I would like to know the best approaches for enzyme immobilization in order to avoid/solve the loss of enzyme activiy during immobilization. I've already try encapsulation but at the end of one week enzyme lost its activity.
Hi Artur, are you losing enzyme activity upon immobilization, or is this a progressive loss of the immobilized enzyme activity with time?
If you are working with immobilized proteinases, then the resin should be stored in the presence of non-covalent inhibitors or excess protein to prevent autolysis.
Hi Artur.I think encapsulation method for enzyme has less efficiency among other methods of immobilization. can substrate move easily from membrane to access enzyme?you can raise your assay time for immobilized enzyme.immobilization time can effect on enzyme structure,decrease immobilization time,another that enzyme conformation change dramaticly when immobilized so activity decrease
Following are 2 papers which discuss on choosing right encapsulation technique.
Chapter Immobilization of Enzymes Acting on Macromolecular Substrate...
Article Parameters Affecting the Performance of Immobilized Enzyme
Dear Steingrimur Stefansson,
i'm loosing my activity along time. At the end of one week i still have the same activity as the free enzyme but at the end of two weeks i lost almost all the activity. Indeed i'm working with a proteinase. Thanks for the sugestion.
Regards
Enzyme is easy to lose activity in soluton, and most immobilization methods were tend to retain part of its activity, and focused on the enzyme reuse ability.
The encapsulation often had poor activity, cause the carrier prevented the enzyme from contacting with the substrate. If you make the carrier in nano or micro size, the problem may be solved.
I am not the sure whether the activity loss happened before or after the immobilization, but If you want to use robust enzyme,I highly recomand cytochrome C.
If you want to find review,make good use of GOOGLE
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