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Genetically engineered DMT Options
 
benzyme
#61 Posted : 5/9/2012 6:18:44 AM

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adorno wrote:
Why isn't every million dollar lab doing enzyme catalyzed reactions along these lines already? I mean, couldn't they replicate these things by kilogram and streamline so many other costs of research/production?


they already are, but for pharmaceuticals, and biopharmaceuticals. to justify getting the government grants for funding this sort of research, you have to show medicinal value....or warfare value (go figure).
"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
 

STS is a community for people interested in growing, preserving and researching botanical species, particularly those with remarkable therapeutic and/or psychoactive properties.
 
nen888
#62 Posted : 5/9/2012 6:22:25 AM
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..great flow-chart benzyme..!

why do the enzymes end up costing so much..is the replication/cloning of them extremely complicated..
pardon my ignorance..or is it just because that's what a limited number of companies can charge if they want?
like, is it expensive equipment, or patented technology going on here?
.
 
benzyme
#63 Posted : 5/9/2012 6:33:21 AM

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like any other technology, you're paying for research & development, purification, analysis, etc., as well as packaging and shipping.
but yea, the fact that they are niche products allows companies to command high prices
"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
 
crystalizer
#64 Posted : 2/13/2013 9:18:40 PM

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Hey! First of all THX for all this theoretical imput and for keeping this topic alive...

Reasearch is like Infundibulum says... sometimes it takes more time than expected. For insance i recieved in the beginning of 2011 a small project - 2 months maximum was planned. ... here we are two years later, with the particular project still not finished....



@Infundibulum
INMT and TDC shoud be tagged, yeah... isn't the simple his tag eough? purify it and end up with correct bands (SDS-PAGE)...? or do you really thinking about westernblotting it?


@ MR. M.S cell/molecular biology - whitebread, i agree with endlessness and Infundibulum. they both gave a very statisfying replies - i have nothing else to add...


@ adoro : NICE, very interesting input. when i find some more time, i will go through your thread.... i have to agree with benzyme and Infundibulum, enzyme purifications are off the table ...
i worked with tabacco in cell cutre... it wont work at home...you are going to contaminate everything before it even starts growing...
the bioreactor pipeline is also great, of course we will use one Pleased but FiRST we need some basic research, about which bacteria/enzyme is doing the job, at which conditios etc. and then we can think about pipelining it...


BTW - you all convinced me of trying it in yeast (if i ever try it...) - even if the colis would grow faster... for home usage yeast is more convinient... some nice stock in the freezer, put it in culture and harvest couple days later... it will be still faster than trying to find some mhrb ...


"It is not the strongest species that survive, nor the most intelligent, but the ones most responsive to change."
-NOT Darwin
 
closet-chemist1010
#65 Posted : 2/14/2013 1:41:26 AM

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hey, has nobody seen Rhodiums chemistry archive? after the hive, and rhodiums personal site went down, erowid launched archived version of each, if im not mistaken there is a detailed chemical synthesis of dmt in there.
 
benzyme
#66 Posted : 2/14/2013 1:49:43 AM

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nah, nobody's ever seen it. news to me.

you can't do synths better than the microbes.
enzymes are substrate-specific and won't produce cyclized
byproducts.
"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
 
centromere
#67 Posted : 4/12/2013 8:17:05 PM
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benzyme wrote:
whitebread was merely suggesting what infundibulum and I had already pointed out...it's not a
cheap nor practical project. Paying for all the equipment and reagents is half of it. The other half is getting the proteins (enzymes) of interest to express in vitro. Of course, the organism should already express proteins for tryptophan metabolism (and many bacterial and yeast cells do); keep in mind, these proteins are substrate specific, so you can't toss in any molecule, like Indole-3-Acetic acid, into a vat of cells expressing INMT, and expect it to get converted to DMT. it doesn't work like that.



the straightforward route is

tryptophan -4.1.1.28-> tryptamine + SAM -2.1.1.49-> NMT + SAM -2.1.1.49-> DMT



Sorry to resurrect this old thread, but the pathway map above is very intriguing. According to the map, it is entirely possible for yeast to make N,N-DMT without any genetic modifications. Of course, the quantities of DMT are likely quite low, if any.

According to the map, tryptamine is used as a precursor in the production of N-hydroxy-tryptamine and indole-3-acetaldehyde. What if one were to knock out the genes responsible for those products (1.1.4.1.3- and 1.4.3.22/1.4.3.4)? If those knockouts do not prove to be lethal, it would elevate tryptamine levels in the cell allowing it to be used exclusively by 2.1.1.49.

This is highly speculative, but I'm interested in the community's take on it.
 
embracethevoid
#68 Posted : 4/13/2013 4:47:50 PM

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If yeast can do it, I don't see why it would not be possible to skip the whole GM doodad and just go for plain selection and culturing of it?

1.) Find test for DMT
2.) Kill all the yeast that do not produce DMT, or if you are more merciful, separate higher DMT producing strains from there
3.) 10,000 generations later, ???
4.) Profit!!!!


Worked for marijuana, didn't it? I hear THC content started off like 4% and now it's hitting 30% just due to careful breeding.


Of course, this tek could be adapted towards acquiring HP_DMT. I'm off to eat some babies now, will report back.
 
SKA
#69 Posted : 4/14/2013 2:44:17 PM
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Fascinating. Never knew yeast DID make DMT. It should be possible but time consuming.
I mean...How long did cultivators take to increase Marijuana's THC content from 4% to 30%?


I guess it would be more effective & less time consuming to grow Phalaris strains, take measurements
& separate and cultivate only those specimens with the highest DMT contents. This could be done with
any fast growing DMT plant that can survive easily in many different climates, so the result would be
fast-growing, Frost/Moisture/Darkness-resistant plants/trees who's DMT content rises with every new
generation. Acacias, Phalaris, Desmanthus....All of these are probably better candidates for this
experiment than yeast, given their already significant DMT contents.

But IF you could get it done with Yeast, it would yield the easiest growing DMT source.
Don't let this marvelous topic catch dust. Any news ?
 
centromere
#70 Posted : 4/14/2013 5:25:14 PM
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SKA wrote:
I guess it would be more effective & less time consuming to grow Phalaris strains, take measurements
& separate and cultivate only those specimens with the highest DMT contents. This could be done with
any fast growing DMT plant that can survive easily in many different climates, so the result would be
fast-growing, Frost/Moisture/Darkness-resistant plants/trees who's DMT content rises with every new
generation. Acacias, Phalaris, Desmanthus....All of these are probably better candidates for this
experiment than yeast, given their already significant DMT contents.


I disagree with you that phalaris is less time consuming. The turnover rate for yeast is far higher -- they go exponential in a matter of hours.

Assuming that yeast actually do produce DMT (we're not 100% sure), we would need a way to test for DMT presence. That is NOT trivial, considering that there are tens of thousands of different chemicals inside yeast. Making a simple test-kit to identify DMT (and not anything else in the cell) would be incredibly difficult.

I guess the first step would be to figure out an easy way to test for the presence of DMT, and then test the yeast. If you get a non-zero result, then you could use that as a reference point for future generations. But if you get a zero result, you can't proceed because you won't have any basis to select for which yeast produce more.
 
adam
#71 Posted : 8/15/2014 6:43:58 AM

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I know this thread has been resting for a while but, I thought this might be the right place to post this.

recombinant dna technology
 
Adjhart
#72 Posted : 8/16/2014 11:10:14 PM

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Wow.



It took me an hour and a half to read 3 pages and I feel like I just got beat up. Surprised Everything was so alien to me; I barely managed, somehow.

The expertise that shines through some of these posts is incredible. Shocked

It's truly an honor to be in the company of our scientists here at the Nex.

Thumbs up
 
starway6
#73 Posted : 8/18/2014 9:42:24 PM

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@Infundubulum: Yeah proof of principle would be great for beginning Anyways there is no way of spending MY money on this The effort... well, people put a lot of effort in different kind of (crazy) things, consider it as a hobby of mine . The overall achivement - well considering the tiny tiny quantities of DMT present in natural sources and a long way of chemical synthesis - there should be an alternative way to acess great quantities of DMT, which would be neccessary for lets say reasearch - if one day somehow it should be legalized...
Coming to the smell. Well, I worked with crazy ass bacterial and fungal cultures, THEY REALLY SMELLED LIKE SHIT. Thus I consider colis and yeasts as relative mild. Thinking about it, you are maybe right, havent worked since a long time with yeast, so the actually might smell better. BUT considering smell: when I read, that people extract their DMT with naphtha - geeeeee - my hair stands up like an urchin. I would't even lighten my BBQ with that nasty smelling crap...


What is cleaner extraction chem than napatha in your opinion?Wut?
 
benzyme
#74 Posted : 8/19/2014 1:19:36 AM

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naphtha smells like roses compared to some chemicals used in biotech, like b-mercaptoethanol; and it's fairly innocuous, really, when you're used to extracting with xylene and dcm (which only require one pull).
"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
 
darklordsson
#75 Posted : 8/19/2014 4:29:50 AM

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Sounds like im going to get some xyleneBig grin

Great topic tho guys im in aw at the knowledge here too as well. Many thanks benzyme, you know the dance man, hats off to you!Thumbs up

 
benzyme
#76 Posted : 8/19/2014 5:00:40 AM

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thanks. Big grin

the reason people favor naphtha is it works well for freeze precipitating out the alkaloids, xylene does not (but you can salt it out with fumaric, tartaric, maleic acid, etc).
the other thng is naphtha actually doesn't fill a dwelling with its aroma without a fan blowing over it. xylene will, without a fan.

dcm is old-school, very effective and not so eco-friendly, my favorite. food-grade dcm Thumbs up
"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
 
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