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OfflineDucky
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Re: Tryptophan Substrate? [Re: Ducky]
    #5642016 - 05/17/06 01:58 PM (17 years, 10 months ago)

Ok so while I'm having my lunchtime soup here and fantasizing about doping up my next substrate, would adding something like phenylalanine (probably the dextro- DPA) to substrate make a mushroom more "speedy"? Afterall, decarboxylation of phenylalanine results in phenylethylamine, your brain's endogenous amphetamine, also the backbone of many amphet analogues. PIHKAL anyone?


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"Whoa! ...did you just see that too?"

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InvisibleGio
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Re: Tryptophan Substrate? [Re: Ducky]
    #5643332 - 05/17/06 07:30 PM (17 years, 10 months ago)

Well, I think it's a little trickier than that. Removing the -COOH from a Phe would probably require a different enzyme (i.e. phenlyalanine decarboxylase). And despite what I proposed in my last post, I still think adding more psilo precursors (e.g. tryptophan, tryptamine) might be contradictive. Trp decarboxylase is inhibited by its product, tryptamine, probably because too much of the amine will alter the intracellular pH. A slight change in pH can completely ruin many enzymes, so there are mechanisms put in place to maintain balance (e.g. feedback inhibition); Elemicin mentioned homeostasis. Anyway, psilo products are probably just intermediates like tryptamine.

Edited by Gio (05/18/06 01:28 AM)

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OfflineDucky
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Re: Tryptophan Substrate? [Re: Gio]
    #5643460 - 05/17/06 08:02 PM (17 years, 10 months ago)

I totally hear ya. Just a side note..as far as 5htp is concerned, yes with the possibility of it being more of a precursor to Norbaeocystin, well anyways, a few jars with a moderately high 5htp content in the substrate are just exploding in growth, much faster than the identical ones started two weeks earlier, same strain, same syringe. Interesting to see how anything is affected, aside from how growth seems to be so far.

If I understand correctly, decarboxylation is similar if not otherwise to fermenting. So, in lamen's terms, fermented tryptophan would tend to yield tryptamine. What about acetylation now? I'm not too hot with chem, but if one were say to add enough base to some acetic acid to neutralize the pH, would there maybe be anything there to aid in psilocybin synthesis?


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Invisiblefastfred
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Re: Tryptophan Substrate? [Re: Ducky]
    #5656906 - 05/21/06 04:00 PM (17 years, 10 months ago)

Quote:

Ducky said:
More reading on it happened...here's this link again..
http://www.shroomery.org/index.php/par/24373




Interesting article. However, most of the information isn't that relevant to psilocybin/psilocin. The only important steps are from tryptophan to tryptamine and from tryptamine to psilocybin. These are the important steps and unfortunately they are the ones we know little about. From reading this you might get the idea that we know a lot about the psilocybin biosynthetic pathway, but the fact is we don't.

Every living organism produces tryptophan, it's one of the essential amino acids and is required for life. Some biosynethetic pathway produces tryptophan in all life, so it isn't specific to psilocybin production by any stretch of the imagination.

The information on tryptophan decarboxylase inhibition is interesting, but there is no reference for it. It looks like it comes from an interesting paper, so it's a shame that it's been reduced to a simple table.

The tryptophan decarboxylase information is great, but the rest of the article can be reduced to:
Tryptophan =(Tryptophan decarboxylase)=> Tryptamine =(???)=> (???) =(???)=> Psilocybin

This cuts out the irrelevant information and more accurately represents our actual knowledge of the biosynthetic pathway (or lack thereof).


-FF


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It drinks the alcohol and abstains from the weed or else it gets the hose again. -Chemy

The difference between the substances doesn't matter. This is a war on consciousness, on our right to the very essence of what we are. With no control over that, we have no need to speak of freedom or a free society. -fireseed

"If we are going to have a war on marijuana, the least we can do is pull the sick and the dying off the battlefield." -Neal Levine (MPP)

I find the whole "my drug should be legal but yours should be illegal" mindset disgusting and hypocritical. It's what George Bush and company do when they drink a cocktail and debate the best way to imprison marijuana users. -Diploid

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InvisibleGio
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Re: Tryptophan Substrate? [Re: Elemicin]
    #5682286 - 05/28/06 01:14 AM (17 years, 10 months ago)

Elemicin said:
"Besides pH in the substrate should always be slightly basic (7.2 is what I use) and this keeps contamns down."

Wow...after a few hours study, I think I see your point. I re-read your posts too and had some insights I wanted to share for criticism.

It looks like a basic pH might play an indirect role in psilocin's phosphorylation: first, tryptamine's indole-4 would be easier to hydrate. Then, the extra mass in psilocin (perhaps also complimented by some force of attraction) sinks the molecule closer to the enzyme's ATP-binding domain(s), where esterfication of psilocin is a consequence. The PO4 in psilocybin raises the molecular mass dramatically, so its less mobile, and it exponentially aggregates, which reduces the enzyme's capacity and activity. Eventually, the region where psilocin esterfication occurs becomes blocked by a critical mass of psilocybin.

This gives me the image of a pipe or cavity gradually being filled and clogged (feedback inhibition). Enriching the sub with precursors will only make this happen sooner. I guess either increasing the number of these channels and/or their capacity would result in some crazy (maybe lethal) increase in potency, but that's GE.

Okay, can someone show me the loose-ends?

EDIT: I meant DMT when I wrote tryptamine.

Edited by Gio (05/28/06 01:39 AM)

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InvisibleGio
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Re: Tryptophan Substrate? [Re: Gio]
    #5682434 - 05/28/06 02:24 AM (17 years, 10 months ago)

This probably belongs in a new topic:

I was smoking and wondering about the structure of the enzyme( (or complex).  I'm imagining a funnel-like structure, where the sub is processed along a track of cofactors/coenzymes in a downward spiral fashion, towards the narrow stem from which PO4 is introduced...kind of like a continuous flushing of something down a toilet where a little is behind everytime (until it clogs). 

For the moment, I'll nickname this structure "psilocybin-conicalase" (for its conical shape), or "psiloconicase."  I also wonder how the internal volume is distributed in psiloconicase and how it's controlled...

Maybe certain scaffold proteins are attatched to the enzyme's exterior and receive signals on when to flex (via depolymerization) and relax (via polymerization)... i.e. pulling on the surface and stretching its  dimensions.  How would changing psiloconicase dimensions affect its activity (i.e. more depth vs. more width)?  And would influencing these scaffold parts be an approach for increased psilocybin production?

Wow...I go 7 days without smoking, and tonight I'm baked.  Kick ass!

EDIT:  More words :smile:

Edited by Gio (05/28/06 03:18 AM)

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OfflineAbortfungi
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Re: Tryptophan Substrate? [Re: Gio]
    #5685743 - 05/29/06 07:31 AM (17 years, 9 months ago)

This might be off topic, but I've managed to get some L-tryptofan containing pills. Now, is there any evidence how these might play with one's mind if consumed with p.cubensis? These pills also contains: L-glutamine, L-tyrosine, magnesium and vitamin B6.
These pills are a extra nutrient product for insomnia or for one to get some sleep. Bought from natures meds shop. Does anyone have any information how I could use this source of tryptofan to anything fun?

Sorry bout my english...


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OfflineDucky
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Re: Tryptophan Substrate? [Re: Abortfungi]
    #5734647 - 06/10/06 03:43 PM (17 years, 9 months ago)

Using a standard BRF substrate as just an example of ratios between verm, water, and nutrients, how about this:

"Bee Pollen has high concentrations of the B vitamin complex, and also contains Vitamins A, C, D, and E. Its composition is: approximately 35% protein, 55% carbohydrates, 2% fatty acids, and 3% minerals and vitamins"

"Rice, brown (long-grain, cooked), 1 cup (195g)
Calories: 216
Protein: 5g
Carbohydrate: 44.7g
Total Fat: 1.75g
Fiber: 3.5g"

Screw the BRF. One interesting thing is that pollen has those B vitamins - they are essential cofactors for metabolism of many many amino acids and production of many neurotransmitters, at least in primates. I'd imagine that the metabolic pathways are much more similar in fungus than we'd imagine.

I can imagine a substrate of approx (by volume):
40 units vermiculite (coarse grade)
20 units water
15 units tryptophan
2-3 units bee pollen
2 units whey protein
1 unit trace minerals

I don't know what kind of pH a mixture like this would reveal, but a simple fix would of course be to add some crushed oyster shell of whatever else you'd prefer.

Another similar mixture would be possibly to substitute half of the tryptophan for melatonin, but it would probably be a totally different result anyways. I know I keep coming back to the melatonin idea, but I really think there's something there. I have NO clue how P. Cubes metabolize it, if they even can (I really don't think they use it or produce it naturally), but I think it might be an easier possible way to end up with something we want the fungus to produce from a simple additive or combinations of them.

I forgot to add in a bit of flax seed flour/meal to the recipe, maybe 3 parts for fatty acids.


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"Whoa! ...did you just see that too?"

Edited by Ducky (06/10/06 03:52 PM)

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InvisibleZen Peddler
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Re: Tryptophan Substrate? [Re: fastfred]
    #5936346 - 08/06/06 07:29 AM (17 years, 7 months ago)

'Well, it looks like there are at least several experienced cultivators who have grown on plenty of substrates, with varying levels of tryptophan and nobody can seem to agree if it affects potency or not!'

Actually, five years ago or so growers like Major Millet, Uma Guma, and a few others were always suggesting that millet tended to grow more potent cubensis purely from bioassay. This was discussed at that time (around 2001 if i remember) and as nearly everyone who tried it at the time agreed, it was taken as fact.
This lead to a review of why millet yielded more potent cubensis and larger yields and fruitbodies (again only measured by bioassay) and the again the only significant variation found between brown rice flour and millet was their tryptophan contents.
Not exactly rocket science, but since those on the old drool were sound individuals the finding were assumed to be correct.
My assumption coming from the fact that I have never found the same content of brown rice substrate to yield mushrooms of the same calibre as millet, rye or other grains - millet being the best (hence major millet's choice of name).


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InvisibleZen Peddler
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Re: Tryptophan Substrate? [Re: Zen Peddler]
    #5936347 - 08/06/06 07:31 AM (17 years, 7 months ago)

And measured by the fact that ultimately all precursors of psilocin are ultimately made from tryptophan


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Offlineimjrod
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Re: Tryptophan Substrate? [Re: Ducky]
    #20787905 - 11/03/14 12:24 PM (9 years, 4 months ago)

Just to spread encouragement to others, I've been doing this for many years now and it does work. D-Tryptophan makes a bigger difference in slow growing "strains" or those with slow growth periods like Peurto Rican. Slow strains given tryptophan will be more than twice as strong. It's nice to only have to eat half as much shrooms.

But, don't get too excited. The only real difference is I don't gotta eat as much.

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InvisibleThe Lightning
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Re: Tryptophan Substrate? [Re: imjrod]
    #20835123 - 11/14/14 05:46 AM (9 years, 4 months ago)

Quote:

My take is this: tryptophan is a precursor to 5-HT (5-hydroxytryptamine / serotonin). Perhaps this will boost tryptamine levels and increase potency?





Quote:

MarkostheGnostic said:
Hey Bud -
<br>According to CB Gold in Psychedelic Monographs and Essays, #6 (T. Lyttle, ed.) in the article 'The Mushroom Entheogen: Part 3a - Nutritional Influences on Growth and Psychedelic Biosynthesis,' page194, he says: "Tryptophan is an important amino acid in the synthesis of psilocybin. One may rightly think that adding this amino acid to the media may increase the final psilocybin production. Apparently this is not the case....Analysis of carbon-14 uptake by p. cubensis in liquid culture showed that tryptophan is not directly used by the mushroom to produce psilocybin and the addition of which will not increase the total amount of psilocybin." Apparently, production must be approached at a more fundamental level. "The addition of dextrose yielded the greatest benefit, followed by TMI (trace minerals) and lastly ammonium succinate-glycine. In combined form, the TMI/dextrose was significantly better in growth and biosynthesis than either supplement singularly (p.193)." The TMI is a product from a company called Trace Minerals, Inc., and comes from clay from the ancient Columbia riverbed in Oregon, and contains "trace amounts of just about every metallic element." I've personally used crushed GNC mineral tabs added to the water for the PF type cakes. It's all so variable, and I'm not a biochemist, but it may well have boosted potency.<br><br>









Also, an added hypothesis from a different thread:


Quote:

If you really want to increase psilocybin content, add Diammonium Phosphate and non fat powdered milk to the substrate.  Diammonium Phosphate has the phosphorus needed to make psilocybin, and and non fat powdered milk has the tryptamines.



Edited by The Lightning (11/14/14 06:12 AM)

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OfflineRogerRabbitM
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Re: Tryptophan Substrate? [Re: shymanta] * 1
    #20850057 - 11/14/14 05:46 AM (9 years, 4 months ago)

This thread has been closed.

Reason:
Too many years of this thread spouting opinion as fact with zero supporting evidence.

This is unworthy of advanced mycology.

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