[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

[Only registered users see links. ] (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

On Thu, 13 Nov 2003 23:31:16 GMT, Steve Turner
<[Only registered users see links. ].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

William Penrose <[Only registered users see links. ]> wrote in message news:<[Only registered users see links. ]>. ..

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

"Andrea" <[Only registered users see links. ]> 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 users see links. ]
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

"Wyrin" <[Only registered users see links. ]> 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

Andrea,
Check Brenda ensyme database [Only registered users see links. ]
I would try glucose, some metals (Hg2+, Ni2+), or detergents (SDS, Teepol
....)
Emir

"Andrea" <[Only registered users see links. ]> wrote in message
news:57d41e00.0311122337.490ce2c4@posting.google.c om...

"EK" <[Only registered users see links. ]> ha scritto nel messaggio
news:zUqvb.27$[Only registered users see links. ]...

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

"AM" <[Only registered users see links. ]> wrote in message
news:liwvb.2736$[Only registered users see links. ].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

[Only registered users see links. ] (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)