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DMT yeast/ecoli/anyone with genetics equipment? Options
 
John.Brown
#1 Posted : 11/10/2021 3:51:06 PM
i am curious if anyone on the forum has access to genetics equipment to possibly make a dmt producing yeast/e coli as it would only require the addition of 2 enzymes. INMT and AADC. yeast would be easier fir the average person to keep alive but e coli would definitely be easier to modify. i know there are kits to do this but i am so poor rn lol. ive seen some posts on here about it before but theyre almost 10 years old. it would be great for a bunch of people to pitch in to get the code written and inserted even and they just propagate and distribute the yeast. there are some services online that will even send you a yeast sample with the integrated plasmid if you can afford it.
 
Spiralout
#2 Posted : 11/11/2021 11:23:21 PM
I remember people talking about idea's surrounding this offf and on over the years. I can't remember the specifics though (because this is waay over my head) and can't remember any recent talk.

It's definitely interesting though. I'm sure someone will chime in with the latest news. Do you have links to more info about the kits you're talking about?
 
Dirty T
#3 Posted : 11/12/2021 3:12:31 AM
There is a guy on Shroomery that was working on psilocybin yeast a while back, I also know there is a company that has psilocybin yeast as well they are selling pure psilocybin for medical research. Yeast is by far the most efficient way to make huge amounts of pure psilocybin. If the same technology can be applied to produce DMT it would be amazing but scary if it falls into the wrong hands.
 
endlessness
Moderator
#4 Posted : 11/12/2021 2:27:16 PM
AFAIK this was not completed by any of the members previously discussing due to... well.. life getting in the way

But it's possible, and it gets easier each year with technology advancing...
 
benzyme
Moderator | Skills: Analytical equipment, Chemical master expertExtreme Chemical expert | Skills: Analytical equipment, Chemical master expertChemical expert | Skills: Analytical equipment, Chemical master expertSenior Member | Skills: Analytical equipment, Chemical master expert
#5 Posted : 11/12/2021 11:56:56 PM
we threw the idea around, and concluded that it's prohibitively expensive to source the mRNA/cDNA kits. The technology may have improved over the past decade, but the price hasn't; that's the caveat to dealing with oligopolies.

I could talk to my peers, if I remember; they'd certainly be able to initiate such a project, they're biohackers. We've talked about it before. They have genetic engineering equipment, as do I (including a couple thermalcyclers).
"Nothing is true, everything is permitted." ~ hassan i sabbah
"Experiments are the only means of attaining knowledge at our disposal. The rest is poetry, imagination." -Max Planck
 
Spiralout
#6 Posted : 11/14/2021 11:36:10 PM
@Dirty T : So what is the name of the company? If they provide psilocybin for research purposes they must really be on the up and up, given the tiny, restricted scope of research. I would guess they are producing the psilocybin they have for sale via the yeast?

What would the method be? Do you use some kind of CRISPR tech to engineer it?

 
Dirty T
#7 Posted : 11/16/2021 5:09:14 PM
I dont remember the exact name but they were in Cahoots with Compass Pathways and they consider their yeast a 'proprietary formula' which I'm sure is why they are selling the Psilocybin at godawful prices and I am equally sure there is no way to buy yeast from them unless you are one of the two companies allowed to have it as it would be the equivalent of live mycelium which contains psilocybin and is the reason that spores are legal but live culture is not unless you live in someplace like Sweden.
 
John.Brown
#8 Posted : 11/17/2021 3:46:37 PM
if some of you wonderful people actually have genetics equipment, it wouldnt be hard to get insert a plasmid into yeast, genscript is one of the services that will design a plasmid for you. it costs like 1k usd but if a bunch of people pitched in it wouldnt be impossible. it could be cheaper now idk, i havent messed with this stuff in a bit. it would be much cheaper to do the insertion yourself if you have the equipment and then propagate and distribute the yeast to anyone who would want it. i dont know how people feel about that morally given the emphasis on doing your own extractions on here though. it would likely still require some level of extraction.
 
Dirty T
#9 Posted : 11/17/2021 6:13:00 PM
Psilocybin and Psilocin are both water soluble so extractions wouldn't be an issue. You can do 100% efficient extractions with ethanol and water with a hot plate and very simplistic makeshift still. I've extracted several different ways and there is nothing needed to do so besides water or 70% ethanol. The extraction definitely falls within the scope of this site and is the easy hurdle. Shipping the culture is the sticking point.

You could also make psilowine with it as well. Making wine with psilocin producing yeast would give you a very active end product.
 
Ruffles
#10 Posted : 11/19/2021 10:56:30 PM
Are the metabolic pathways from (assumed precursor) tryptophan to hallucinogens actually known in any of the plant or fungi allies?

I know of limited suppositions based of artificial synthesis and known metabolites within the plants... but nothing like a full pathway describing the enzymatic reactions and their components.

Its highly unlikely that a single gene would make P. pastoris or S. cerevisae produce specific tryptamines, plus accumulate and not degrade it to get any usable yield after purification.

Also, to produce `psilins` like a recombinant unicellular organism would do, its easier to produce tons of cubensis mycelium using specific liquid medium to stimulate alkaloid production and extract out of that.
 
benzyme
Moderator | Skills: Analytical equipment, Chemical master expertExtreme Chemical expert | Skills: Analytical equipment, Chemical master expertChemical expert | Skills: Analytical equipment, Chemical master expertSenior Member | Skills: Analytical equipment, Chemical master expert
#11 Posted : 11/20/2021 1:16:55 AM
Ruffles wrote:
Its highly unlikely that a single gene would make P. pastoris or S. cerevisae produce specific tryptamines, plus accumulate and not degrade it to get any usable yield after purification.
.


Metabolic engineering of Saccharomyces cerevisiae for the de novo production of psilocybin and related tryptamine derivatives


The puzzle was largely solved in 2011, when s. seriviceae was biotransformed to produce morphinans. Simple tryptamines aren't much more involved. Both tyrosine and tryptophan metabolism genes are ubiquitously expressed and highly conserved across countless species.
"Nothing is true, everything is permitted." ~ hassan i sabbah
"Experiments are the only means of attaining knowledge at our disposal. The rest is poetry, imagination." -Max Planck
 
benzyme
Moderator | Skills: Analytical equipment, Chemical master expertExtreme Chemical expert | Skills: Analytical equipment, Chemical master expertChemical expert | Skills: Analytical equipment, Chemical master expertSenior Member | Skills: Analytical equipment, Chemical master expert
#12 Posted : 11/20/2021 1:31:25 AM
John.Brown wrote:
it costs like 1k usd

the price is per kb. Check the size of the genes in question.

also, this isn't something trivial like a plant extraction. there is certainly a learning curve. Certain precautions need to be taken into account, like using nuclease-free water and reagents, and a DNA-free work area and similarly cleaned tools. It's not unlike working with aseptic techniques, but screwing up a biotransformation is much easier. It can definitely be done at home, but it takes practice and considerable attention to detail.

I highly recommend reading Molecular Cloning: A Laboratory Manual, before attempting any genetic engineering experiments.
"Nothing is true, everything is permitted." ~ hassan i sabbah
"Experiments are the only means of attaining knowledge at our disposal. The rest is poetry, imagination." -Max Planck
 
Ruffles
#13 Posted : 11/27/2021 12:52:32 AM
benzyme wrote:
Metabolic engineering of Saccharomyces cerevisiae for the de novo production of psilocybin and related tryptamine derivatives


The puzzle was largely solved in 2011, when s. seriviceae was biotransformed to produce morphinans. Simple tryptamines aren't much more involved. Both tyrosine and tryptophan metabolism genes are ubiquitously expressed and highly conserved across countless species.


Thanks for that citation. Well, according to the paper it certainly takes more than one gene to get it working in S. cerevisae, plus it takes fine tuning of protein expression by fiddling with the promoters as CYPR overexpression messes with superoxide metabolism. Its an amazing proof of concept though.

I didn`t see any comparison of their top recombinant cerevisae to cubensis liquid mycelium production though... mass to mass comparison of yields or something like that. Still skeptical on it being a better factory than cubies.

I did find the concept of being able to produce other types of tryptamines extremely interesting, it could resolve synthesis of specific isomers in a way that classical chemical synthesis is not able to do.
 
roninsina
#14 Posted : 11/27/2021 4:51:02 AM
Sorry, I can’t recall where I read the recent article, but commercial researchers were quoted saying that these strains were so widely available for research that other avenues for patenting would have to be explored as it was already assumed the yeast was off the farm, so to speak.

Maybe it would be possible to acquire a culture (at least of the Psilo-yeast ) quasi-legally.
"We dance round in a ring and suppose,
while the secret sits in the middle and knows." Robert Frost

 
Dirty T
#15 Posted : 11/29/2021 5:16:40 PM
This thread got me thinking. I think using yeast for nn production is a waste, MH is a hearty plant almost anyone can grow at home. I believe a good use for this type of genetically modified yeast would be 5meo production. 5meo producing yeast would be awesome.

I am also in agreement that yeast seems like an inefficient route to producing psilocybin. I produce 2 oz dry fruit per quart of spawn in 23 days start to finish. The total materials cost is less than a dollar per oz. and time invested is right at 1 hour per oz as long as I go bulk. I have 2 PCs for the sake of efficiency so I run 14 jars at once rather than 7 (I don't turn them sideways and I "Jack them up out of the water" with pp5 containers beneath the bottom tray.)
 
 
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