Botulin extraction?

[akinrog]

Dear Forumites,

While searching the net I found the following patent (http://www.pharmcast.com/Patents/Yr2003/Jan2003/011403/6506399_Botulinism011403.htm) which IMHO gives hints regarding the extraction of (pure?) Botulin (which causes botulism disease (food poisoning).

Although the patent contained in this link is not directly related to extraction of botulin, there are some hints regarding a possible extraction by forming Zinc complexes of botulin.

The important part (again IMHO), is the part that states botulin forms a complex with Zn++ ions. I can readily assume that the zinc-botilum complex precipates enabling the chemist to separate the pure protein.

Any ideas? (I mean is it possible to extract the protein from a slurry, or anybody else know how this stuff is produced/extracted?)

P.S. Since botulin is not a genunie CW (for it is biologically obtained) nor it is a genuine BW (for it's a protein meaning it originates from biological sources), I could not post this under the existing threads. However, please move this post to relevant thread (if any) if you find creating a new thread for this subject is inconvenient.

P.S.2 The toxicological data for this protein is very interesting (ranging from mg order to picogram order :eek: ) according to this link (http://hazard.com/msds/tox/gw.cgi?query=botulin&whole=partial&start=0). But natural product's toxicological range is almost at picogram level.

[akinrog]

Hi,

I found following article (http://www.pubmedcentral.nih.gov/picrender.fcgi?artid=377283&action=stream&blobtype=pdf) entitled with the title of this post.

This article gives a method for producing botulinum toxin by the bacteria. I am now searching for method for isolating the pure toxin. Regards


IMPORTANT EDIT : Unfortunately when I searched deeper into this issue, I noticed C. Perfringens species do not produce botulism toxin but is a bacteria responsible for gas gangrene. Sorry for this inconvenience. However, we may readily assume that C. Botulinum may also be cultivated in this manner.

[nbk2000]

In Uncle Festers "Silent Death, 2nd ed." on the FTP, you'll find some references to botulism cultivation. But they're rather incomplete. Rather, use the provided citations to find further information in the relevant journals.

I found "Journal of Infectious Diseases" to be especially informative, as the early ones (pre-40's) to be very do-able on an experimentialist scale. Things like crystallizing typhoid toxin, cultivating gas gangrene, and all those other neat-o things you can do with germs. :D

[akinrog]

Things like crystallizing typhoid toxin, :D

I was looking for something like above. I am now in search of a method to precipitate the botulin protein from the broth.

Some theoric approaches may be like this:
First filtering the cultivated broth (which contains little germs) so as to remove suspended solids.
Then finding a method (which IMHO involves adding a Zn++ ion) to the filtrate to precipitate the protein. However, it is necessary to determine which salt of Zn is suitable for this process.
And then how to recover Botulin from Botulin-Zinc complex. :(

[akinrog]

In Uncle Festers "Silent Death, 2nd ed." on the FTP, you'll find some references ......

Argh! Fool me! I didn't listen to (or more properly read) what you wrote. I now actually scrutinized relevant section in Silent Death and it really contains a pretty good method (which involves precipitation of the toxin protein with acid and not even related to my foolish (or Kewlish) assumptions).

I think I am a fixed minded guy :(. Sorry for the trouble I created. Anyway thank you for your hint.