I am going to assess the ability of Cell free extract(taken from potential bacterial cells) to degrade the aromatic substance. So for checking the degradation activity i need to incubate the cell free extract for upto 48hrs in the medium containing aromatic substrate. So, to ensure the enzymes to be in active state which type of buffer would be more preferable.
Engelbert Buxbaum
There are several aspects to your question:
- You have to provide a suitable environment for your enzyme to work (pH, cofactors...). The best source for such info is BRENDA (http://www.brenda-enzymes.info/)
- You have to prevent growth of bacteria and yeasts during such long-term incubations. One possibility is to work aseptically (with sterile-filtered solutions, sterile tools etc.), the other to add preservatives. The latter must not harm the enzyme, again, BRENDA can help.
- The stability of your enzyme has to be high enough to survive the long incubation time. You'll need to check this with suitable control experiments, and you'll have to redo those each time you change something (incubation conditions, mutated enzyme...).
1 votes
0 thanks
Vijaya Sarathy
Dear Engelbert Buxbaum sir,
Thank you a lot for giving me a wonderful suggestion. Yes even i plan to filter the crude cell free extract with 0.45 or 0.24micron filter so that i can make the cells not to enter into the incubating medium..
Yes I need to optimize the conditions to make suit for enzymatic activity...
0 votes
0 thanks
- Badges
- Science topic
- Similar topics
- Phytochemistry
- Extracts
More Vijaya Sarathy's questions See All
Similar questions and discussions