View Full Version : Converting morphine to hydromorphone w/ bacteria
dugwylor
12-19-2008, 08:39 AM
Ran across this on Wikipedia:
"Some bacteria have been shown to be able to turn morphine into hydromorphone. As reported in the July 1993 issue of Applied Environmental Bacteriology, the bacterium Pseudomonas putida, serotype M10 produces a naturally occurring NADH-dependent morphinone reductase which can work on unsaturated 7,8 bonds—with result that when these bacteria are living in an aqueous solution containing morphine, significant amounts of hydromophone form as it is an intermediary metabolite in this process; the same goes for codeine being turned into hydrocodone. At this time no information has been published indicating whether or not this process can change dihydromorphine into metopon or acetylated morphine derivatives into the respective ketones of the acetylmorphone series."
Interesting stuff. If someone were to find a way to access this organism, grow it in an easily-available morphine/codeine solution for X amount of time, and then filter-sterilize or just saturate w/ EtOH, that individual would have a much more interesting hydromorphone/hydrocodone solution. Not sure how tricky it would be to access this organism (might need bio connects, which a particular hypothetical individual might have). P. putida is interesting in that it's the very first patented organism in the world. It's used for a number of things, including breaking down polystyrene foam into usable plastic.
Anyone heard of this or see any problems w/ this? There's a paper on the subject (link on Wikipedia) if anyone is into M/CB.
Narkotikon
12-19-2008, 01:48 PM
I've read something on Wiki about this too, but I think it was bacteria that turned hydrocodone into hydromorphone, which I can see, because that's what happens when your body metabolizes hydrocodone. I didn't know it could work with morphine too though. Interesting.
Synack
12-19-2008, 02:03 PM
codiene into hydrocodone and morphine into hydromorphone .. pretty cool - Anyone have this article on hand?
edit: here's the article http://www.pubmedcentral.nih.gov/picrender.fcgi?artid=167664&blobtype=pdf
...Anyone heard of this or see any problems w/ this? There's a paper on the subject (link on Wikipedia) if anyone is into M/CB.
The problem is that you need morphine, and it would be far easier to just convert it to heroin
or analog compound by traditional methods.
Even when produced by microorganisms, the desired compound must be isolated and purified so odors etc
from solvents must still be considered. Also, you will need much more equipment.
Cloning genes etc into microorganisms to produce biopharmaceuticals is what I do for work,
in a research lab not large-scale production.
Setting up the bioreactor for optimal production of the target compound is a lot of work,
but once that is accomplished its pretty simple to maintain assuming the target
compound is excreted by the bacteria so you don't need to lyse cells etc.
(in the article posted, the compound is excreted)
If you have any questions about terminology or things like that, I can answer them.
I don't know where to get P. putida M10 (haven't looked), but if you want to get some and the necessary
equipment you should have a business name as you will almost always need to
establish an account. Its just a formality, not hard to do.
dugwylor
12-21-2008, 01:52 PM
The problem is that you need morphine, and it would be far easier to just convert it to heroin
or analog compound by traditional methods.
One reason some people might find the method described in this thread interesting, though, is that from what I've seen, the route to diacetylmorphine always requires acetic anhydride -- something which most people don't have much access to. It's not all that difficult to arrive at a relatively pure morphine product from available sources, though. Once you're to that point, there's been some talk about conversion to monoacetylmorphine using simple glacial acetic acid... what's the particular downside to going this route? I've heard there's a potential issue of toxicity. Mayo, know much about this? Some people may simply wish to explore various interesting options with limited knowledge and moderate access to lab supplies.
Hi dugwylor.
Yes, it is interesting using microorganisms to produce pharmaceuticals.
I can't speak about how to make drugs, sorry.
Getting small amounts of acetic anhydride is not nearly as difficult as many people think.
It is available in 'kits', thats all I'll say about that.
Other less-watched anhydrides can be used, which is why I mentioned 'analog compounds'.
I didn't catch-on to your mention of 'easily-available morphine/codeine solution' but
now I understand what you were referring to. Just add that and nutrients to the
bacteria and get your good stuff, right? I'm sure you know it is far more complicated than that.
Unfortunately, the bacteria will be excreting many things in addition to the desired
product. In fact, there will be so much other crap in there, getting a pure compound
from this mixture will require a work-up that makes extracting M and synthing H or analog
seem like making a cup of coffee.
Just the cost of the bioreactor with the necessary probe-type sensors, stirrer and motor,
monitoring software, etc. etc. would buy you a truckload of pods.
But who knows, maybe you could just throw all the stuff in a 5 gallon bucket wrapped in a couple heating pads,
wait 24 hours and start extracting the goodies!
dugwylor
12-24-2008, 08:54 AM
Hi dugwylor.
Yes, it is interesting using microorganisms to produce pharmaceuticals.
I can't speak about how to make drugs, sorry.
Even done entirely from a curiosity perspective? Well, thanks for going as far as you may, at the very least. Don't want any trouble to stir. I'm still not entirely certain of the rules involved here; for example, what would be the difference between referring to the very well-known process of converting O to H, and discussing lesser-known processes published in scientific journals? MAM is hardly "new," but from the small amount of information available it's difficult to discern why it isn't being produced far more regularly than DAM...
Getting small amounts of acetic anhydride is not nearly as difficult as many people think.
It is available in 'kits', thats all I'll say about that. Other less-watched anhydrides can be used, which is why I mentioned 'analog compounds'.
Both are extremely interesting. However, one would think "kits" would still be well-monitored by LE? Also, I wasn't aware that substitutes could be used; you'd think a curious amateur chemist (with focus on the amateur) might run some risks with substitutions, regardless of access to materials. Discussion with chemists in person might not also be entirely feasible for any such hypothetical person, though doctoral microbiologists with at least strong org. chemistry knowledge may.
I didn't catch-on to your mention of 'easily-available morphine/codeine solution' but
now I understand what you were referring to. Just add that and nutrients to the
bacteria and get your good stuff, right? I'm sure you know it is far more complicated than that.
Unfortunately, the bacteria will be excreting many things in addition to the desired
product. In fact, there will be so much other crap in there, getting a pure compound
from this mixture will require a work-up that makes extracting M and synthing H or analog
seem like making a cup of coffee.
Just the cost of the bioreactor with the necessary probe-type sensors, stirrer and motor,
monitoring software, etc. etc. would buy you a truckload of pods.
But who knows, maybe you could just throw all the stuff in a 5 gallon bucket wrapped in a couple heating pads,
wait 24 hours and start extracting the goodies!
Naive hopes are still hopes, and they become satisfied more often than one would think! Well, here's to interesting ideas.
I wasn't joking about this. To do a one-off rather than perpetual type of operation, just meeting growth requirements.
But who knows, maybe you could just throw all the stuff in a 5 gallon bucket wrapped in a couple heating pads,
wait 24 hours and start extracting the goodies!
...I'm still not entirely certain of the rules involved here; for example, what would be the difference between referring to the very well-known process of converting O to H, and discussing lesser-known processes published in scientific journals?
I'm not entirely sure what the rules are here.
I'll talk about harm-reduction related chemistry but no synth, not even from microorganisms. Not in a 'how-to' manner anyway.
If someone wants to talk about some science topic, and they are serious about the discussion,
I'll go for that most of the time.
...there's been some talk about conversion to monoacetylmorphine using simple glacial acetic acid... what's the particular downside to going this route? I've heard there's a potential issue of toxicity. Mayo, know much about this? Some people may simply wish to explore various interesting options with limited knowledge and moderate access to lab supplies.
There's been some talk? ...lol.Ok. Yes, some monoacetylmorphine will be produced using GAA, and a catalyst,
but there are reasons this is not the preferred route. If you knew some chemistry
you would know why. No offense. This is very simple stuff. But the answer is yes...I'll talk organic synth with other chemists, because
then it would be an actual conversation..lol..but again, not here. Thats just my position, not meaning to say those are the rules etc.
I understand your curiosity, deeply. I'm sure others will be glad to discuss these things with you. I'll be glad to talk some science with you too.
Tea Time
12-24-2008, 04:36 PM
I seem to remember someone mentioning this here before, but it is very cool. Not very practical, but it is cool that it is possible.
After reading my last response, it sounds kind of dickish. I did not mean it to be that way.
Sometimes I try to be amusing and it just never sounds right.
If you have any questions dugwylor, feel free to ask. I may decline to answer, while someone else may reply.
But no harm in asking I would think.
Also, I don't know what you meant by 6-mam synthesis and toxicity.
6-mam itself is not considered toxic (aside from being an opioid) but toxic compounds would need
to be removed during work-up.
I have a question....would it even be possible for average joe to get gene-spliced morphine producing bacterium? It seems to me that if you were to make a guide for this, it would go something like:
"Step 1. Go to _____labaratories
Step 2. Find security guard named Jose. Talk about weather and then bribe him...."
etc etc.
Like it's not something you can just MAKE, you'd have to get it from another source of that bacteria, almost like yeast, right? Oh man, if they could splice the morphine producing gene into YEAST......our problems would be over!
Yes, it is 'made' as you said. The gene itself has a source, say a plant, and is 'cut out' using enzymes.
Then many copies of the gene are made, or cloned, using one of a few methods, like plasmid DNA, which can then be inserted into the chromosomal bacterial DNA usually along with a promoter (another segment of DNA).
These are the basic steps to 'make' the transgenic bacteria. You don't need a lab to do it, just the lab equipment.
Does that answer your question? I tried not to use 'technical' words, or at least explain them, and be as brief and basic as possible.
Oh, as far as making morphine. Several genes are utilized by P. somniferum in a multistep process. I haven't looked into whether this has been done, but I doubt it considering how easy morphine is obtained 'naturally'. But it may have.
One thing: Genes don't make drugs, if thats what anyone was thinking. Genes are coding segments of DNA, which are transcribed into
RNA, which is translated into a protien, in this case an enzyme. The enzyme acts on a molecule that is being modified in some way to produce the 'drug'. This is probably obvious to everyone, just making sure.
And one last thing..lol.The strain of bacteria P. putida M10 was not 'made', it already has a gene that codes for an enzyme which acts on morphine.
the bacterium Pseudomonas putida, serotype M10 produces a naturally occurring NADH-dependent morphinone reductase
I have a question....would it even be possible for average joe to get gene-spliced morphine producing bacterium? It seems to me that if you were to make a guide for this, it would go something like:
"Step 1. Go to _____labaratories
Step 2. Find security guard named Jose. Talk about weather and then bribe him...."
etc etc.
Like it's not something you can just MAKE, you'd have to get it from another source of that bacteria, almost like yeast, right? Oh man, if they could splice the morphine producing gene into YEAST......our problems would be over!
... You don't need a lab to do it, just the lab equipment...
Ha! I forgot to finish that thought. That sounds so ridiculous. Thats what happens at 4am and can't sleep.
No sophisticated equipment is really needed. The electroporator should be by far the single most expensive item, the cheapest costing about $1,200.00 new. Next would be an incubator, which can be obtained for a few hundred dollars. Mostly beakers, flasks and culture dishes.
Typical reagents and growth media are needed of course. Reagents more specific to molecular biology will be needed. There are many items, but none are expensive.
What REALLY makes cloning possible for 'average joe' is that now you can buy complete cloning kits and transformation kits. These are supposed to contain all the many necessary enzymes, probe, vector and eukaryotic expression system. In the lab we design our own 'experiments' and assemble the necessary items. This is a huge amount of work and requires extensive knowledge of molecular biology to be done well.
All 'average joe' really needs to be able to do is follow the kit instructions.
But IMO to have a decent chance of a success he will need to learn the basics of cloning and transformation.
I'd estimate joe could produce his bacteria for at most $4,000.00 including the electroporator. Skip the cost of this item by using calcium chloride to induce competency, but check with the kit instructions first.
Didn't mean to make it sound like i was trying to find out how to do this for myself....it's way beyond my skill and knowledge.
I just think it's interesting that some day there might be a strain of bacteria that one could get a sample of, take it home and grow their little baby bacterium into a big healthy bacterium, all the while harvesting morphine from it. I mean, if there was a bacteria that could take say, agar, heat, and moisture and turn it into morphine.....it'd be impossible to control, wouldn't it? What once took months during the proper season, could be done in a 25 gallon heated tank and some potatoes, in someones basement, closet, or even a "seed" culture in the trunk of a car. And then once one person gets a stable sample....they can give a culture to someone else who can start their OWN sample, and make it grow exponentially.
I'm sure it's not as simple as it sounds, but my point is this: if someone already HAD a sample of E.coli that produced morphine, wouldn't it be much, MUCH easier? Or is this e.coli chimeric and therefore sterile (can bacteria BE sterile? would that make it die out almost instantly?) ? And then there's scrubbing the final "harvest" but that can't be *that* hard can it? (or can it? i really don't know thats why i'm asking:D)
Of course this could also be very dangerous, since it IS still E.coli, and possibly deadly, correct? Imagine an outbreak of morphine-producing e.coli....OR on the flipside, a benign bacterium that produces morphine and lives in your intestines....hell, there might already BE bacteria like that in our own bodies. the medication pumps of the future, hah.
dugwylor
12-30-2008, 08:50 AM
No sophisticated equipment is really needed. The electroporator should be by far the single most expensive item, the cheapest costing about $1,200.00 new. Next would be an incubator, which can be obtained for a few hundred dollars. Mostly beakers, flasks and culture dishes.
Typical reagents and growth media are needed of course. Reagents more specific to molecular biology will be needed. There are many items, but none are expensive.
What REALLY makes cloning possible for 'average joe' is that now you can buy complete cloning kits and transformation kits. These are supposed to contain all the many necessary enzymes, probe, vector and eukaryotic expression system. In the lab we design our own 'experiments' and assemble the necessary items. This is a huge amount of work and requires extensive knowledge of molecular biology to be done well.
Let's take a particular hypothetical average Joe; let's say he has access to all of the equipment you name, and while having almost no competency in molecular biology, controls ordering for a lab and can work closely with a possibly-willing molecular biologist. It's actually not such an unusual situation, when you think about it; this could be one reason why such a person might be more seriously inclined toward pursuing this method. With those hypotheticals in mind, how difficult do you think it would be to obtain P. putida M10, and roughly how long would the protocol take to complete, from start to finish? Oh, and would you happen to have any idea what sort of media you'd grow it on? It makes an interesting hypothetical.
Crispkreem
01-01-2009, 04:48 PM
exteremely interesting,my curiosity is piqued i would to dicuss this further with anyone else interested
Hell, I'll discuss it, but I don't really know much about it. the one thing i really am curious is would it be POSSIBLE to make a type of bacteria that produces morphine, that could be stable enough to grow at home.
MissingOxy
01-28-2009, 09:40 AM
On the acetic anhydride front, acetyl chloride will do as good or better a job..With acetic anhydride, water is produced by the reaction. Using acetyl chloride produces HCl (hydrochloric acid) which vaporized out of the soltion. Warning: HCl gas is corrosive, so you'd better have a great ventilation system.
MissingOxy
Just a quick question to those who might know, its a little off the subject matter, but similar.
Would it be possible to splice the morphine producing genes out of p.somniferum into other types of plants, maybe easier to grow plants that also produce white latex substances as a matter of course? (I'm thinking dandelion weeds here...?) If so, one would literally have a weed that produces alkoloids without having to worry about the negative consequences of growing p.somniferum in public. Additional benefit is that it would literally be as easy to grow as weeds...
Would something like this be possible to do or easy to do?
Thanks for any responses to this non-scientist but very interested in science kind of guy...
On the acetic anhydride front, acetyl chloride will do as good or better a job..With acetic anhydride, water is produced by the reaction. Using acetyl chloride produces HCl (hydrochloric acid) which vaporized out of the soltion. Warning: HCl gas is corrosive, so you'd better have a great ventilation system.
MissingOxy
What would this give? MAM or diacetylmorph?
Just a quick question to those who might know, its a little off the subject matter, but similar.
Would it be possible to splice the morphine producing genes out of p.somniferum into other types of plants, maybe easier to grow plants that also produce white latex substances as a matter of course? (I'm thinking dandelion weeds here...?) If so, one would literally have a weed that produces alkoloids without having to worry about the negative consequences of growing p.somniferum in public. Additional benefit is that it would literally be as easy to grow as weeds...
Would something like this be possible to do or easy to do?
Thanks for any responses to this non-scientist but very interested in science kind of guy...
That's sort of what we were talking about, but i wanted to take it one step further: just put it into a simple bacteria, so you don't need anything other than something like agar and water. Is it possible? Yes, but it's not something that you could just do as a weekend project or something. And I think there are some hardcore laws about letting a mutated plant be released into the wild, but i'm not sure.
MissingOxy
01-30-2009, 01:06 AM
Diacetylmorphine is the result.
(1) morphine + (2) acetyl chloride <---> (1) diacetylmorphine + (2) HCl
-or-
(1) morphine + (1) acetyl chloride <---> (1) monoacetylmorphine + (1) HCl
(1) monoacetylmorphine + (1) acetyl chloride <---> (1) diacetylmorphone + (1) HCl
As the HCl is a gas, it leaves the reaction vessel forcing the the equation to the right. So basically 1 mole of morphine + 2 moles of acetyl chloride yields 1 mole of diacetylmorphone + 2 moles of HCl. Use a small excess of acetyl chloride, and the yield is virtually 100% by molar quantity. By weight, it's even better! The moecular weight of morphine is 285.5, diacetylmorphine is 369.41. If you start with 10 grams of morphine, the yield of diacetylmorphone would be (10*369.41/285.5)=12.93 grams (129% yield by weight). Wash the base product with water to remove any excess acetic acid remaining in the final product, or use sodium bicarbonate to neturalize any excess acid then wash with water to remove the water-soluble byproduct (sodium acetate & sodium chloride).
While the process is relatively simple, unless you're used to handling corrosives :deal2:it might not be a great first chemistry experiment.
MissingOxy
nhop, inserting genes into plant DNA is much more difficult than with bacteria.
Is what you're describing possible - maybe, by very experienced people, and would cost a fortune.
Easy? - ha..no way..in a word: forgetaboutit..
The two ways I know of to insert genes into plant DNA is using a specific plant virus (viral vector),
and the 'shotgun' method - shooting tiny gene-coated spheres into the plant cells.
Actually there is one newer way I don't recall the details of right now, but is still not easy.
Indy, I believe any genetically modified organism is classified as a 'level 2 organism'
at the least, which is regulated by the FDA and technically requires authorization to be taken out of the lab,
not to mention released into the environment.
But oversight is lax to say the least, basically relying on the professionalism and conscience of scientists,
so god knows whats been released.
Science..conscience..con-science...
Would it be possible to splice the morphine producing genes out of p.somniferum into other types of plants, maybe easier to grow plants that also produce white latex substances as a matter of course? (I'm thinking dandelion weeds here...?) If so, one would literally have a weed that produces alkoloids without having to worry about the negative consequences of growing p.somniferum in public. Additional benefit is that it would literally be as easy to grow as weeds...
Would something like this be possible to do or easy to do?
Yeah, i figured it'd be something like that. I was thinking back to that one agricultural research facility where the whole thing is underground and they're really careful about letting mutated plants out. It's like an underground farm, can't remember what it's called though.
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