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Re: MIMOSA BARK IN MAiLBOX!!!!! [Re: gray1]
    #331616 -

I guess my basic feeling is not so much favoring one precursor over another, but simply illustrating that the further you are from the end of the path, the more work you must do to take care that you also have an excess of all building blocks from that point on, plus enough to take care of the fungus, as it is doing more work than normal.

This might require more work than it is worth. I don't mean overall. I think these experiments should be attempted (UNDER CONDITIONS THAT DON'T VIOLATE ANY LAWS). I just mean that it might be more work to try and use tryptophan (even for an extreme amature) than to try and obtain, extract, and prepare DMT as a precursor.

I speculate that early atempts at adding tryptamine, which resulted in gnarled stems, and only excess psilocin suffered mainly from lack of this sort of thinking. The fungus was probably doing everything it could to get rid of this excess tryptamine, at expense of its own healthy functioning, and without the benifit of excess phosphate to create psilocybin.

I also want to thank you all for participating in what I feel is one of the most productive threads in a long time. And we really haven't accomplished anything yet! But we are straightening out a lot of muddled thinking on the subject, and I think we are close to a reasonably easy approach to boosting the levels of the desired alkaloids.

OPI


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Re: MIMOSA BARK IN MAiLBOX!!!!! [Re: Opi]
    #331662 -

"I just mean that it might be more work to try and use tryptophan (even for an extreme amature) than to try and obtain, extract, and prepare DMT as a precursor. "


again, i feel that i need to stress that DMT is NOT a good substrate addition
although it is an intermediate in the biological process, studies have shown that it is not incorporated into the organism from the substrate. if it doesn't get into the cells it can't be a part of any biosynthetic pathways, regardless of how good a potentiator it may or may not be. period.
this is why i continue to suggest tryptophan as the only realistic substrate addition for those who are so inclined to carry out experiments. it is readilly availiable, of reasonable expense, and can be incorporated into the organism.

whether the addition of tryptophan will give rise to any noticable difference (increase) in potency and psilocin/psilocybin remains to be seen, and even so will be highly subjective.

also, i am not trying to suggest that the fungi should do "more work than normal" or function abnormally,
just attempting to creat a theoretical optimal environment for the NATURAL function of the organism to it's full inherrent capabilities.

gray1

additionally, i don't see how adding tryptophan to the substrate during regular substrate preparation is more work than growing/ordering, extracting and adding DMT to the substrate, in fact, the reverse seems true to me. i think that any amature could pop open a few capsules and mix it inwith the vermiculite/brf...

Edited by gray1 on 06/01/01 04:06 PM.


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Re: MIMOSA BARK IN MAiLBOX!!!!! [Re: gray1]
    #331790 -

In the "Tryptophan" post, also in this forum, it is stated that primrose seeds and sunflower seeds carry an abundance of biologically produced tryptophan more than any other organic source.

If this info. is accurate than why not just add a little crushed primrose seed or sunflower seed to brf/verm or birdseed substrates.  This help boost levels of alkaloids, but still provide other nutrients needed by the fungus, perhaps avoiding the gnarled fruits.

4 out of 5 inmates agree.  The FSR is better than being raped by your overweight roomate!;)
Support the FSR.


--------------------
Vote Jonnyshaggs in the next election for GOD...Its the responsible choice

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Re: MIMOSA BARK IN MAiLBOX!!!!! [Re: gray1]
    #332198 -

I guess you only read half of what people say... I won't explain why it is less work in this post, as I did more than an adaquate job in my last post.

And actually DMT works fine as a substrate addition.
"studies have shown that it is not incorporated into the organism from the substrate"

Just plain not true. There is a difference between not AS effective, and NOT effective.

It HAS been shown to increase psilo levels, just is not AS effectivly as tryptamine (probably due to solubility).

And if you are elevating the levels of precursors above what you will ever find in nature, I would hesitate to call it natural. It certainly is more work for the fungus than it would encounter in nature..

I know this is something you are investing a lot of thought into, but please don't get defensive over ideas.


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Re: MIMOSA BARK IN MAiLBOX!!!!! [Re: Opi]
    #332199 -

OK.. I can't resist RE-explaining myself...
Tryptophan is plain NOT AN OPTION. By itself it does not work. That HAS been demonstrated. You might be able to "WITH A LOT OF WORK" get it happen. But you would have to take so many things into consideration (i.e. Every step of the biosynthetic process from tryptophan to psilo) to get it to happen, that it would be harder to accomplish than using DMT. And the organism would have a chance to limit you at SEVERAL steps instead of just at the final one.

OPI


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Re: MIMOSA BARK IN MAiLBOX!!!!! [Re: Opi]
    #332202 -

Damn why couldn't I just put this all in one post 8^)

I speculate that the specific reason tryptophan doesn't seem to elevate alkaloid levels is that the mechanism is already functioning at a level where it is converting as much tryptophan into tryptamine, as is going to happen. Maybe I am wrong about being limited at every step. Maybe this is the only limiting step the fungus happens to have, which would make this a little easier. I certainly hope so.


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Re: MIMOSA BARK IN MAiLBOX!!!!! [Re: Opi]
    #333327 -

just a little question here.  If the fungus is limited with tryptophan than it may just as well be limited with tryptamine and DMT and psilo's. 
Maybe we should all just sit down and ask the fungus what it wants.

4 out of 5 inmates agree.  The FSR is better than being raped by your overweight roomate!;)
Support the FSR.


--------------------
Vote Jonnyshaggs in the next election for GOD...Its the responsible choice

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Re: MIMOSA BARK IN MAiLBOX!!!!! [Re: Opi]
    #333875 -

opi:
you say that DMT is proven to work: where is the scientific proof/literature behind this statement? please post a link or reference.

DMT IS NOT INCORPORATED INTO THE ORGANISM
go back and read through this thread again (or more likely for the first time) and you will see the literature reference to this that used a radiolabel incorporation study. if you give me your email i'll attach the paper as a pdf. (let me remind you again of what i have previously stated, in theory, i think that increased in vivo levels of DMT would cause an increase in end product, but simply adding DMT to the substrate will not increase in vivo levels, therefore no increase in end product will be seen.)

you state that adding tryp alone won't work, while i concede that this may be the true, possibly because the substrates commonly used are already supplying all of the needed tryptophan, but you seem to say so definatively.
please reference to what makes you say this.

i consider supplementing to the full nutritional requirements of the mushroom natural. who is to say that the natural environments in which these mushrooms grow don't provide optimal conditions, including nutritional requirements? and why shouldn't we strive to provide equally optimal environments in our artifical process, but do so as naturally as possible?
obviously the difficulty here is twofold:
1. we don't don't know the precise nutritional value of the substrates used
2. we don't know the true nutritional requirements of the organism.
for humans, don't you think that getting the full RDA ov every vitamin and mineral, correct mg/kg protein, correct carbos, fatty acids, etc... is natural? with proper diet it certainly can be achieved, if you don't think that taking vitamins is natural.
again, through supplementation, i only suggest that we try to create the optimal environment for the organism to grow, and let it do it's thing concerning the production of indole alkaloids.




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Re: MIMOSA BARK IN MAiLBOX!!!!! [Re: gray1]
    #333879 -

"I guess my basic feeling is not so much favoring one precursor over another, but simply illustrating that the further you are from the end of the path, the more work you must do to take care that you also have an excess of all building blocks from that point on, plus enough to take care of the fungus, as it is doing more work than normal.

This might require more work than it is worth. I don't mean overall. I think these experiments should be attempted (UNDER CONDITIONS THAT DON'T VIOLATE ANY LAWS). I just mean that it might be more work to try and use tryptophan (even for an extreme amature) than to try and obtain, extract, and prepare DMT as a precursor."

you appear to be speaking about two different "works" here:
the work of the mushroom that is escalated because of supplying more precursor
and the work of the cultivator, supplying more precursor.

my response,

additionally, i don't see how adding tryptophan to the substrate during regular substrate preparation is more work than growing/ordering, extracting and adding DMT to the substrate, in fact, the reverse seems true to me. i think that any amature could pop open a few capsules and mix it inwith the vermiculite/brf...

stands true. the excess work you are refering to is the work of the fungus itself, obviously the cultivator is not responsible for this work.





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Re: MIMOSA BARK IN MAiLBOX!!!!! [Re: gray1]
    #333921 -

bleuboxo:
how's the experiment coming?

gray1


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Re: MIMOSA BARK IN MAiLBOX!!!!! [Re: gray1]
    #336184 -

Auto-Summary
The post discusses the potential of using tryptophan to increase psilocybin content in Psilocybe cubensis mushrooms. The author believes that Gartz, Shulgin, and others were incorrect in recommending this method. Instead, the author suggests extracting DMT and exposing it to the fungi as early as possible, as it is poorly absorbed into the organism and should be provided in a pure form. The author also explains that tryptophan is not a viable precursor for psilocybin production due to end product inhibition in the biosynthetic process of P. cubensis. The author concludes that the main reason tryptamine works well, while tryptophan and DMT do not, is due to water solubility, as tryptamine is soluble in water and can be absorbed into the fungus during its normal biosynthetic process.

Believe me gray1, I have read the thread. I guess my mistake was believing Gartz, Shulgin, or YOU when you said:

' i would reccomend extracting the dmt and including it/exposing it to the fungi organism as early as possible. in order to be effective in increasing the psilocybin content (which we have shown numerous times as theoretically possible based on the simple cellular metabolic conversions) it must be bioavailable to the organism. i recently read a pare concerning incorporation of substances into the fungi based on radiolabelling, and it appears as if dmt is poorly absorbed/transported into the organism. therefore, make it as easy as possible by providing it in a pure form and exposing the organism to it as long as possible.

i also mention this in the doubters of the dmt method post
"tryptamine, whcih is readily formed from tryptophan by P. cubensis, serves as a better precursor of psilocybin than tryptophan. N-methyltryptamine is a still better progenitor of psilocybin, but n, n dimethyltryptamine is rather poorly incorporated as judged from the dilution factors, however, if the poor absorbtion of this compound by the fungus, less than 5%, is taken into account, the high dilution factor does not make it an unlikely intermediate." '

8^)

In reference to tryptophan, feel free to search the shroomery for the numerous posts about people trying to use tryptophan with NO PERCEIVEABLE increase in potency (Ryche Hawk for one). I will assume that perceivable is what counts here. I also understand that you can't trust everything posted on the shroomery, but judging the direction that people tend to lie around here, I would assume that if they were going to lie about this, they would claim it did work, not the other way around. And trust me, certain people would thrust themselves forward to write up a tek and claim credit for their discovery (sorry, that just amuses me to no end). I'm sorry, but I won't admit to having tried it, so I have no first hand experience with it. If you trust that I am not going to lie about it, I will say that I have spoken to a two organic chemists, and one biochemist who HAVE tried this (where it is legal to do so) and they had no luck. So again, if you trust me... Tryptophan (under normal conditions) will not boost psilo levels.

Ok, now I will hold your hand now and explain the "work" I was talking about. 8^)

I have theorized that there are genetic limitations in the biosynthetic process of P. cubensis that will try to hinder the production of excess psilo alkaloids. Do some reading on allosteric enzymes... The problem is end product inhibition. Basically when the amount of psilocin gets high, it starts binding to the enzyme converting tryptophan into tryptamine and prevents that step from occuring any further. The good news (and this explains the known behavior of adding precursors to P. cubensis) is that this usually only happens at the first step of the biosynthetic process. This partially explains why tryptamine results in massive psilocin, but tryptophan does not. Getting psilocybin, would probably require that work I was talking about. My theory WAS that you would have to supply not only enough nutrition for regular metabolic processes, but also EXTRA nutrition (maybe nutrition is not the best word here) to supply the building blocks of the excess: tryptamine, N-methyltryptamine, n, n dimethyltryptamine,psilocin, and psilocybin. We can be reasonable sure this is true for the psilocybin step. It probably doesn't have enough phosphate to create the psilocybin from the psilocin. My suggestion that starting with DMT would be easier meant that you would only have to supply this kind of extra 'nutrition' for the last two steps of the biosynthetic process since simply adding tryptophan isn't going to do it. So I was saying it would be more work to get tryptophan to work than to extract DMT. Looking at it now, we can see I was only partially right.. No matter what you do, you probably can't use tryptophan, but it still would require "something" extra to get tryptamine, or n-methyltryptamine to produce excess psilocybin and still be healthy enough to fruit normally.
That is all I was trying to say, and it really is a while lot of arguing over a petty issue. I think we have a lot more to offer each other working together on this. And even arguing we are comming up with some amazing ideas! The subject seems to have taken over the advanced forum !!!

Back to work:
Tryptamine results in few, gnarled, unhealthy looking mushrooms. Again probably not enough nutrition to carry out this much alkaloid production and maintain healthy growth, sporulation, and fruiting ability.

Now here is the BIG DEAL!!! I think I realized WHY tryptamine works so well and why dmt works only slightly

WATER SOLUBILITY!!!!

You might not be able to get the mushroom to take up what is in the substrate, but if it is in the water that that composes 92% of the fungus, then it will appear to be part of the mushrooms normal biosynthetic process, as it will be absorbed into the fungus..

Tryptamine is soluble in water, trytophan isn't, DMT is slightly, n-methyltryptamine is slightly...

What do you think?

OPI



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Re: MIMOSA BARK IN MAiLBOX!!!!! [Re: Opi]
    #336266 -

Opi great post I am no chemist but I would think that water solubility would be a important factor in the success of the experiment. I think that is why in the tryptamine cubinses tek they used tryptamine HCL instead of just plain tryptaime because the tryptamine HCL is water soluble and tryptamine alone is not.

"I have theorized that there are genetic limitations in the biosynthetic process of P. cubensis that will try to hinder the production of excess psilo alkaloids. Do some reading on allosteric enzymes... The problem is end product inhibition. Basically when the amount of psilocin gets high, it starts binding to the enzyme converting tryptophan into tryptamine and prevents that step from occuring any further. " Opi

I agree with you 100% here. I was reading some books on secondary metabolies and many of the pathways are limited by the end product. This allows the organism to spend energy one other things.

I posted something not to long ago about secondary metabolite overproduction take a look at it. Instead of trying to add some of these chemicals that are very hard to get and some may not even work we can have the fungi overproduce them for us. Industry does this all of the time. Trace metals play a very important role in many of these processes also industry limits certain nutrients to cause overproduction. Read that post it goes into more detail.





--------------------
_____________________________________________________________________________________
"The greatest enemy of knowledge is not ignorance, it is the illusion of knowledge." -Stephen Hawking

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Re: MIMOSA BARK IN MAiLBOX!!!!! [Re: Opi]
    #336562 -

very good point opi......

Keep Trippin' and Blazin'

-=SporeWorks=-


--------------------
Everything posted above was out of fun, none should be taken serious. I am currently under the influence so take that into consideration.

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Re: MIMOSA BARK IN MAiLBOX!!!!! [Re: Opi]
    #336910 -

opi- while i still don't aggree with your personal attack on me and your constant disregard for my maturity and intelligence (let me hold your hand... is that bullshit really necessary in the dialogue between two intelligent people?) i do aggree with (and have always comprehended) what you are saying scientifically, i still think that the extra work involved in trying a tryp experiment is minimal and more associated with creating a correct nutritional environment (including proper phosphate levels as that seems to be a concern for you) for the fungi, which is probably already created by a brf type substrate ---however, at this point i'm ready to chalk it up as a miscommunication of ideas and a difference of opioion rather than incorrect information by either of us, and as you suggested, we have more positive potential theorizing and experiemnting together than we do bickering about petty issues. i aggree with the solubility of DMT and the fact that this may play a role in it's poor incorporation into the organism, and was thinking of this when i suggested/posted the method for potency comparrison a few days ago. by liquid cultivating/allowing colonization, DMT may be more readily assimilated into the organism. -i also had an interesting thought earlier this morning when i remembered that for certain natural alkaloid biosynthesis in other organisms that certain stresses actually induces biosynthetic pathways (such as for phalaris and the prodcution of DMT) and should be part of the cultivation process. is it possible that this is also true for the biosynthesis of psilocybin/psilocin in fungi and that attempting to create and optimal environment for growth is actually counterproductive? it seems that tryp or another intermediate would still be necessary for the process, but as has been suggested, tryp probably already exists in sufficient amounts in typical growth substrates such as brf... any thoughts on this?


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Re: MIMOSA BARK IN MAiLBOX!!!!! [Re: gray1]
    #337046 -

Please, let me COMPLETELY appologize for insulting you. I honestly was trying to be funny (hence the smile faces) about the fact that I said I wouldn't explain it again, and felt drawn into doing it anyway. It wasn't a judgement on your intellegence or maturity. I just found it funny that there was a miscommunication (or inability on my part to communicate clearly) and I still felt a need to explain and re-explain. That was the joke. That was why it was funny to "pretend" it had to do with your intellegence. Regarless of whether I was trying to be funny or not, you were insulted and I am sorry.

"(including proper phosphate levels as that seems to be a concern for you)

Ha ha ha, I will never give up my quest for boosted psilocybin...

I don't think that BRF alone is enough, otherwise you would see the full genetic possibility of the species, huge fruit, max alkaloid content, etc. If we are going to elevate the concentration of one part of BRF we need supplement everything else the fungus is going to need to deal with that excess.

There are a couple good threads going on this subject right now, one of them dealing with adding zinc to the substrate. This sounds very promising. Bringing all of these ideas together might be a good idea. Maybe we should come up with a plan to test different ideas as well as combinations of ideas, to see what works. We ask for competent cultivators to act as virtual lab assistance and report their findings... Its rather subjective, but it might get us somewhere faster..

Opi


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Mushrooms, Mycology and Psychedelics > Advanced Mycology
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