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-   -   [x-post] Enzyme deactivation (http://www.molecularstation.com/forum/chemistry-forum/44174-%5Bx-post%5D-enzyme-deactivation.html)

Andrea 11-13-2003 07:37 AM

[x-post] Enzyme deactivation
 
Hi everybody, here is the question:

I use an enzyme (a cellulase) to degrade a cellulose derivative.
At a certain point of the process I need to stop it by deactivating
the said enzyme.
I can't use Hg compound anyway.
I tried to deactivate the enzyme by high temperature, but my product
doesn't like very much heat.
I've just read somewhere about deactivation by high shear...!
What about?
Do you know other ways to deactivate the enzyme.
Have you any advice for me?
I should be very happy if you want to suggest me some web resource
about the matter.
Thanks in advance.

Andrea Mitarotonda

Steve Turner 11-13-2003 11:31 PM

[x-post] Enzyme deactivation
 
[Only registered and activated users can see links. Click Here To Register...] (Andrea) wrote:


I believe that 8M urea has been used to denature proteins. Denaturing
will almost always deactivate an enzyme.

Steve Turner

Real address contains worldnet instead of spamnet

William Penrose 11-14-2003 12:50 AM

[x-post] Enzyme deactivation
 
On Thu, 13 Nov 2003 23:31:16 GMT, Steve Turner
<[Only registered and activated users can see links. Click Here To Register...].net> wrote:


Some proteins will regenerate activity when the urea is removed, which
by the way you would probably have to do. 8M urea is pretty thick.

Some people use diethylcarbonate to destroy enzymes. It has to be
handled with extreme care, but it decomposes into ethanol and CO2
after it's done its thing. Be sure to study it carefully before even
buying it, since it can decompose and blow up in the refrigerator if
not stored. I don't know its toxic properties.

Bill Penrose


Andrea 11-14-2003 07:26 AM

[x-post] Enzyme deactivation
 
William Penrose <[Only registered and activated users can see links. Click Here To Register...]> wrote in message news:<[Only registered and activated users can see links. Click Here To Register...]>. ..

Mmmh...I'll get sure about toxic properties, but the idea sounds good enough!


Wyrin 11-14-2003 04:06 PM

[x-post] Enzyme deactivation
 
"Andrea" <[Only registered and activated users can see links. Click Here To Register...]> wrote in message
news:57d41e00.0311122337.490ce2c4@posting.google.c om...

Depends if you want an easy step and want your product free from impurities

Add an (competitive) inhibitor of the cellulase at high concentration
[Only registered and activated users can see links. Click Here To Register...]
has a list of inhibitors - 2,4-dinitrophenol might be a good bet.
EDTA will strip away metal ions that stimulate activity
10 % SDS claims to work

But all these might require a clean up step afterwards if you need pure
products



AM 11-15-2003 11:57 AM

[x-post] Enzyme deactivation
 

"Wyrin" <[Only registered and activated users can see links. Click Here To Register...]> wrote:>
impurities

I understand, but I don't need a high purity product.
However I'm going to subscribe Brenda....it seems to be a good resource.
Bye



EK 11-21-2003 04:17 PM

[x-post] Enzyme deactivation
 
Andrea,
Check Brenda ensyme database [Only registered and activated users can see links. Click Here To Register...]
I would try glucose, some metals (Hg2+, Ni2+), or detergents (SDS, Teepol
....)
Emir

"Andrea" <[Only registered and activated users can see links. Click Here To Register...]> wrote in message
news:57d41e00.0311122337.490ce2c4@posting.google.c om...



AM 11-21-2003 10:26 PM

[x-post] Enzyme deactivation
 

"EK" <[Only registered and activated users can see links. Click Here To Register...]> ha scritto nel messaggio
news:zUqvb.27$[Only registered and activated users can see links. Click Here To Register...]...

Thanks, I subcribed Brenda some days ago and I found it very useful.
I think I'll try with SDS and some other, just to begin ( everyone but Hg or
I'll have some troubles with the local environment agency....;| )
However, I'd like to thank you all for your kindness.
See you soon,

Andrea



EK 11-22-2003 05:45 AM

[x-post] Enzyme deactivation
 

"AM" <[Only registered and activated users can see links. Click Here To Register...]> wrote in message
news:liwvb.2736$[Only registered and activated users can see links. Click Here To Register...].it...
Teepol
or

Always happy to help, Andrea. Good luck in your endeavor! I recommend you to
crosspost your questions to other newsgroups, e.g. (for this specific case)
bionet.molbio.methds-reagnts or bionet.proteins in the future.
Cheers,
Emir



John Savage 11-23-2003 11:26 PM

[x-post] Enzyme deactivation
 
[Only registered and activated users can see links. Click Here To Register...] (Andrea) writes:

I'm no chemist, but I do know that Chinese cooks destroy the enzyme in
pineapple using strong saline. Have you tried salty water?
--
John Savage (news address invalid; keep news replies in newsgroup)



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