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Natural Science Forum / Biology / Biology / November 2003[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 >Hi everybody, here is the question: > [quoted text clipped - 13 lines] > >Andrea Mitarotonda 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 >>I tried to deactivate the enzyme by high temperature, but my product >>doesn't like very much heat. > >I believe that 8M urea has been used to denature proteins. Denaturing >will almost always deactivate an enzyme. 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 > >>I tried to deactivate the enzyme by high temperature, but my product > >>doesn't like very much heat. [quoted text clipped - 10 lines] > buying it, since it can decompose and blow up in the refrigerator if > not stored. I don't know its toxic properties. Mmmh...I'll get sure about toxic properties, but the idea sounds good enough! > Bill Penrose > Hi everybody, here is the question: > [quoted text clipped - 8 lines] > Do you know other ways to deactivate the enzyme. > Have you any advice for me? Depends if you want an easy step and want your product free from impurities Add an (competitive) inhibitor of the cellulase at high concentration http://www.brenda.uni-koeln.de/php/result_flat.php3?ecno=3.2.1.4 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" <dsy21@hotmail.com> wrote:> > Depends if you want an easy step and want your product free from impurities > [quoted text clipped - 6 lines] > But all these might require a clean up step afterwards if you need pure > products 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 http://www.brenda.uni-koeln.de I would try glucose, some metals (Hg2+, Ni2+), or detergents (SDS, Teepol ...) Emir > Hi everybody, here is the question: > [quoted text clipped - 13 lines] > > Andrea Mitarotonda > Andrea, > Check Brenda ensyme database http://www.brenda.uni-koeln.de > I would try glucose, some metals (Hg2+, Ni2+), or detergents (SDS, Teepol > ...) > Emir 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 > > Andrea, > > Check Brenda ensyme database http://www.brenda.uni-koeln.de [quoted text clipped - 9 lines] > > Andrea 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 >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'm no chemist, but I do know that Chinese cooks destroy the enzyme in pineapple using strong saline. Have you tried salty water?  SignatureJohn Savage (news address invalid; keep news replies in newsgroup) > >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'm no chemist, but I do know that Chinese cooks destroy the enzyme in > pineapple using strong saline. Have you tried salty water? I can't use high salt concentration because of the product I treat. But I have no doubts that chinese cooking can destroy enzymes...!!! >> I'm no chemist, but I do know that Chinese cooks destroy the enzyme in >> pineapple using strong saline. Have you tried salty water? > >>I can't use high salt concentration because of the product I treat. >>But I have no doubts that chinese cooking can destroy enzymes...!!! I was referring to treating raw pineapple, actually. SignatureJohn Savage (news address invalid; keep news replies in newsgroup) |
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