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Coincidentiaoppositorum
deep psychedelic

Registered: 10/27/14
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Sorry that post got all jumbled, technical issues with my device...
-E. Borodin
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Coincidentiaoppositorum
deep psychedelic

Registered: 10/27/14
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Abstract Mycelial cultures of PSILOCYBE CUBENSIS, with the ability to form psilocybin and psilocin DE-NOVO, also hydroxylated and methylated fed tryptamine to give psilocin in up to 3.3% dry mass of the obtained fruit bodies. By using HPLC and TLC, it was found that these mushrooms contain only a small amount of psilocybin (0.01-0.2% dry mass). The values of psilocin are the highest described in any mushrooms. - http://www.ncbi.nlm.nih.gov/pubmed/17262409
This says the tryptamine inoculate substrate produced cataphores with low psilocybin, but the highest psilocin rate observed
(Psilocybin is a pro-drug, the 4-phosphorloxy group is hydroxylated and it becomes psilocin in vivo shortly after ingestion....so low psilocybin is not bad, however psilocybin is more stable as far as degradation...regardless very interesting stuff...
I always figured a substrate high in tryptophan would be sufficient....thanks for getting into this topic, I have actually learned a good deal.
-E. Borodin
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Coincidentiaoppositorum
deep psychedelic

Registered: 10/27/14
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Last seen: 8 years, 4 months
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It drives me crazy to talk about biosynthetic pathways when I cant see whats actually going on, so for all you other visual learners out there, I hope this helps.
-E. Borodin
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Toadstool5
A Registered Mycophile



Registered: 01/22/15
Posts: 1,359
Loc: The Golden State
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Re: Tryptamines in substrate with DPT? [Re: micro]
#22238504 - 09/14/15 08:30 PM (8 years, 4 months ago) |
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Quote:
solubility matters as there are plenty of insolubles the mycelium can digest
Solubility is a bad word for it. I'm referring to how well the precursors are absorbed by the mycelium. Perhaps permeability is a better word?
If only there was more literature on this subject 
Great diagram on the pathway!
-------------------- If you do not know where the mushroom products you are consuming are grown, think twice before eating them. - Paul Stamets AMU Teks Stro's Write Ups
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Coincidentiaoppositorum
deep psychedelic

Registered: 10/27/14
Posts: 1,965
Last seen: 8 years, 4 months
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Re: Tryptamines in substrate with DPT? [Re: Toadstool5]
#22238673 - 09/14/15 09:10 PM (8 years, 4 months ago) |
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Here's some links that may be helpful, ill be more willing to discuss my veiws on the matter in the morning when I have got some rest.
http://www.lycaeum.org/mv/mu/cubensis_parameters.html this link is great general information, really good stuff, highly recommended.
http://www.researchgate.net/publication/6542579_Biotransformation_of_Tryptamine_in_Fruiting_Mycelia_of_Psilocybe_cubensis Biotransformation link.
http://onlinelibrary.wiley.com/doi/10.1002/jobm.3620290608/abstract;jsessionid=C4069DF287E79A79E4B17E9DDB0708E9.f02t02 Biotransformation of tryptamine derivatives in mycelial cultures (psilocybe cubensis) by gartz
onlinelibrary.wiley.com/doi/10.1002/jobm.3620290608/pdf
http://chemistry.mdma.ch/hiveboard/tryptamine/000127125.html Stuff from the hive on the topic
http://www.stainblue.com/ah.html Here's some hoffman information
https://www.erowid.org/archive/rhodium/pdf/ psilocybin.biosynthesis-2.pdf And an erowid PDF on psilocybin biosytnthesis.
https://www.erowid.org/culture/characters/gartz_jochen/gartz_jochen.shtml here's erowids gartz page, there's some great links in it
https://www.thevespiary.org/talk/index.php?topic=12665.10;wap2 (Everything below came from the link.above) 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 x1 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 x125 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
-E. Borodin
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Coincidentiaoppositorum
deep psychedelic

Registered: 10/27/14
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I'm not sure how specific any of that information was, but it kept me interested.
-E. Borodin
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micro
bunbun has a gungun



Registered: 05/09/03
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Quote:
Coincidentiaoppositorum said: Mycelial cultures of Psilocybe cubensis capable of forming psilocybin and psilocin de novo display a high capacity for hydroxylation of tryptamine derivatives at the 4-position.
High capacity =/= everything. I guess it was just taken out of context.
They have isolated those enzymes? I wonder if they have been sequenced or just ID'ed on western blot or something and then named?
Only reason I ask is it would be funny to TA-clone them into S. cerevisiae, assuming one had standard molbio resources and the ability to purchase the primers and a kit (if they aren't stocked). It would take me half a day (mostly waiting) to run the PCR and prolly get the rest done that day, culture overnight, transfer to TB/LB media, etc. I don't work in molbio anymore, though.
EDIT: Oh wait, looks like they did that.
I want some culture of that 
Interesting info; I'll have to check it out in depth when I have a bit.
-------------------- Any research paper or book for free (Avatar is Maxxy, a character by Mizzyam, RIP)
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Coincidentiaoppositorum
deep psychedelic

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Re: Tryptamines in substrate with DPT? [Re: micro]
#22240628 - 09/15/15 10:19 AM (8 years, 4 months ago) |
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The biosynthesis of psilocybin in psychedelic mushrooms is a multi-step process, and the precise mechanism is debated by many authors...-countyourculture
Experiments with radiolabled precursors have shown that this is likely the primary path to psilocybin,however, labelled 4-hydroxytryptamine was also shown to be incorporated into the produced psilocybinindicating the possibility of an additional biosynthetic pathway. Other alkaloids present in psilocybin mushrooms such as baeocystin or norbaeocystin are not explained by this single pathway as well. An elegant alternative has been proposed. What if instead of a single path and a set order of modifying reactions, there were multiple paths to psilocybin – with branching edges that led to baeocystin and norbaeocystin? The enzymes would compete and feed back among each other in a biosynthetic grid that preferred to produce psilocybin and psilocin but also produced small amounts of baeocystin andnorbaeocystin as typically seen in nature. -countyourculture
They are refering to this pathway: -tryptophan--tryptamine--DMT-psilocin-psilocybin
I know dimethyltryptamine better than psilocin...so ill start with what I know about its enzamatic pathway...
Which starts from tryptophan which is decarboxylated (amino acid decarboxylase, AADC)
Next we have tryptamine which is met with indole-amine methyl transferase (INMT) and exposed to s-adenosyl-methionine. (SAM) which becomes s-adenosyl-l-homocystine (SAH) as it adds the methyl grouping.
First time we get NMT, second round DMT...
Then its enzamatic hydroxylation then 0-phosphorlation to psilocybin
As far as isolation of the enzymes, yes, it has happened, and work is being done. But molecular biology is not my strong point, I'm around 3rd year organic chemistry in my knowledge and skill level here, which means anything in TIHKAL or PIHKAL is a breeze...it gets a bit more complicated with Albert hoffman as far as techniques and equipment goes, but its still nothing a second or third year organic chemistry student couldn't figure out.
And that's where I spend most my time, in the study of Sasha shulgin, Albert hoffman, David E. Nichols, Daniel Trachsel and even some of the ethnobotanists like schultes or wasson, and I love to understand how these things are acting in our body or formed in nature, but in all honesty its the organic chemistry work of people like shulgin, heffter, Nichols, hoffman, etc..that fascinates me, and that relates to my educational goals.
It looks like you have your work cut out for you though, molecular biology is a very demanding field of study, I have not looked into it much, well any more than I would have to, and since I'm a person who enjoys recipe oriented activities and math, org. Chem just seemed to really fit, where biology, physiology, and things of this nature, seemed like things I had to learn because of their relation to something I cared about, and I see them more as a pain in the ass than fun, but that's why science is divided into specialized fields.
So sorry, I wish I could help, but your getting a bit out of my range of study, and though it does involve tryptamine chemistry, there's probably around 100 different active tryptamines just covered by TIHKAL alone, so ive only gotten really in depth on a few of them, and I do know a good deal about psilocybin, but its just one of many, plus I have only been in study of these compounds for 2.5-3 years, I still have a good deal to learn.
Orbitals are for mathematicians, organic chemistry is for people who like to cook. -Alexander shulgin
-E. Borodin
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micro
bunbun has a gungun



Registered: 05/09/03
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AFAIK, it's debated as much past tryptophan decarboxylase. I think that was from Gewirtz et al. but I don't remember; it would be easy enough to test -- all you'd need is to add glyphosate (Round Up) to the substrate and it should stop producing alkaloids.
I'm not in molbio anymore. I moved to San Francisco and sold out. I've been a software developer for the last decade or so... The pay is a lot better though :3
-------------------- Any research paper or book for free (Avatar is Maxxy, a character by Mizzyam, RIP)
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Coincidentiaoppositorum
deep psychedelic

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Re: Tryptamines in substrate with DPT? [Re: micro]
#22241074 - 09/15/15 12:03 PM (8 years, 4 months ago) |
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Quote:
micro said: AFAIK, it's debated as much past tryptophan decarboxylase. I think that was from Gewirtz et al. but I don't remember; it would be easy enough to test -- all you'd need is to add glyphosate (Round Up) to the substrate and it should stop producing alkaloids.
I'm not in molbio anymore. I moved to San Francisco and sold out. I've been a software developer for the last decade or so... The pay is a lot better though :3
Regardless your a smart person, the psychedelic community needs all the intellegent individuals it can get, even if your contributions are theorys in a notebook or things you publish so who ever can do the most with them will find the information, it would be a waste to not try to share the things you know, specially in a field like psychedelics, where there is no college classes and where misinformation is rampant.
-E. Borodin
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Toadstool5
A Registered Mycophile



Registered: 01/22/15
Posts: 1,359
Loc: The Golden State
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Quote:
glyphosate (Round Up) to the substrate and it should stop producing alkaloids.
On the subject of inhibiting enzymes. Does anyone know what catalyzes the indole-4-monooxygenase for the hydroxylation? I assume it is some cytochrome P450 and was wondering if it can be inhibited without causing other issues with the metabolism?
Any help would be super duper appreciated
-------------------- If you do not know where the mushroom products you are consuming are grown, think twice before eating them. - Paul Stamets AMU Teks Stro's Write Ups
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micro
bunbun has a gungun



Registered: 05/09/03
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Re: Tryptamines in substrate with DPT? [Re: Toadstool5]
#22244957 - 09/16/15 09:14 AM (8 years, 4 months ago) |
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Quote:
Toadstool5 said:
Quote:
glyphosate (Round Up) to the substrate and it should stop producing alkaloids.
On the subject of inhibiting enzymes. Does anyone know what catalyzes the indole-4-monooxygenase for the hydroxylation? I assume it is some cytochrome P450 and was wondering if it can be inhibited without causing other issues with the metabolism?
Any help would be super duper appreciated 
K, now I'm lost.
a.) What context the quote has, and
b.) are we talking about mushroom metabolism or the metabolism of mushrooms (by people) now?
Reason I asked is you mentioned cytochrome P450. I can't imagine you were asking about metabolic pathways in mycelium or you obviously answered your own question. If you're not, well... I really don't know what we're talking about in the first place.
The two kind of contradict :V
Just to clarify: When I hear "cytochrome P450" I generally think "drug interactions."
But, since (I hope) we are talking about mycelial production of alkaloids you answered your own question -- it's an enzyme, which *is* the catalyst.
Maybe I'm missing something.
-------------------- Any research paper or book for free (Avatar is Maxxy, a character by Mizzyam, RIP)
Edited by micro (09/16/15 09:46 AM)
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Toadstool5
A Registered Mycophile



Registered: 01/22/15
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Loc: The Golden State
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Re: Tryptamines in substrate with DPT? [Re: micro]
#22249015 - 09/17/15 12:20 AM (8 years, 4 months ago) |
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In the context of Mushroom metabolism.
As far as I understand things:
There are many different cytochrome p450 enzymes. I can only find information on tryptophan 5-monooxygenase and tryptophan 5-hydroxylation and even less on their inhibition.
I do not know if a tryptamine 4-monooxygenase Cytochrome p450 enzyme exists and if inhibiting it will cause other imbalances with different cytochrome p450 type enzymes within the mycelium.
I assume the 4-hydroxylation of indoles normally involves the same type of cytochrome P450 dependent monooxygenases as the 5-hydroxylation. The problem is, I don't know what can be done to inhibit the 4-monooxygenase and 4-hydroxylase without inhibiting other enzymes and whether or not techniques for the inhibition of 5-monooxygenase and 5-hydroxylase work.
Glyphosate for example, would work very well to inhibit whatever cytochrome p450 type monooxygenase and hydroxylase is causing the reaction but it would also shut down many other cytochrome p450 enzymes
So
1.Is there a specific Cytochrome P450 protein involved in the tryptamine 4-monooxygenase/hydroxylation that anyone knows of?
2. Can it be inhibited using the same methods for the tryptamine 5-monooxygenase/hydroxylation cytochrome p450 proteins?
-------------------- If you do not know where the mushroom products you are consuming are grown, think twice before eating them. - Paul Stamets AMU Teks Stro's Write Ups
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Coincidentiaoppositorum
deep psychedelic

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Re: Tryptamines in substrate with DPT? [Re: Toadstool5]
#22249763 - 09/17/15 07:07 AM (8 years, 4 months ago) |
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Their inhibition? Mono amine oxidase inhibitors should inhibit the molecules metabolism for a moment as the molecules are monoamines...I once took 3g peganum harmala with 3g stropharia cubensis, and it was overwhelmingly intense and long lasting...
Otherwise
www.maps.org/research-archive/w3pb/2002/2002_Passie_22704_1.pdf
http://www.ncbi.nlm.nih.gov/pubmed/14578010
Maybe these will help, I still don't understand fully what your asking about inhibition of metabolism though...
-E. Borodin
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micro
bunbun has a gungun



Registered: 05/09/03
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Glyphosate inhibits the Shikimic Acid Pathway.
I don't know the specifics about the enzymes involved. I'm not sure if it is even known.
-------------------- Any research paper or book for free (Avatar is Maxxy, a character by Mizzyam, RIP)
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Toadstool5
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Registered: 01/22/15
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Loc: The Golden State
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Re: Tryptamines in substrate with DPT? [Re: micro]
#22251799 - 09/17/15 03:36 PM (8 years, 4 months ago) |
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MAOI's inhibit the oxidation of the N-H bonds in amines so I don't believe they would help prevent the C-H oxidation at the 4 position.
Quote:
Glyphosate inhibits the Shikimic Acid Pathway.
I don't know the specifics about the enzymes involved. I'm not sure if it is even known.
Glyphosate would work great but i want the shikimate pathway to work up to the point of tryptamine/NMT/DMT synthesis and then stop it at the tryptamine monooxygenase/hydroxylation so the mushrooms only produce tryptophan/tryptamine/NMT/DMT.
It seems that the exact enzymes for the tryptamine-4-monooxygenase and tryptamine-4-hydroxylase aren't known yet so I will probably use some of the methods used to inhibit the tryptamine-5-monooxygenase and tryptamine-5-hydroxylase and see if psilocybin, psilocin, or baeocystin are still produced.
Do you guys see where i'm getting at here? If you can inhibit the enzyme(s) responsible for the oxidation of tryptamine, NMT, and DMT then in theory DMT can be produced from filamentous fungi without genetic modification. Most sources are biological or GMO.
We need more info on the 4 position mechanism, how it relates to and differs from serotonin and the 5 position mechanisms in particular.
Look at all the speculation over 4-hydroxy-5-methoxydimethyltryptamine and lack of experimental data 
This is one article I am really interested in applying to psilocybian species to prevent the 4-hydroxylization of tryptamine, NMT, and DMT:
Quote:
A Putative Metabolite of Serotonin, Tryptamine-4,5-Dione, Is an Irreversible Inhibitor of Tryptophan Hydroxylase: Possible Relevance to the Serotonergic Neurotoxicity of Methamphetamine
Monika Z. Wrona and Glenn Dryhurst Department of Chemistry and Biochemistry, University of Oklahoma, Norman, Oklahoma 73019
Chemical Research in Toxicology
Vol. 14: Issue. 9: Pages. 1184-1192 Publication Date (Web): August 4, 2001
DOI: 10.1021/tx010037c
Tryptamine-4,5-dione (T-4,5-D) is formed as a result of oxidation of 5-hydroxytryptamine by superoxide (O2-·), nitric oxide (NO·), and peroxynitrite (ONOO-). T-4,5-D rapidly inactivates tryptophan hydroxylase (TPH), derived from rat brain, probably as a result of covalent modification of active site cysteine residues. The activity of TPH exposed to T-4,5-D cannot be restored by anaerobic reduction with dithiothreitol (DTT) and ferrous iron (Fe2+) indicating that the inactivation is irreversible. 7-S-Glutathionyl-tryptamine-4,5-dione, formed by the rapid reaction between T-4,5-D and glutathione, also inhibits TPH but in this case the activity is restored by anaerobic reduction with DTT/Fe2+. The results of this investigation may be relevant to the initial reversible and subsequent irreversible inactivation of TPH evoked by methamphetamine and 3,4-methylenedioxymethamphetamine.
Edited by Toadstool5 (09/17/15 03:53 PM)
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micro
bunbun has a gungun



Registered: 05/09/03
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Loc: Brick City
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Re: Tryptamines in substrate with DPT? [Re: Toadstool5]
#22252141 - 09/17/15 04:57 PM (8 years, 4 months ago) |
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RNAi
edit: that's assuming you have it sequenced
otherwise, maybe look for self-feedback downregulation mechanisms?
-------------------- Any research paper or book for free (Avatar is Maxxy, a character by Mizzyam, RIP)
Edited by micro (09/17/15 05:20 PM)
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Coincidentiaoppositorum
deep psychedelic

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Re: Tryptamines in substrate with DPT? [Re: micro]
#22254615 - 09/18/15 06:03 AM (8 years, 4 months ago) |
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So your trying to inhibit hydroxylation or pbospborlysstion to obtain DMT in your final product?
-E. Borodin
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micro
bunbun has a gungun



Registered: 05/09/03
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4-OH
At least, that's what I got out of it.
I wasn't even aware anything was sequenced yet, until your post.
Then again, I've been out of the loop for like a decade or so 
edit: dyslexia *rolleyes*
-------------------- Any research paper or book for free (Avatar is Maxxy, a character by Mizzyam, RIP)
Edited by micro (09/18/15 10:12 AM)
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Coincidentiaoppositorum
deep psychedelic

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Last seen: 8 years, 4 months
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Re: Tryptamines in substrate with DPT? [Re: micro]
#22255612 - 09/18/15 10:37 AM (8 years, 4 months ago) |
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4-HO-DMT is psilocin...
This is why this is all confusing to me, it seems like several different things have been jumbled together in a way to make it as unclear as possible, I'm interested in the topic here, but I'm having trouble trying to decipher your intent.....meaning I generally know the processes and enzymes here, but its hard for me to tell what exactly your trying to say.
There's a grid for the biosynthetic pathway of psilocybin because after tryptophan decarboxylation its unclear what pathways the fungi are using, there are a few diffferent possibilities...
It sounded to me like you were asking if by inhibiting hydroxylation and Phosphorylation of the dimethyltryptamine if you could yeild it in the fruiting bodies...
But I got somewhat lost...
-E. Borodin
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