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Invisiblemicro
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Re: Tryptamines in substrate with DPT? [Re: Coincidentiaoppositorum]
    #22255681 - 09/18/15 10:51 AM (8 years, 4 months ago)

Quote:

Coincidentiaoppositorum said:
4-HO-DMT is psilocin...

This is why this is all confusing to me, it seems like several different things have been jumbled together in a way to make it as unclear as possible, I'm interested in the topic here, but I'm having trouble trying to decipher your intent.....meaning I generally know the processes and enzymes here, but its hard for me to tell what exactly your trying to say.

There's a grid for the biosynthetic pathway of psilocybin because after tryptophan decarboxylation its unclear what pathways the fungi are using, there are a few diffferent possibilities...

It sounded to me like you were asking if by inhibiting hydroxylation and  Phosphorylation of the dimethyltryptamine if you could yeild it in the fruiting bodies...

But I got somewhat lost...

-E. Borodin




So, if the 4-hydroxylation were inhibited it might make DMT...

Or, it might skip over it entirely as it isn't "recognized" as a ligand any more.

Phosphorylation should be easy, just bump up the potency.

It has already been demonstrated this is the case.


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OfflineCoincidentiaoppositorum
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Re: Tryptamines in substrate with DPT? [Re: micro]
    #22255750 - 09/18/15 11:10 AM (8 years, 4 months ago)

A ligand has to do with  binding and binding affinity, it can be binding to other compounds or there can be binding to receptor sites....

.... which is why I'm having trouble following whats going on here, at times I think the biosynthetic pathway for psilocybin is being discussed, at other times its human metabolism of the compound being discussed, and at other times its chemical analysis or growing of the fungi being talked about ...sorry, I got lost at some point.

Sorry, you guys will have to let me figure out what your saying before I can participate productivly...




-E. Borodin


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Invisiblemicro
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Re: Tryptamines in substrate with DPT? [Re: Coincidentiaoppositorum]
    #22255776 - 09/18/15 11:16 AM (8 years, 4 months ago)

The enzyme has at least *some* specificity, right?

I don't know what to better call it.

My point was removing the hydroxyl group could cause the enzymes upstream to lose specificity.

If you can think of a better term I'm all ears.


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OfflineCoincidentiaoppositorum
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Re: Tryptamines in substrate with DPT? [Re: micro]
    #22256251 - 09/18/15 01:32 PM (8 years, 4 months ago)

Your going over my head, I'm trying to keep up though. I'm still learning all this as well.

-E. Borodin


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InvisibleToadstool5
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Re: Tryptamines in substrate with DPT? [Re: Coincidentiaoppositorum]
    #22258111 - 09/18/15 08:57 PM (8 years, 4 months ago)

Quote:

So, if the 4-hydroxylation were inhibited it might make DMT...

Or, it might skip over it entirely as it isn't "recognized" as a ligand any more.




Exactly it may create tryptamine, NMT, DMT, or perhaps some novel compounds not yet created.

We are talking about limiting the mushroom's metabolism while growing to yield different tryptamines.

Tryptophan is created in the mushrooms through the shikimate pathway, then it is decarboxylized to tryptamine. At this point it is either methylated by enzymes or oxidized by enzymes to yield the hydroxylized version of the tryptamines.

The proteins associated with the enzymes for the oxidation of the tryptamines and the enzymes themselves are not yet known.

I think they might be similar to other known tryptophan monooxygenase enzymes associated with cytochrome P450 proteins.

If we can inhibit the enzymes chemically without effecting or removing the proteins, we can alter the tryptamines produced by the mushrooms.


--------------------
If you do not know where the mushroom products you are consuming are grown, think twice before eating them. :badshroom:
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OfflineNear Dylan
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Re: Tryptamines in substrate with DPT? [Re: Toadstool5]
    #22327228 - 10/03/15 08:36 AM (8 years, 3 months ago)

Any progress on this experiment yet?


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InvisibleToadstool5
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Re: Tryptamines in substrate with DPT? [Re: Near Dylan]
    #22328783 - 10/03/15 03:04 PM (8 years, 3 months ago)

Nope, i am so poor i don't have the resources to start that type of project. Here and there I will try what i can using the methods to inhibit 5-hydroxy to see if they also have an effect on the 4-hydroxylation.

However, I think it will take more effort by the DIY bio movement to identify the gene responsible before we can hope to manipulate or shut it down.

I'm not educated in the ways of sequencing, transfers, isolation, and all that cool genetic modification stuff either so even if i had the lab and resources I would be going off of secondary research and hail-mary's. :lol:


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If you do not know where the mushroom products you are consuming are grown, think twice before eating them. :badshroom:
- Paul Stamets

AMU Teks :mushroom2: Stro's Write Ups


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Invisiblemicro
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Re: Tryptamines in substrate with DPT? [Re: Toadstool5]
    #22329261 - 10/03/15 04:34 PM (8 years, 3 months ago)

I read a journal article on this some time after the thread seemed to have died.

DPT *does* in fact work, and gets 4-hydroxylated.

N,N-DMT does not work very well; it causes inhibition of the metabolic pathway at some point.

Funny thing is, there is a similar thread from 2004 and I had posted in it; I just forgot :lol:

If you want to inhibit gene expression though, you would need more sophisticated resources.

I think the easiest way these days is with RNAi.


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InvisibleToadstool5
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Re: Tryptamines in substrate with DPT? [Re: micro]
    #22329732 - 10/03/15 06:43 PM (8 years, 3 months ago)

Quote:

I think the easiest way these days is with RNAi.




So true :shrug:

I have a feeling a genetics nerd will figure it out eventually and in the meantime it is a waiting game. There isn't enough focus on fungi, you know what I'm talking about micro. Even with something as common as mating types we lack serious research.


--------------------
If you do not know where the mushroom products you are consuming are grown, think twice before eating them. :badshroom:
- Paul Stamets

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Invisiblemicro
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Re: Tryptamines in substrate with DPT? [Re: Toadstool5]
    #22329844 - 10/03/15 07:14 PM (8 years, 3 months ago)

Yeah. I'm surprised, too.

You would think with all the money involved in commercial production someone would.

Especially since the mating-type loci control fruitbody production.

I'm hoping once I move I can try some stuff.

I haven't scored a whole lot of popularity with the neighbors playing mad scientist here :lol:

The people upstairs actually called the cops and told them I was insane and going to blow everything up.

Of course, the cops just thought she was a fruit loop which is p much the case anyway.


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InvisibleToadstool5
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Re: Tryptamines in substrate with DPT? [Re: micro]
    #22330731 - 10/03/15 10:56 PM (8 years, 3 months ago)

Quote:

The people upstairs actually called the cops and told them I was insane and going to blow everything up.




I know how that goes all too well hahaha

:guiltyascharged:


--------------------
If you do not know where the mushroom products you are consuming are grown, think twice before eating them. :badshroom:
- Paul Stamets

AMU Teks :mushroom2: Stro's Write Ups


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OfflineCraterBait
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Re: Tryptamines in substrate with DPT? [Re: Everything]
    #22352917 - 10/09/15 01:00 AM (8 years, 3 months ago)

Quote:

Everything said:
Basically, i am going to try this. or if a friend wants to i will give him the DPT to add to substrate.

Not even shulgin has info on dosage of 4-ho-DPT but i have a pretty good guess its around 25mg for a moderate to strong dose :wink:

This is a project in the distant future, nothing of now. but will report back if i have results. there is seriously no info on 4-HO-DPT because shulgin says it is so hard to make but i think he meant synthetically not through the mushroom, at the time of pihkal this method was not known.




I know in tikhal, page 247 starts a chapter 'dmt is everywhere' shulgin illustrates the properties of mycellia being able to 4-hydroxylate tryptamines with little bias and issues a few examples. It probably was indeed already known to shulgin.

My question to OP is, to help me understand his experiment: With so little information on 4-HO-DPT how do you plan on confirming its presence in the fungus? Furthermore; how do you intend on isolating it from other substances that may interfere with a bioassay.


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