Bacterial Production of Opiates

[Nightsd]

I have recently been reading up on bacterial transformation, and an interesting thought crossed my head.

β-endorphin is a naturally produced protein that agonizes μ-opioid receptors.

What do you think would happen if someone took the gene that synthesizes β-endorphin, and used that gene to make custom plasmid DNA with the β-endorphin production gene loaded on to it? I don't know exactly how plasmids are synthetically produced, but I do know that humans can synthetically manufacture them and you can order plasmids of your own specifications online. Anyways, if you took a bottle of these plasmids, and introduced them into heat-shocked bacteria (similar to the pGLO experiment that every bio major in college does), would the bacteria take up these plasmids, and then start producing β-endorphins? After bacteria have produced the β-endorphin, you could easily use a centrifuge to separate the β-endorphin from the bacteria.

So what do you guys think? I have no idea whether it would work or not, let me know what you think!

As a side note, β-endorphin is not permeable through the blood-brain barrier, but it's an interesting thought nonetheless. Maybe there is another type of naturally occurring opiate protein that is permeable through the blood brain barrier? I can think of an interesting lab experiment you could try on...rats...yeah, rats would work by the way, β-endorphin is approximately 80x more potent than morphine.

 

[sekio]

As you yourself said, beta-endorphin doesn't cross BBB and only produces peripheral effects.

Sure, you can produce 'fun' opioid peptides but even then those are best done on a peptide synthesis machine because they often use freaky amino acids (i'm looking here at dermorphin etc which do cross BBB).

Also, peptides are often broken down very rapidly by peptidases, making them unfeasible as long-lasting drugs unless modified or of special structures.

 

[Nightsd]

Thanks!

Apparently dermorphin also requires a post-translational modification after synthesis. Can this modification be synthetically induced in a peptide synthesis machine, and in modified bacteria?

Also, could you use a protease inhibitor to protect dermorphin & co. from degradation?

 

[sekio]

Can this modification be synthetically induced in a peptide synthesis machine,

Doesn't need to be, the peptide synthesis process doesn't care, it just uses chemical catalysts to link amino acids together like beads on a string as you feed them in one after another. Dermorphin is produced in the lab by scientists who have access to the freaky "wrong handed" amino acids. On the other hand bacteria lack the enzymatic machinery to flip amino acids around so unless you encoded them too (and at this point you are best off cloning frog skin cells) or provide them in the natural environment, they can't produce protiens like dermorphin.

More complex modifications would require either secondary processing after the peptide was made or really freaky enzymes.
.

 

[Hammilton]

Ah, that's post translation modification. You can't have post translation-modification after synthesis because if you were synthesizing it, there would be no translation because you're not starting from DNA, you're starting from amino acids. When you synthesize it, when you're done synthesizing it, you're done. With dermorphin instead of using L-alanine you'd be using D-alanine so there'd be no need to make any changes.

I'm no expert in peptide synthesis, but it doesn't seem like that'd pose any special challenges, it's no like it has some substantial modification, it's just a different isomer.

To do it bacterially you'd have to get some sort of isomerase expressed as well and hope that it worked. Synthetically it shouldn't be an issue at all because you wouldn't be modifying anything.

 

[Deleted member 170540]

http://www.ncbi.nlm.nih.gov/pubmed/15009527

A chimeric opioid peptide with mixed mu agonist/delta antagonist properties.

Abstract
There is evidence to indicate that opioid compounds with mixed mu agonist/delta antagonist properties are analgesics with low propensity to produce tolerance and physical dependence. A chimeric peptide containing the potent and selective mu agonist H-Dmt-D-Arg-Phe-Lys-NH2 ([Dmt1]DALDA) (Dmt=2',6'-dimethyltyrosine) and the potent and selective delta antagonist H-Tyr-TicPsi[CH2-NH]Cha-Phe-OH (TICP[Psi]) (Cha=cyclohexylalanine), connected 'tail-to-tail' via a short linker, was synthesized using a combination of solid-phase and solution techniques. The resulting peptide, H-Dmt-->D-Arg-->Phe-->Lys-NH-CH2-CH2-NH-Phe<--Cha[NH-CH2]PsiTic<--Tyr-H, showed the expected mu agonist/delta antagonist profile in the guinea-pig ileum and mouse vas deferens assays. Its mu and delta receptor binding affinities were in the low nanomolar range, as determined in rat brain membrane binding assays.

Would this peptide cross the BBB? Sounds interesting, less tolerance development and dependence than with traditional mu-opioids...

 

[alkap555]

In general, peptides do NOT readily cross the BBB unless specifically mediated by carrier-transport. There is evidence supporting the intranasal delivery of peptides directly into the CSF, effectively bypassing the BBB (see this thread thread: http://www.bluelight.ru/vb/threads/...tor-Neuroscience-behind-premature-ejaculation).

regarding opioid peptides and the BBB:

http://www.aapsj.org/view.asp?art=aapsj080109

Edit: An interesting idea though..I have always thought that the richest man in the world would be the one that figures out how to replicate the biosynthesis of cocaine in Erythroxylum coca and develop bacterium that could do the same.

Think about it, all you would need is some fermenting glassware, an autoclave, a culture of the bacteria and the proper precursors (amino acids etc), and extract cocaine from the broth as a secondary metabolite! The cartels would vanish and cocaine production would become domestic.

....One day...