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fastfred
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Registered: 05/17/04
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Discussion of Psilocybin pathway...
#5856702 - 07/14/06 01:03 AM (17 years, 6 months ago) |
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Quote:
Hey FF, just had a quick one for ya about your pathway diagram :
I'm not the most versed in chemistry, but I do understand most of the general concepts; with that I was wondering what process took place between each entity (ie psilocybin from psilocin from etc etc) I do know of how the body breaks down the psilocin, but how about all the other transitors? (I could be totally looking at this askew as well and not understanding it haha)
Where would Norbaeocystin ( C10 H13 N2 04 P ) fall into that flow?
Ugh stoner confusion, thanks for humouring me.
Tryptophan -> tryptamine -> N-methyltryptamine -> N,N-dimethyltryptamine -> Psilocin -> Psilocybin
It's decarboxylation, N-methylation, N-methylation, 4-hydroxylation, and O-phosphorylation.
Tryptophan decarboxylase is the only known and named enzyme. The others would be called N-methylase, 4-hydroxylase, and O-phosphorylase. That is assuming that they're site specific, which they probably are.
They might also be molecule specific which would cause the molecule to be named first followed by the reaction + ase. Such as "N-methyltryptamine methylase"
Good question about norbaeocystin. I kind of just threw baeocystin into the diagram to see where it would fit in. It's actually in the wrong place. Since it's already phosphorylated it probably won't be N-methylated, and it certainly won't be hydroxylated to psilocin. The arrow could go straight to psilocybin, but I don't think that's the case.
As far as norbaeocystin... If you add 4-hydroxytryptophan to the fungal culture you will get norbaeocystin and/or baeocystin.
So here's what I think happens... if tryptophan or tryptamine get 4-hydroxylated they then get phosphorylated to norbaeocystin and/or baeocystin. I'm not sure if they can then be N-methylated to psilocybin, but I doubt it. ------------------------------------------
I just thought I would post this since it seems like an interesting topic.
I have a couple scanned papers that I would like to post on this topic. Does anyone have Adobe Acrobat who would convert some .jpgs into .pdfs for me?
I'll attach the file to this post, if anyone can convert them for me it would be much appreciated.
-FF
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fastfred
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Re: Discussion of Psilocybin pathway... [Re: fastfred]
#5856708 - 07/14/06 01:07 AM (17 years, 6 months ago) |
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Here is the diagram that is referenced above...

-FF
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Feelers
Anti-Myth-Rhythm-Rock-Shocker


Registered: 06/18/02
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Re: Discussion of Psilocybin pathway... [Re: fastfred]
#5864453 - 07/16/06 06:17 AM (17 years, 6 months ago) |
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Cheers for the info FF. 
Has anyone ever tried up-regulation of any of the precursor genes? I don't spose there is much info about feedback mechanisms in the system?
Edited by Feelers (07/16/06 06:24 AM)
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fastfred
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Re: Discussion of Psilocybin pathway... [Re: Feelers]
#5865226 - 07/16/06 11:56 AM (17 years, 6 months ago) |
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Tryptophan is highly regulated. I don't think the other steps are well known enough to speculate much on them.
But, since addition of tryptamine increases psilocin, then the rest of the steps probably aren't regulated much if any.
Scientists have engineered tobacco to overproduce TDC (tryptophan decarboxylase) using agrobacterium. Agrobacterium can also be used on fungi.
If someone could just get their hands on the transformed agrobacterium that they used, it should be a simple matter to transform fungi. I'm not really sure how you would screen for functional inserts though... You might be able to measure growth on tryptophan supplemented medium, since increased TDC might cause a tryptophan shortage.
-FF
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mycogirl
goddamn



Registered: 07/03/05
Posts: 1,135
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Re: Discussion of Psilocybin pathway... [Re: fastfred]
#5873946 - 07/18/06 03:27 PM (17 years, 6 months ago) |
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The agrobacterium has a plasmid. It should be fairly accessible. Usually an antibiotic resistance gene is linked to the desired trait gene. This way you can screen with antibiotics for bacteria with your DNA.
As far as screening the transformed individuals, I have no idea what fungal people use. Maybe there is an anti fungal resistance gene inserted somewhere. This way they could grow in media supplemented with the anti fungal, which would kill untransformed individuals. (?)
You should be able to find some journal that has published their results.
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jellyfish

Registered: 10/02/05
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Re: Discussion of Psilocybin pathway... [Re: mycogirl]
#5878182 - 07/19/06 03:16 PM (17 years, 6 months ago) |
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If only there was a way to prevent the dmt from hydroxylation
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EFresh
Team Skanc

Registered: 07/21/06
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Re: Discussion of Psilocybin pathway... [Re: jellyfish]
#5892713 - 07/23/06 10:51 PM (17 years, 6 months ago) |
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would it be possible to use the agrobacterium to turn tryptophan into tryptamine which could then be added to the substrate?
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fastfred
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Re: Discussion of Psilocybin pathway... [Re: EFresh]
#5892745 - 07/23/06 11:09 PM (17 years, 6 months ago) |
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The TDC gene isn't expressed when it's in the agrobacterium plasmid. So no.
-FF
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Acinaxuz
In SomnisVeritas.

Registered: 06/20/06
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Re: Discussion of Psilocybin pathway... [Re: fastfred]
#5893973 - 07/24/06 10:37 AM (17 years, 6 months ago) |
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Quote:
You might be able to measure growth on tryptophan supplemented medium, since increased TDC might cause a tryptophan shortage.
That doesn't make sense to me exactly...
If TDC, as we had discussed before, were increased, the theory would be that with a good supply of tryptophan the decarboxylase would convert as it came because it was unregulated. Have you found more information on it that would show more insight? I'm not quite finished with the .zip yet, relation papers to go and what not.
Btw--thanks a ton for posting the papers, great reads!~
I've had to read back over most everything I had researched before, haven't looked at it for close to a week--feeling a bit behind.
-------------------- :~:~:~:~{ * }~:~:~:~:{ * }:~:~:~:~{ * }~:~:~:~: All posts are made with only the intent to entertain myself and should ONLY be read with the understanding that they are FICTICIOUS. I do not warrant information I provide for use in illegal activity of any kind nor do I condone it for any reason. Furthermore, I am not, I have never, nor will I in the future, take ANY part in illegal activites.
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cloudtop
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Registered: 08/16/04
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Re: Discussion of Psilocybin pathway... [Re: Acinaxuz]
#5904753 - 07/27/06 09:41 AM (17 years, 6 months ago) |
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BTW, unless I'm missing something in reviewing Wikipedia and other texts for background to this topic, the final conversion in the pathway takes place via a phosphatase rather than a phosphorylase. Correct?
http://en.wikipedia.org/wiki/Phosphorylation
-------------------- peacefromabovecloudtop
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fastfred
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Re: Discussion of Psilocybin pathway... [Re: cloudtop]
#5904787 - 07/27/06 09:56 AM (17 years, 6 months ago) |
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No. A phosphatase removes phosphate groups. From wikipedia...
"Phosphorylases are enzymes that catalyze the addition of a phosphate group from an inorganic phosphate (phosphate+hydrogen) to an acceptor."
"Do not confuse this enzyme with a phosphatase or a kinase! A phosphatase removes a phosphate group from a donor, while a kinase transfers a phosphate group from a donor (usually ATP) to an acceptor."
"The phosphorylases are named by prepending the name of the substrate, e.g. glycogen phosphorylase, starch phosphorylase, maltodextrin phosphorylase."
"All known phosphorylases share catalytic and structural properties."
So "psilocin phosphorylase" would be the name of the enzyme if it is psilocin specific. Since nothing is really known about the enzyme at this point it only gets the generic name "phosphorylase" IMO, but either one would be correct since either way you would be making an assumption about the nature of it's activity.
Good question though. It could also be a kinase, but there's no sense in getting all fancy with the descriptors until more is known about the biosynthetic pathway.
-FF
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cloudtop
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Re: Discussion of Psilocybin pathway... [Re: cloudtop]
#5904878 - 07/27/06 10:30 AM (17 years, 6 months ago) |
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Interesting. For some reason I had meant to type kinase and ended up w/ phosphatase after rereading the information a couple of times. At any rate, still good to clarify.
I'll probably fade into the background again as I admit my retention of chemistry knowledge seems to have been less than thrilling. I do have to wonder about the potential mechanism of bruising as it relates to the phosphorylation of Psilocin to Psilocybin, though, given anecdotal evidence that psilocybin-dominant mushrooms are less prone to bruising. It seems as though the byproducts of the phosphorylation process are likely culprits.
Here's hoping somebody figures this out.
-------------------- peacefromabovecloudtop
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fastfred
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Re: Discussion of Psilocybin pathway... [Re: cloudtop]
#5905171 - 07/27/06 12:26 PM (17 years, 6 months ago) |
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My theory on bruising is that cellular damage releases a witches brew of enzymes that cause enzymatic dephosphorylation of psilocybin to psilocin. The psilocin is then easily oxidized to the bluish product that I have dubbed "psilocybeen" in my diagram.
P. cubensis is known to have enzymes that will cause enzymatic dephosphorylation of psilocybin in warm aqueous solutions. Psilocin's oxidation product is also known to be bluish-green. The combination of those two facts seem to create a reasonable theory of bluing.
However, it could also be that there is enough psilocin present to cause the bluing reaction. An even more unlikely theory is that the bluing reaction is unrelated to the oxidation of psilocin.
-FF
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monstermitch
Growing in Bags Doesn't Work


Registered: 02/10/06
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Re: Discussion of Psilocybin pathway... [Re: fastfred]
#5905983 - 07/27/06 04:32 PM (17 years, 6 months ago) |
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rubbed with a touch of isopropyl.
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fastfred
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Re: Discussion of Psilocybin pathway... [Re: monstermitch]
#5906022 - 07/27/06 04:45 PM (17 years, 6 months ago) |
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Which one?
Haven't had any fruits on hand for years otherwise I'd be very interested in tests like that. Back in the day I did do some interesting ascorbic acid experiments. It will reverse the bluing reaction, but I never did figure out if it actually converted psilocybeen back into psilocin.
-FF
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monstermitch
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Re: Discussion of Psilocybin pathway... [Re: fastfred]
#5906048 - 07/27/06 04:52 PM (17 years, 6 months ago) |
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they are both the same fruit.
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monstermitch
Growing in Bags Doesn't Work


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Re: Discussion of Psilocybin pathway... [Re: monstermitch]
#5906054 - 07/27/06 04:53 PM (17 years, 6 months ago) |
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so if I do this again to another fruit,
and then rub with ascorbic acid,
it shoud return to normal flesh tone?
killer. I'm so all over this.
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fastfred
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Re: Discussion of Psilocybin pathway... [Re: monstermitch]
#5906158 - 07/27/06 05:36 PM (17 years, 6 months ago) |
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Who knows? If you put a normally blued mushie into an ascorbic acid solution it will return it to normal color.
So I take it that the top pic was the one rubbed with IPA?
-FF
-------------------- 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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monstermitch
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Re: Discussion of Psilocybin pathway... [Re: fastfred]
#5906179 - 07/27/06 05:49 PM (17 years, 6 months ago) |
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they are both the same fruit.
it is just different perspectives of the same fruit.
the fruit in both pictures was rubbed with a tad of 80% isopropyl.
I'll make some ascorbic acid solution some time soon and try it.
--------------------
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fastfred
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Re: Discussion of Psilocybin pathway... [Re: monstermitch]
#5906280 - 07/27/06 06:28 PM (17 years, 6 months ago) |
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Quote:
monstermitch said: they are both the same fruit. it is just different perspectives of the same fruit. the fruit in both pictures was rubbed with a tad of 80% isopropyl. I'll make some ascorbic acid solution some time soon and try it.
If they are both the same fruit and they were both rubbed with 80% IPA, then why is one nearly black while in the other pic it looks normal?
I'm missing something here, please try to explain it to me.
-FF
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