JOESTALIN (Newbee / Eraser)
02-26-01 10:34
No 175537
      Biosynthetic Pathway  Bookmark   

I have recently thought about developing a biosynthetic pathway for creating amphetamine in a recombinant E. coli.  This involves uses that process that E. coli already possesses for sythesizing Phenylalanine and using a gene encoding for the protein Benzylcarboxyhydrogenase.  This protein will take any aromatic carboxcylic acid/amino acid and reduce it using NADH to a methyl group.  By utilizing a exocellular transport sequence for the protein.  One could isolate the amphetamine product by simply pelleting the cells, evaporation, and using simple acid base extraction.  Now here's where the great part comes in.  All it would take to power this reaction is sugar, water, some salt, and a single cell.  It could be made in any quantity, placed anywhere near room temperature or warmer in the dark, no expensive reagents or glassware, and very little incriminating experience.  And best of all no experience necessary.  Tell me what you guys think.

Your Happy Little Dictator JOESTALIN

I want my baby back ribs, bbq sauce
(Hive Addict)
02-26-01 16:03
No 175574
      Re: Biosynthetic Pathway  Bookmark   

Go for it!!
Then quietly distribute the little guys far and wide.
This IS the solution to the war on drugs as we know it!!
Make everything as easy to produce as alcohol and "they" will never be able to stop it!!!!

Do Your Part To Win The War
02-28-01 00:00
No 175810
      Re: Biosynthetic Pathway  Bookmark   

What type of organism is the Benzylcarboxyhydrogenase in?  What is the GenBank number for it?  Refs?
(Newbee / Eraser)
03-03-01 05:33
No 176250
      Re: Biosynthetic Pathway  Bookmark   

It is a new biotechnology.  it is an engineered protein created by a researcher in a nearby lab, he is still waiting patenting, so he hasn't released it to Gen Bank. It is a genetic mutation of a chymotrypsin, shearly by accident trying to mutate it into a protease that would just cleave at Phenylalanine instead of it plus tyrosine and tryptophan.  Anyway, you won't find it in gene bank and I really had to kiss ass just to get a template.  But by the end of the summer we'll see if anything happens.

Your Happy Little Dictator JOESTALIN

I want my baby back ribs, bbq sauce
03-15-01 17:03
No 178696
      Re: Biosynthetic Pathway  Bookmark   

Is the enzyme availible comerically? 

Is there an article written about it?  Any literature?  Was there a letter to an editor even?  Please give more info...
03-15-01 20:25
No 178733
      Re: Biosynthetic Pathway  Bookmark   

JOESTALIN: Stop posting this nonsense!
04-02-01 08:40
No 181558
      Re: Biosynthetic Pathway  Bookmark   

Well...I've waited a month hoping you'd share more info.  So here's a BUMP.

And what is the substrate specificity for this enzyme?  Does it work on olefins?  Safrole???
I also think you underestimate the difficulty in separating the goodies from the ecoli goop.
04-02-01 13:26
No 181594
      Re: Biosynthetic Pathway  Bookmark   

As I wrote: it's completely nonsense.
(Hive Bee)
04-02-01 21:30
No 181685
      Re: Biosynthetic Pathway  Bookmark   

JOESTALIN may be full of shit here but there are bio-pathways to methamphetamine and amphetamine. 

How do I know this?  Well, methamphetamine and amphetamine both occur naturally in Acacic rigudula.  Since plants create all their goodies via enzymes I conclude that there is some enzyme that makes meth.  From what I don't know, but phenyalanine is a good guess.  PERHAPS, the meth is a result of a bacterial symbosis, but still there is an enzyme.

The most fucked up path I can imagine is that the meth is a result of accidential reduction or oxidation of another molecule  being synthesized imperfectly.  For example the synthesis of Molecule A is done via an enzyme B that results in some free engery that acts on molecule C which turns into meth.  (Not very scientific am I)

If you accept that the presence of meth and amphetamine reported in the paper on my website is indeed correct the possiblity of some enzyme exsisting becomes more plausible.

(Hive Bee)
04-27-01 09:35
No 187244
      Re: Biosynthetic Pathway  Bookmark   

I've posted about substituted ampehetamies in this plant and a closely related species, A. berlandieri a number of times and have looked closely at related pathways to try to determine where the original skeleton is coming ideas so far are:
1)Some sort of reduction step from ephedra alkaloids (which are found in other plants, but unfortunately not detected in these...not sure if the extraction procedure they used would pull these out, but I think they should have shown up if they were probably not this)
2)An enzyme with a decarboxylase/methyltransferase combined could use PLP to decarboxylate phenylalanine (or tyramine) and then use the delocalized lone pair to use the alpha-carbon as a nucleophile for attack on SAM or THF etc...I like this one simply because PLP-dependant enzymes kick ass  blush
3)Transaminase activity to form amphetamine skeleton from a related phenyl-2-propanone (similar to reductive amination, only the transaminase uses the amine version of PLP (there it is again  wink), PMP as the amine donor, water is lost. This type of activity has been documented in Phe and Tyr, why not the P-2-Ps?
I do have a couple other putative enzyme mechs, but they lack the supporting related enzyme activities, and really don't sound as good...

Begin with the dissolution of superfluous matters
So that desire and consciousness are free