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Mushrooms, Mycology and Psychedelics > Advanced Mycology
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Re: Adding tryptamines into Mushroom Substrate [Re: leftover_crack]
    #11171032 -

leftover_crack said:
if i recall correctly the mushroom only produces the added substituted version as substituted tryptamines inhibit the tryptophan decarboxylase hence DET prevents tryptamine and hence psilocin from forming leaving only 4-ho-DET to be produced.


Do you have a citation for that?  I've never read about feedback inhibition for TDC anywhere and would be interested.


-FF

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Re: Adding tryptamines into Mushroom Substrate [Re: fastfred]
    #11211589 -

after rereading some papers i think i may have mixed up papers because i was reading one about tryptophan feedback loop in Catharanthus roseus but that was due to tryptophan production inhibition. however in the gartz DET > 4-ho-DET paper it does say no additional psilocin or psilocybin was detected. whether that means in addition to the 4-ho-det or in addition to normal levels i'm not sure. its not worded very well.

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Re: Adding tryptamines into Mushroom Substrate [Re: leftover_crack]
    #11212075 -

I don't see how it would be feasible for mycellium to carry the digested drugs to the fruit bodies. I don't see how an organism could be tricked into carrying something non-nutritious to the actual mushroom. I wouldn't be surprised if it was still in the substrate though.

I think that would be the real way to test would be to cook up a cake the same way as always, pc, then inject drug/chemical-solution into it in a concentrated area/side, then inject the spore solution or lc allow full colonization then do some kind of test for the drug. I'd imagine doing something with melotonin would be a good cheap way to go about doing this.

If the original chem was still there that would mean the myc did nothing at all(pretty expected).
if it turned into something else(booya)
if it was eaten and turned into nothing then the mycellium just eats is(ouch).

I also don't see why it would eat a drug, but then again Myc will eat aluminum..
If anyone has the skills/tools to look for changes in chemicals I'll try it with Oyster Mycellium and send it off via mail. Or would it only work for a psilocybin producer? I want to put these ideas to rest for the sake of science...

Edited by smaerd (10/08/09 11:42 PM)

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Re: Adding tryptamines into Mushroom Substrate [Re: smaerd]
    #11219507 -

smaerd said:
I also don't see why it would eat a drug, but then again Myc will eat aluminum..


This hasn't been established. It's possible the metabolites are corroding the aluminum and the myc is not absorbing it back up.


--------------------
The above post is fictional, hypothetical, or downright nonsensical.


:moon:  :moon:  :moon:  :moon:    :moon:

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Re: Adding tryptamines into Mushroom Substrate [Re: anonjon]
    #11276284 -

This is silly...I started a thread like this nearly 2 years ago and got the same results, a lot of talk and nothing to show. I was interested but not so much anymore, and I didn't have the resources back then (I still don't). The problem is that even though its an awesome thought experiment and would be cool as hell if it worked, its not worth the extra time and money to make this a standard practice in mushroom cultivation.

But I love all the thought put into it, its very interesting to discuss the possibilities :grin:

peace
agmotes165


--------------------
The universe is under no obligation to make sense to you - NDT

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Re: Adding tryptamines into Mushroom Substrate [Re: Jean-Luc Picard]
    #11278120 -

Usually the mods close these threads for the reasons you describe above.


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The above post is fictional, hypothetical, or downright nonsensical.


:moon:  :moon:  :moon:  :moon:    :moon:

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Re: Adding tryptamines into Mushroom Substrate [Re: smaerd]
    #11281058 -

smaerd said:
I also don't see why it would eat a drug, but then again Myc will eat aluminum..
.


I think your mixing vernacular with science terms and messing up a bit...

First, myc don't "eat" the way you and I do...they secrete enzymes that break down the substrate, and have transporter proteins (I assume they don't feed solely off osmosis...) that recognize a compound, grab it and take it inside for further pathways.

The pathway is somewhat known starting from tryptophan, which is an amino acid commonly found in nature (especially decaying manure, I would imagine....). If we modify that substrate slightly, according to the original research, the myc won't know the difference (since proteins don't have brains), grab it and push it through the pathway.

I a 90% certain 4-AcO won't work because that ester group is polarized enough where tryptamine/tryptophan is not. 2c-e isn't even the right ball park...its a phenethylamine, not a tryptamine.

I am willing to believe the research presented on it since it seems that mush. grown on different substrates have different potencies (or so has been the going myth), and the best is supposed to be Hpoo(also, according to myth...none of this has been assayed AFAIK)....Hpoo seems to me like it'd be a hefty source of indole, tryptamine, tryptophan and more importantly, the gut flora that would happily connect the dots there.

Just my 0.02

-jon


--------------------
--------------------------------------------------------------------------------------------------------

Microbiologist turned mycologist, but still amateur enough to enjoy the occasional surprise...

My Trade List

Terry M said:
Looking for rules that other people use successfully. Sterile procedure is just a set of rules that work.

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Re: Adding tryptamines into Mushroom Substrate [Re: tr8orjohn]
    #11281187 -

Ooo Thanks for the knowledge. It's always nice to hear a scientific mind at work.

In which case Sun-flower seeds may be a good test substrate for tryptophan. It has 3x the amount compared to white wheat flour(according to wiki). As for the other chemicals in question I'm not sure.

If you have any ideas for an experiment please post it up and explain it in lamends terms so someone on here could test it. I want these kinds of threads to stop popping up unless the are success stories of course.

Edited by smaerd (09/13/10 07:15 PM)

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Re: Adding tryptamines into Mushroom Substrate [Re: smaerd]
    #11282156 -

Well, L-Tryptophan is available online as a dietary supplement/sleep aid.

Seems like a good experiment would be to use an isolate culture to inoculate several jars of BRF PF tek jars as a control, and knock up several BRF PF tek jars with various levels of added L-Tryptophan with the same isolate culture as the control jars, grow them out and test for potency.

Easier said than done (I dont have a way to accurately test for psilocybin/psilocin concentration). But it seems like a legit setup.

Feel free to tear my setup to shreds if it sucks haha.

peace
agmotes165


--------------------
The universe is under no obligation to make sense to you - NDT

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Re: Adding tryptamines into Mushroom Substrate [Re: Jean-Luc Picard]
    #11284301 -

@smaerd: if you poke around the archives you'll find 2 things of interest:

Tryptophan utilization is limited by the enzyme that breaks it down into tryptamine, which itself inhibits that enzyme.  Adding tryptophan probably won't help.

A paper or two on radioisotopes of cybin precursors and their tested uptake into mycelia.  NMT is the one with the highest uptake by a wide margin.  That's probably your best bet.

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Re: Adding tryptamines into Mushroom Substrate [Re: urbanfarmer]
    #11284514 -

Quote:
Tryptophan utilization is limited by the enzyme that breaks it down into tryptamine, which itself inhibits that enzyme.  Adding tryptophan probably won't help.


Find a citation for that.  I've heard it a couple times now with nothing to back it up.

Quote:
Seems like a good experiment would be to use an isolate culture to inoculate several jars of BRF PF tek jars as a control, and knock up several BRF PF tek jars with various levels of added L-Tryptophan with the same isolate culture as the control jars, grow them out and test for potency.



It's been tried at least a couple times now.  Tryptophan is not taken up by the myc.








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Re: Adding tryptamines into Mushroom Substrate [Re: fastfred]
    #11305648 -

Good stuff fastfred.  I haven't (until recently) done extensive reading/research into topic of tryptamine supplmentation, but searching your name via the search function has proved nothing less than informative and very worthwhile - i can see you've done your homework.  I was about to give up hope after reading through all of the BS posts on here provided by individuals who are either misinformed or feel they have sufficient knowledge to jump to [uncited] conclusions that have little to no scientific backing and have not been replicated in the lab.

fastfred said:
1. Tryptophan is too highly regulated to cause much if any increase in psiloc(yb)in.

4. Tryptamine supplementation increases the psilocin, but then the phosphorylation to psilocybin becomes the limiting step.

5. Phosphate content of the substrate could be a way to regulate the psilocin/psilocybin ratio. Otherwise phosphorylase activity becomes the limiting step after tryptamine and/or TDC have been increased.


1.  This was my first thought when reading though all of the posts on tryptophan and why I was able to mentally debunk this theory.  Reading this was very refreshing to me.

4.  I don't know if this has been explained, but I don't fully understand why an increase in psilocin levels causes a decrease in phosphorylase activity.  Does anybody have an answer, or is this just an observed/proposed trend?

5.  Phosphate supplementation increasing phosphorylase activity is a pretty thought, however I feel like this theory implies that phosphate levels in the substrate are already at a deficit - and also requires that phosphate carrier molecules associated with phosphorylase enzymes will indeed uptake phosphate from secondary sources and incorporate them into the psilocybin pathway.


--------------------
"Sucralose (Splenda) is a chlorocarbon.
it's in the same family of chemicals as DDT and PCBs.
Don't worry, it’s not DDT, it’s “DDT-Lite!”

Edited by staratsx (10/23/09 11:57 AM)

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Re: Adding tryptamines into Mushroom Substrate [Re: fastfred]
    #11309247 -

I just came all over my laptop. Thanks for the papers


--------------------
http://flickr.com/photos/mycography - A collection of my fungal photography

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Re: Adding tryptamines into Mushroom Substrate [Re: kydelic]
    #11314788 -

kydelic said:
I just came all over my laptop. Thanks for the papers


happens sometimes...i love scientific articles and sites that back up info...it gives me a warm and fuzzy feeling...

and staratsx...i believe for 4 he was saying that although psilocin production and concentration increases, you reach a point where due to the availability of phosphate and the "fixed" reaction rate of the enzyme, that the conversion of psilocin to psilocybin would be the limiting factor to long-term potency increase, since psilocin doesn't last long in drying and storage. So while fresh fruits may be more potent, the older dry product may not be that much more potent.

feel free to correct me if im dumb haha

peace
agmotes165


--------------------
The universe is under no obligation to make sense to you - NDT

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Re: Adding tryptamines into Mushroom Substrate [Re: Jean-Luc Picard]
    #11315022 -

Quote:
agmotes165 said: ....the conversion of psilocin to psilocybin would be the limiting factor to long-term potency increase....


yes this would also be true, but I'm not sure that's what he was referring to.  I remember reading another post by someone (which was cited), proving that the increase in psilocin levels achieved by tryptamine supplementation brought on a dramatic decrease in psilocybin formation, which was only found in trace amounts. 

dispite the fact that the last step of the pathway is the phosphorylation of psilocin, I feel like we are jumping to conclusions when we say that phosphate is at a shortage (although it seems it may be a logical conclusion).  one thing holds true though...phosphorylation to psilocybin is indeed the limiting step, but why?


--------------------
"Sucralose (Splenda) is a chlorocarbon.
it's in the same family of chemicals as DDT and PCBs.
Don't worry, it’s not DDT, it’s “DDT-Lite!”

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Re: Adding tryptamines into Mushroom Substrate [Re: staratsx]
    #11317280 -

There are 2 limiting steps: decarboxylization (I can't spell) of tryptophan into tryptamine,
phosp of cin to cybin.

In terms of production, I'd call the first the primary limiting step.  You could always do an extraction with phosphoric acid to get around the second step.

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Re: Adding tryptamines into Mushroom Substrate [Re: urbanfarmer]
    #11317901 -

urbanfarmer said:
There are 2 limiting steps: decarboxylization (I can't spell) of tryptophan into tryptamine,
phosp of cin to cybin.

In terms of production, I'd call the first the primary limiting step....


Well the first step is certainly limiting if we're talking about tryptophan, and i would consider it limiting enough to dismiss the practice of tryptophan supplmentation all together (just decarboxylate yourself). 


Quote:
urbanfarmer said:You could always do an extraction with phosphoric acid to get around the second step.


I'm sry i don't really know what you mean by this.  are you referring to the addition of phosphate to the growing medium, or actually running a psilocin extraction and treating it with phosphoric acid?  The latter of which would not work, as the reaction (based on my knowledge) is not a favorable one, as the molecule is incapable of nucleophilic attack w/o the help of a kinase


--------------------
"Sucralose (Splenda) is a chlorocarbon.
it's in the same family of chemicals as DDT and PCBs.
Don't worry, it’s not DDT, it’s “DDT-Lite!”

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Re: Adding tryptamines into Mushroom Substrate [Re: staratsx]
    #11323035 -

Do you have numbers on the phosphate/phenol ester formation?  Not having those, I'd assume that this would be a dehydration bond like any other.

re: the trypt[ophan/amine] question, there's still the uptake issue to contend with.

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Re: Adding tryptamines into Mushroom Substrate [Re: Doc_T]
    #11349145 -

Doc_T said:
Look around on eBay, maybe.


i found my gc on the side of the road


--------------------
"Sucralose (Splenda) is a chlorocarbon.
it's in the same family of chemicals as DDT and PCBs.
Don't worry, it’s not DDT, it’s “DDT-Lite!”

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Re: Adding tryptamines into Mushroom Substrate [Re: staratsx]
    #11358616 -

Howdy,




Attached is a pic of an in-vitro fruiting that I believe was a result of adding sterilized "reed canary grass" seed to an inoculated plate contains a culture of P "???" These spores came to me as "super K samui" and has proven to be a finicky strain to work with. No matter what the agar substrate, the preferred rizomorphic growth patterns were hard to come by, often degrading into airier cottony (cepatose) patterns. Definitely not a cube, and highly active.

And then one day the reed canary grass became ripe, so I collected some sample grain dried and sterilized and added whole seed right on to the plate. The cultures were previously inoculated on 60MM PMEA dishes. Of 10 dishes only 2 received RCG supplement.

Both cultures that were augmented grew until contact was made with the grain, stopped (presumably to develop a new digestive enzyme) and then captured the grain with vigor. A fruiting occurred so large ( In both cultures) that the stem base of the mushie burst the Parafilm securing the culture!! The fruit persisted for several weeks and grew to the maturity of SPOREULATING in culture. The mycelium and mushrooms bruised dark blue withing seconds of scalpel contact and were presumed to be highly active.

The non augmented cultures showed no deviation in growth patterns. All cultures were kept in the same incubation.

Paul stametes makes an off hand comment in Psilo of the World that researchers had added DMT to substrate to make especially active fruits, but never stops to explain the how or why. The book is partially response-able for the idea of the experiment, the other part coming from the observations that certain species of  high potency strains (e.g. P. Azurecens) are associated with DMT containing dune grasses in nature (sleepy grass), leading me to believe that those grasses were feeding the mushies from dead roots (where most of the DMT is contained) suggesting that the association was something more than habitat, highly perhaps even a deep symbiosis.


I have only achieved arc-typal growth patterns for this species using a deviation of this technique. I consider the growth patten in the picture to be prime for this species, presumetaion being supported by a 2" sporeulating fruit accompanied by a smaller twin fruit and a primordia setting in the lower right of the pic. Sub cultured plates showed good growth over a couple generations, then degraded back towards cottony ariel growth with little rizomorphic patterns.

I do plan to work some more with this next year, perhaps starting a 10% substrate augmentation with the reed canary grass. Strict controls in place of course.


If any of You have ever run across "Super K Samui" and know exactly what that is please drop me a line. That is one of my great unanswered questions.

I certainly thing that this idea could bear "fruit"

Mycoelf


--------------------
Mycoelf

Sterility is a process that can be likened unto infinity, which is a long walk, the closer to the end you start before beginning, the more achievable  the goal of infinity becomes.  Remember, cleanliness in next to goddessness

:aliendance::aliendance::wicca::aliendance::aliendance::pipesmoke:

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