separate cultures would get growing. Then you screen them. Then you search through a psilocybe or sumsuch genome, which is probably not mapped because no one wants to fund it. After a years work of constructing polymorphic markers , you have narrowed your region, them bamm! What the hell, that is a 60s ribosomal transcription gene, back to the old drawing board. Easy no, possible, very. All you need is a lab and a few hundred thousand dollars.
4:20
(Newbee)
09-27-01 20:35
No 217819
That's all? I thought that it would bee hard . . .
Today is opposites day. Everything I say, I mean the opposite.
(Hive Bee)
10-07-01 04:23
No 221080
Poster: KrZ
Subject: Re: Entheogenomic Research
I would bank on the fungal enzymes being far more active, FAR FAR more. Not to mention easier to isolate, [snip]
I see this differently. I think since DMT occurs naturally in humans, that our enzymes are perfectly capable. The 4-hydroxylation to psilocin might be another story though. As far as being easier to isolate, I imagine there are more researchers working with human DNA libraries than those working with P Cubenis libraries.
(Hallucinogenius)
10-07-01 10:38
No 221193
No, you are absolutely wrong. NO NO NO NO NO.
(Hive Bee)
10-08-01 05:29
No 221498
Poster: KrZ
Subject: Re: Entheogenomic Research
No, you are absolutely wrong. NO NO NO NO NO.
I think i just got schooled by a hallucinogenius.
Don't you want to back that theory up with a 1000-word block of unpunctuated text?
(Hallucinogenius)
10-08-01 08:36
No 221567
I'll just say this, post-translational modification, tRNA preference, ease of library preparation, genome size, and NORMAL LEVELS IN HOST.
(Stranger)
01-14-02 01:09
No 256055
http://meltingpot.fortunecity.com/gregor
"Attempted Molecular Cloning of Enzymes from the Psilocybin Biosynthesis Pathway in Psilocybe tampanensis"
Almost 60 pages of extra tasty goodness. Outlines the enzyme pathways as well as attempts to genetically engineer yeast and E. Coli with these pathways. Lots of information on feeding the magic fungus precursors but no mentions of DIPT.
So has anyone revealed empirical data on DIPT -> 4-HO-DIPT via biosynthesis?
-Zen
(Hive Bee)
01-21-02 15:57
No 258990
Your link is broken, Zen.
(Moderator)
01-21-02 17:38
No 259015
I removed the linebreaks from the link, it should work now.
(Ubiquitous Precursor Medal Winner)
01-21-02 18:07
No 259026
as in http://meltingpot.fortunecity.com/gregor
turning science fact into <<science fiction>>