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Feedback inhibition (or end product inhibition) is a mechanism for the inhibition of preformed enzymes that is seen primarily in the regulation of whole biosynthetic pathways, e.g. pathways involved in the synthesis of the amino acids. Such pathways usually involve many enzymatic steps, and the final (end) product is many steps removed from the starting substrate. By this mechanism, the final product is able to feed back to the first step in the pathway and to regulate its own biosynthesis.

In feedback inhibition, the end product of a biosynthetic pathway inhibits the activity of the first enzyme that is unique to the pathway, thus controlling production of the end product. The first enzyme in the pathway is an allosteric enzyme. Its allosteric site will bind to the end product (e.g. amino acid) of the pathway which alters its active site so that it cannot mediate the enzymatic reaction which initiates the pathway. Other enzymes in the pathway remain active, but they do not see their substrates. The pathway is shut down as long as adequate amounts of the end product are present. If the end product is used up or disappears, the inhibition is relieved, the enzyme regains its activity, and the organism can resume synthesis of the end product. Thus, if a E. coli bacterium swims out of a glucose minimal medium into milk or some other medium rich in growth factors, the bacterium can stop synthesizing any of the essential metabolites that are made available directly from the new environment.

One of the most intensely studied bacterial pathways is the pathway of tryptophan biosynthesis (Figure 3). The pathway of tryptophan biosynthesis is regulated by feed back inhibition. Tryptophan is the effector molecule for allosteric enzyme a. When the end product of the pathway (tryptophan) attaches to enzyme a, the enzyme is inactive and can no longer join glutamine and chorismic acid into anthranilate. If tryptophan is disjoined from the enzyme the pathway is resumed, and tryptophan synthesis will continue. Tryptophan biosynthesis is also regulated at a genetic level by the processes of enzyme repression (below) and attenuation.

Note: In the case of feedback inhibition (above), the signal molecule, tryptophan, is a negative effector of Enzyme a in the pathway of tryptophan biosynthesis, because when it binds to Enzyme a, it inactivates the enzyme. In enzyme repression (below) tryptophan is a signal molecule that acts as a positive effector of the trp repressor protein because when it binds to the repressor it activates the protein, so that it binds to the trp DNA.

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Phenotypic Effects of TDC Expression
T0 and T1 tobacco plants accumulating TDC in the chloroplast showed a striking phenotype. One or 2 weeks before the flower buds developed, small necrotic areas appeared on the surface of older leaves (Fig. 5A). With the onset of the opening of fully developed flower buds, the lesions increased in number and size in the lower and middle leaves, whereas younger leaves showed only small necrotic areas at the leaf edges. At the end of the flowering, most of the leaf surface was necrotic, and strong deformation was apparent in the lower and middle leaves (Fig. 5, B and C). These symptoms were observed only in plants expressing chloroplast-targeted TDC and not in plants expressing TDC in the cytosol or ER. In addition, within the group of plants expressing chloroplast-targeted TDC, only those shown to accumulate high levels of TDC and tryptamine displayed symptoms, indicating a correlation between symptom severity and the level of enzyme and product.
T0 plants with necrotic leaves were fully fertile and produced seeds that were used to generate T1 plants that showed the same phenotype displayed by T0 plants and no significant reduction of fertility.

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(I found this on a .gov site, pub med central, no spaces--google it if interested.)

Over-running the enzyme and it resulting in necrotic spots makes sense in literal ways, but the chemical reaction is beyond me. I'm thinking that if plant is overloaded with TDC, it then begins attacking itself due to limited supply of proteins (or needed addition to complete synthesis).

The same indeed could be true in mushrooms, applying the same logic it could eat "itself" to produce psilocin. I wonder if there's anything to this theory when it comes to hollow stems? There is a strain that is known for hollow stems--maybe it's because there is an overage of TDC. Has anyone attempted growing that strain (can't think of it at the moment) on heightened levels of tryptophan? (Oyster Shells, Seaweed, B vit additions etc, how did it perform on poo?) A large supply in theory would feed the enzymes. Could it actually be an over supply of enzyme we're still stuck with the same issue of providing abundant tryptamine to feed the need. Damn, I can't win for losin.

Ok, so in theory, a source of tryptamine AND a source of TDC could be incorporated for sturdier results.

Personally, I say stick with one thing at a time... back to the drawing board.

(ps fred--thanks for the replies, it really helps the thought process, keeps me grounded )

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Because endogenous TDC activity was not detected in the present study or in previous studies in tobacco, the biosynthesis of tryptamine in infiltrated leaves transiently expressing targeted TDC was used as a direct evidence of in vivo enzyme function. Tryptamine levels in infiltrated leaves were measured using a fluorometric assay (Sangwan et al., 1998), and the results (fluorescence intensity per gram of fresh weight leaf material) are shown in Figure 2. Leaves transiently expressing chloroplast-targeted TDC showed a fluorescence intensity of tryptamine approximately 1.5-, 5-, and 72-fold higher than those expressing TDC in the cytosol, in the ER, or the background levels of control leaves infiltrated with nonrecombinant Agrobacteria, respectively (Fig. 2). Nevertheless, tryptamine fluorescence in leaves transiently expressing cytosolic or ER-targeted TDC was approximately 48.5- or 13.5-fold higher than the background signal of control leaves, respectively (Fig. 2).

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What's interesting is the difference in chloroplast and non-chloroplast targeted TDC. They must have screwed around with transforming chloroplast DNA?

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Note: In the case of feedback inhibition (above), the signal molecule, tryptophan, is a negative effector of Enzyme a in the pathway of tryptophan biosynthesis, because when it binds to Enzyme a, it inactivates the enzyme. In enzyme repression (below) tryptophan is a signal molecule that acts as a positive effector of the trp repressor protein because when it binds to the repressor it activates the protein, so that it binds to the trp DNA.

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>>edit<< Did a lot of re-reading--first link, next post--poses better clarification! >>edit<<

I'm so glad that this drive for information is up and running! I can't wait to see some real results from all of this hard to come by information! I am a firm believer in the medicinal value of mushrooms, and seemingly throughout history almost every culture welcomed them, except for us... think of the benefits this could hold for medicine sake.

Ok, yea I know, I'm a little over dramatic, but, at least I'm an extremely excited loser with a purpose!  lol

btw, thanks for the shrooms I always enjoy good vibes and I'll be happy to throw them back your way!

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Over-running the enzyme and it resulting in necrotic spots makes sense in literal ways, but the chemical reaction is beyond me. I'm thinking that if plant is overloaded with TDC, it then begins attacking itself due to limited supply of proteins (or needed addition to complete synthesis).

The same indeed could be true in mushrooms, applying the same logic it could eat "itself" to produce psilocin. I wonder if there's anything to this theory when it comes to hollow stems? There is a strain that is known for hollow stems--maybe it's because there is an overage of TDC. Has anyone attempted growing that strain (can't think of it at the moment) on heightened levels of tryptophan? (Oyster Shells, Seaweed, B vit additions etc, how did it perform on poo?) A large supply in theory would feed the enzymes. Could it actually be an over supply of enzyme we're still stuck with the same issue of providing abundant tryptamine to feed the need. Damn, I can't win for losin.

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Coffee is not added for potency just growth speed, I have done my own tests so have many others and it appears to work, I have seen some deformities but few, there is no marked effect on potency.

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Notice the necroptic spots on the caps of the mushie, note, not nipples... deformations, breaking down of tissue...

It seems the coffee grounds have sped growth significantly, but the effects remain, as other testing done with coffee is not resulting in these spots! Look at all of the aborts...

Since the growth was sped up by the stimulant to balance growth there dwarving occured slightly less, HOWEVER, the accumulation of tryptamine still made for necroptic spotting on the caps and abundant aborts. My conclusion would be that the mushroom is reacting to the same enzyme process as the tobacco plants.

I still say it's a reaction because each individual environment is different... it poses different hazards for different biochemical make-ups, the presence of one enzyme may make the compound stable, and the lack of that enzyme in another environment could lead to breakdown and rebonding, leaving it ineffective to continue it's processes. This of course would result in "dying" portions of the mushroom...

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I thought you would like to see these pics of some z-strain I grew with coffee grounds added, Nice smooth caps

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those are very pretty, and smooth!  Those weren't grown with the acacia though, right?

What I'm pointing out, is that the deformity isn't coming from the coffee... it's coming from the TDC that's also found in acacia.

Summary:  TDC changes Tryptophan into tryptomine then tryptomine is changed to psilocin. The previous theory is that if you add tryptomine or tryptophan into substrate, you would receive higher potency--which is true to an extent. The mushroom regulates how much chemical it makes based on chemical levels--just like any other living organism, plant or animal. So by increasing tryptophan or tryptomine, you're not doing anything for the mushroom because the mushroom can and does make this on its own. If you feed the mushroom what it normally makes, it saves energy and utilizes what's in the substrate--thus, it does not produce "extra" for higher potency production... This opens up a whole other can of worms...

Everything grows in cycles, the mushroom's cycles are a series of created chemical compounds made specifically for a "job". In mushrooms tryptophan is produced to merge Chorismate and Glutamine (see post that starts with "I figured out why tryptophan won't work", above for further detail.) which in turn does another job, merges, and does another job. There's all kinds of really kick ass information in the links up there and if you're interested it's there for the clicking. Moving on...

If you increase the amount of TDC (Tryptophan Decarboxylase--enzyme which breaks down Tryptophan into tryptamine) you would in essence have continual increase in psilocin, because unlike tryptophan or tryptamine it isn't regulated. As the mushroom "eats" and absorbs, and the compounds inside of it chemically change, it only produces as much as the mushroom itself can regulate. Meaning, there is a maximum amount of tryptophan that the mushroom can keep in its system.

As far as we know at this point, there is NO limit to TDC which would create tryptomine/psilocin. As long as there is enzyme there (though from another plant source) it still, in the long run, suffices to make an over abundance of psilocin. This has been done in tobacco plants with a fair amount of success (increasing tryptamine, not psilocin)--though the benefits there are lost, when clearly the benefits here are plenty.

The issue comes in with the side effects of the over stocked tryptamine. Tobacco plants fed extracted TDC from another plant, it began getting "rot spots" from the heavily increased chemical.

My theory on this is just that it's from another source--certain enzymes, or inhibitors, or even effectors could alter the chemical enough to bind/merge with something else to inactivate it, which is somewhat supported by the links above, but is still in research status. 

Previously, the development path taken by the mushroom was not generally thought to contain similar tryptamines to serotonin, but as Dr. Albert Hofmann found, the chemical psilocin is only 1 hydroxyl molecule away from our very own serotonin.

Serotonin is used in the body to regulate our temperatures and sleep cycles and acts as a neurotransmitter. Being this the case, and that psilocin breaks down in our body, enzyme inhibitors (though I admit I have no idea as of yet what chemical it would be) in a sense shut everything down and then you experience the distorted imagry and "spiritual connection" most find in psilocin/psilocybin intake.

I believe that psilocin is the same in mushies, and seemingly, the idea also has FF's brain going, whom by the way, I appreciate dearly! Being the bundle of brains, experience, and chem. books that he is, he may even be able to shed more light on the biosynthetic pathway.

The title kind of throws this off a bit--and it seems that we're not getting too many people due to the topic being beaten to a pulp... but as far as I know, this is the only one of the kind.

If anyone has info--POST IT! No sense in wasting time looking for answers that we've already found to be absolute. Use research to debunk this thread... the more we know it's not, the better our chances of finding what is. Throw in ideas!... something!... it's getting lonely in here!

<3 -Zuxi