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OfflineInterested
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Nitrogen as key to potency
    #4634131 - 09/08/05 04:32 PM (11 years, 10 months ago)

People seem to generally believe that nitrogen content, in some form or another, is the key (to a point, at least) to high alkaloid content in psychoactive mushrooms. This is frequently cited as the explanation for the high potency reported in mushrooms grown on a manure substrate.

Can anyone provide a source (preferably a journal article, obviously) for this idea? "High nitrogen content" clearly isn't much to go on. In particular, it leaves one wonder in what form the nitrogen must be in order for the fungus to make use of it.

Any journal references for this nitrogen content business would be greatly appreciated.


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Offlineamyloid
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Re: Nitrogen as key to potency [Re: Interested]
    #4635909 - 09/09/05 12:09 AM (11 years, 10 months ago)

i only respond to this because no one else has yet....

ive never seen nitrogen being listed as any kind of factor in potency any scientific/otherwise text.

also, arnt most substrates abundant in nitrogen? i think most nitrogen is available through nitrogen compounds, im gonna take a wild guess and say something like ammonia cause i know thats in fertilizers.

the only thing i know about nitrogen's relation to alkaloids is that alkaloids have like two nitrogen molecules or something..referred to as nitrogenous. forgive me cause im by no means any kind of chemist.

here is a little reading material so you can read more in depth.

http://en.wikipedia.org/wiki/Alkaloid

http://en.wikipedia.org/wiki/nitrogen


--------------------
"A human being is part of a whole, called by us the Universe, a part limited in time and space. He experiences himself, his thoughts and feelings, as something separated from the rest--a kind of optical delusion of his consciousness. This delusion is a kind of prison for us, restricting us to our personal desires and to affection for a few persons nearest us. Our task must be to free ourselves from this prison by widening our circles of compassion to embrace all living creatures and the whole of nature in its beauty."
-Al Einstein


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Offlinebloodbob
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Re: Nitrogen as key to potency [Re: amyloid]
    #4636384 - 09/09/05 01:51 AM (11 years, 10 months ago)

Yeah basicly if you don't have a high enough amount of nitrogen your mushrooms can't produce alkoloids.


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Offlineblackout
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Re: Nitrogen as key to potency [Re: bloodbob]
    #4636937 - 09/09/05 06:27 AM (11 years, 10 months ago)

I will be doing some comparisons soon (not chem analysis, just peoples opinions and differences in blueing)

http://www.shroomery.org/forums/showflat.php?Cat=0&Number=4493850


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Edited by blackout (09/09/05 06:28 AM)


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Offlinescatmanrav
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Re: Nitrogen as key to potency [Re: blackout]
    #4637494 - 09/09/05 12:54 PM (11 years, 10 months ago)

Mushrooms change the nitrogen into tryptophan, or whatever that is..adding a high nitrogen source to poo boosts potency..bloodmeal, bonemeal, or my favorite, worm castings all work for alot of nitrogen. I have no proof, other then I know high nitrogen substrate produce mushrooms that make people whine at me that they are to potent..


--------------------
"life is like a drop of rain getting closer and closer to falling into a lake, and then when you hit the lake there is no more rain drop, only the lake."

Growing with bags, start to finish (including my new grain and substrate prep)
Anyone looking to start bulk tubs/mono tubs/shotgun hybrids? Good tubs to use..
How I do grain (old still good tips)
Turn your closet into a fruiting chamber
Casing layer colonization and overlay


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OfflineInterested
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Re: Nitrogen as key to potency [Re: scatmanrav]
    #4637562 - 09/09/05 01:14 PM (11 years, 10 months ago)

I understand that it's generally accepted that nitrogen leads to high alkaloids levels. My particular interest is in, aside from personal experience, the source of this knowledge -- journal articles or book references. Aside from simply confirming its truth, this would likely provide insight as to what form the nitrogen must be in in order to be bioavailable. This is what I said in my first post.

amyloid: Thanks for the links, but I think we're talking at different levels. I'm have a pretty reasonable background in biochemistry, and in fact wrote a portion of the alkaloid entry you link to. My problem is not misunderstanding what nitrogen or alkaloids are.

It's true that all alkaloids have at least one nitrogen atom in them, but this alone does not explain why high nitrogen levels lead to higher potency. A very large portion of fungal mass is in proteins, each of which contains thousands of nitrogen atoms. The nitrogen consumed in protein production easily dwarfs (by many order of magnitude) that needed for alkaloid production. Thus, if its growing, it has access to large quantities of nitrogen.

This is not to say that high nitrogen levels don't lead to increased alkaloid production -- I assume this is true, which I why I'm interested in the matter. There's a lot more to it, though, than the fungus simply needing the nitrogen to toss into the alkaloid molecules.

scatmanrav: The nitrogen actually comes in a fair bit earlier in the biosynthesis pathway than tryptophan. The indolic structure itself, upon which tryptophan is based, contains nitrogen. As I say, though, nitrogen is present in nearly every molecule in the fungus.

Again, to repeat myself, my interest is in identifying the original source for this idea -- a book, or journal article. It may simply be "common knowledge", but if this is the case it would be helpful to know this as well.


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Offlinescatmanrav
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Re: Nitrogen as key to potency [Re: Interested]
    #4637619 - 09/09/05 01:31 PM (11 years, 10 months ago)

The "original source" would probably be agar...talk to him for more info...but I havent seen him around much latly, sorry I dont know the "original source" but agar probably would, hes worked alot with creating compost containing everything shrooms need and figuring out what shrooms need. I have only taken as is, and accepted it, never found the actual original source..but as I said, agar is the one who has talked about it the most.


--------------------
"life is like a drop of rain getting closer and closer to falling into a lake, and then when you hit the lake there is no more rain drop, only the lake."

Growing with bags, start to finish (including my new grain and substrate prep)
Anyone looking to start bulk tubs/mono tubs/shotgun hybrids? Good tubs to use..
How I do grain (old still good tips)
Turn your closet into a fruiting chamber
Casing layer colonization and overlay


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InvisibleHermes_br
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Re: Nitrogen as key to potency [Re: Interested]
    #4637638 - 09/09/05 01:40 PM (11 years, 10 months ago)

Quote:

2. Low levels of ammonium succinate, which will give poor yelds of psilocybin






:mushroom2:


Edited by Baby_Hitler (09/11/05 03:28 PM)


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OfflineInterested
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Re: Nitrogen as key to potency [Re: Hermes_br]
    #4637665 - 09/09/05 01:51 PM (11 years, 10 months ago)

This (or more accurately, reference #10) is EXACTLY what I was looking for. I cannot thank you enough Hermes_br.

I should have known that someone quoting Colin McGinn in their sig would be able to help. :-)


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OfflineInterested
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Re: Nitrogen as key to potency [Re: Interested]
    #4637678 - 09/09/05 01:57 PM (11 years, 10 months ago)

On an extremely embarrassing note, I've read the article before.

http://www.shroomery.org/forums/showflat.php/Number/4081322

Fuck.

If anyone would like to read a PDF copy of the article (flipping through those GIFs was driving me crazy) I've stiched one together at: http://rapidshare.de/files/4907484/production-submerged.pdf.html


Edited by Interested (09/09/05 02:11 PM)


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Offlinestvip
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Re: Nitrogen as key to potency [Re: Interested]
    #4637877 - 09/09/05 03:00 PM (11 years, 10 months ago)

All good and well, except that the study results don't actually support the suggested conclusion ("... Low levels of ammonium succinate, which will give poor yelds of psilocybin" refers to reduction to a subcritical level - omission, whereas the referenced study explicitly notes "In contrast, levels of ammonium succinate which were one-half or twice that of the normal medium had no significant effect upo pH or psilocybin production")
Thanks a lot fo posting the article! Though I'm not really sure conclusions from studies of dried mycelium pellets from submerged cultures have strong enough implications for what one might find in the basidocarps of mature specimen (there's huge variation throuh days 5-11 alone, and I also have doubts about their extraction efficiency of intracellular meterial).


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OfflineInterested
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Re: Nitrogen as key to potency [Re: stvip]
    #4637948 - 09/09/05 03:12 PM (11 years, 10 months ago)

Extraction efficiency shouldn't matter terribly, though. As long as its consistently efficient, the results are valid.

I disagree about the results not showing (or, at least, suggesting) that conclusion. The levels of ammonium succinate present in their normal medium may well be substantially higher than the levels occurring in solid media. I see no reason to think that nutrient content in their normal media corresponds (or is meant to) to nutrient content in standard growing media. It's entirely possible that, for example, brown rice flower contains ammonium succinate in only subcritical levels.

I'd be curious to know why they chose ammonium succinate, and may look through their references to see if I can find out. Was it simply an arbitrary source of ammonium, or is ammonium succinate particularly bioavailable to the mycelium?


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OfflineInterested
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Re: Nitrogen as key to potency [Re: Interested]
    #4638730 - 09/09/05 06:22 PM (11 years, 10 months ago)

Journal article request! Can anyone come up with:

Leung, A.Y. and Paul, A.G.. "The Relationship of Carbon and Nitrogen Nutrition of Psilocybe baecocystis to the Production of Psilocybin and its Analogs," Lloydia, Vol. 32, No. 1(March, 1969), 66-71.

The Journal of Natural Products website doesn't have backissues this far, so I suspect they don't exist in electronic form. If anyone has access to a dead tree copy that they could scan, though, it would be very helpful.


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InvisiblePinback
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Re: Nitrogen as key to potency [Re: Interested]
    #4638795 - 09/09/05 06:38 PM (11 years, 10 months ago)



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InvisiblePsychoslut
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Re: Nitrogen as key to potency [Re: scatmanrav]
    #4638909 - 09/09/05 07:13 PM (11 years, 10 months ago)

The source for that information is The Mushroom Cultivator- Paul Stamets and J.S Chilton.


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[quote]KristiMidocean said:
Good now thats clear.WHO FUCKING CARES. If I am fat u all keep pointing it out like its suppose to be a secret.LIke u really have nothing better to do then make fat jokes. If o know its like I do I know yall can come up with NEW AND BETTER SHIT . This shit is old and boring . I left in the first place cause this shit got boring not because of the fat jokes . Fat jokes dont bother me but seriously its old[/quote]


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Offlinestvip
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Re: Nitrogen as key to potency [Re: Pinback]
    #4638955 - 09/09/05 07:25 PM (11 years, 10 months ago)

Quote:

Extraction efficiency shouldn't matter terribly, though. As long as its consistently efficient, the results are valid.




If there are significant different extraction efficiencies for intracellular and extracellular alkaloids, results will be misleading inasmuch as the ratio of the two varies.

Quote:

It's entirely possible that, for example, brown rice flower contains ammonium succinate in only subcritical levels.




Well, it shouldn't contain any ammonium succinate. But as a nitrogen source, note that the normal concentration of their nutrient media is 1 gram of AS and 9 grams of glycine per 1 liter medium, so at a ballpark estimate, BRF should have a sufficient nitrogen content. As the ommission of AS versus glycine shows, the type of nitrogen carrier is of utmost importance - removal of glycine increased psilocybin content, whereas removal of AS decreased it (despite the fact that glycine is the preponderant nitrogen source). If anything, the study implies nitrogen content beyond minimal is, in itself, is of no predictive value (but of course, the extreme case nitrogen starvation will inhibit all growth).


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OfflineInterested
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Re: Nitrogen as key to potency [Re: stvip]
    #4639193 - 09/09/05 08:23 PM (11 years, 10 months ago)

Pinback:
Thanks! Much appreciated.

stvip:
The article indicates that no extracelluar psilocybin was found. Or do I misunderstand you?

Thanks for the, uh, charitable interpretation of my second comment. Yes, I meant "contains a nitrogen source in only subcritical levels". I shouldn't be allowed to post before coffee.

Utter nitrogen starvation will, of course, prevent growth. I agree that it implies that nitrogen content alone cannot be used to predict alkaloid levels, but it's clear that ammonium succinate (in that study) content can -- at least, within a certain range of ammonium succinate levels. (Past which, as you say, it doesn't seem to do much of anything.)

Anyway, I think we agree, albeit with confusion caused by my pre-coffee idiocy.

The relevant question now would seem to be what the nitrogen must be bonded to in order for it to be used in alkaloid production. This is the second question I'd asked in my first post, and one I'd expected the article to answer.

Any thoughts? So far we've got:
Ammonium succinate: Yes
Glycine: No

Not exactly a huge dataset.

I'm going to start digging through this ( http://www.genome.jp/dbget-bin/show_pathway?MAP00380+1.13.11.11) nightmare to see if it provides any insight on the nitrogen source requirements. The other likely source for an answer would be to figure out what the authors of the study read that convinced them that ammonium succinate was suitable. Unless it's a standard ingredient in these sorts of things?


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OfflineInterested
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Re: Nitrogen as key to potency [Re: Interested]
    #4639236 - 09/09/05 08:37 PM (11 years, 10 months ago)

Having now read the study that Pinback provided, our dataset turns out to be substantially larger. This provides little insight on the general question of what sort of nitrogen source can be directed toward alkaloidal growth, however, and the authors of the study appear as uncertain as I am.

Damn.


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OfflinePinx
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Re: Nitrogen as key to potency [Re: Interested]
    #4643146 - 09/10/05 08:30 PM (11 years, 10 months ago)

There is something mentioned in "The Fungi" by Michael J Carlilie either 4th or 5th printing of the 1st edition i think page 128:

"""Carbon/Nitrogen Ratio

Proteins contain about 15% nitrogen. ... A balanced medium will hence contain about ten times as much carbon as nitrogen. Hence Carbon:Nitogen ratios of 10:1 or less will ensure a high protein content..."""

-)The term medium suggests that hes talking about a liquid substrate but i think this should also be true for solid substrates.
-)(An exception are Wood loving species since on wood the C/N Ratio is about 500:1.)

A high protein content ensures the producion of a high concentration of (aromatic) aminoacids wich are used for the production of proteine and peptides as primary and alkaloids etc as secondary metabolites.

possible nitrogen sources would be salts containing ammonia or nitrogen or both depending on the desired pH one would need to make a few calkulations but that is just basic chemistry..

Grains usually contain 10% Protein lets say 15 to get nice numbers...
15% Protein means 1% Nitrogen

And Grains contain betwenn 35 and 45 % Carbon(not Carbonhydrates..)
wich means the C/N Ratio is in the best case 35:1

While a C/N ratio of 50:1 is good for the production of organic acids, alkohol etc. it is pretty bad for the production of Proteins.

This leads me to the conclusion that there is an enormous potential of increasing the psylocibin production of mushrooms cultivatet on grain by adding an Nitrogen soure wich brings the C/N ratio to 10:1 the
problem is that one would need to add around 3g of ammonnitrate per 100g of grain wich could lead to a water problem.

Another idea im not able to try at the moment is the use of legumes(beans..) as substrate since their protein content is nearly twice as hish as the protein content of grains and im pretty sure, that the fungus should be able to utilize the carbon hydrates from the legumes...

i hope its at least readable and i didn t mess it up since it took me one and a half hour to get all this information together and write it down here...

greets

btw ammonia is a sutiable nitrogen source for 99.95% of the fungi as well as aminoacids and proteins are im not soo sure about nitrate but im pretty confident, that it should work..

and i dont think the nitrogen source has ANY influence on the building of alkaloids since the metabolistic cycles turn pretty much everything into what THEY need and high concentrations of tryptophan for instance will be turned into other amino acids rather fast..


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OfflineProsgeopax
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Re: Nitrogen as key to potency [Re: Pinx]
    #4643294 - 09/10/05 09:17 PM (11 years, 10 months ago)

Try adding ammonium phosphate or diammonium phosphate, they are used in beer making to kick start the yeast. They are good sources of easily utilized nitrogen as well as phosphorous. They are available at home brewing supply stores and are sometimes referred to generically as 'yeast nutrient' - but there are other 'yeast nutrients' which are basically dead yeast or a combination of dead yeast and one of the ammonium phosphates or even some b-vitamins. The ammonium phosphates are a white powder.


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You can accept, reject, or examine and test any new idea that comes to you. The wise man chooses the third way.
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Disclaimer: I reserve the right to change my opinions should I become aware of additional facts, the falsification of information or different perspectives. Articles written by others which I post may not necessarily reflect my opinions in part or in whole, my opinions may be in direct opposition, the topic may be one on which I have yet to formulate an opinion or have doubts about, an article may be posted solely with the intent to stimulate discussion or contemplation.


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Mushrooms, Mycology and Psychedelics >> Advanced Mycology

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