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Site-Specific Immobilization of Recombine Esterase APE1547

Author: JuLinZuo
Tutor: GaoRenJun
School: Jilin University
Course: Biochemistry and Molecular Biology
Keywords: APE 1547 site-specific immobilization recombine esterase
CLC: Q814.2
Type: Master's thesis
Year: 2011
Downloads: 23
Quote: 0
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Abstract


Esterase which has been deeply researched in structure and property, is a kind of lipolytic enzyme. It plays an important role in industry. However, the free esterase can’t be reused and hardly recycled. The separation of enzyme with product maybe increase the cost which restrict the development of enzyme. The immobilization of enzyme can solve these problems. Because it is convenience for reusing and recycling and save the cost of production. The immobilized enzyme can be easily separated from product. The optimal temperature of thermophilic esterase is higher than other enzymes, so we can increase the temperature to remove the contamination. It is best to save the energy in process.Site-specific immobilization may protect the naturly structure as much as possible. The specific linkage ensure the volume of enzyme.In this experiment, we used the site-specific method to immobilize recombine thermophilic esterase APE 1547. Two similar carriers were used for immobilization. The substate was p-nitropheno caprylate.The carriers were HZD-2 and HD-2, both of them were the cationic resin. Comparison the different methods could help us to learn the properties of the esterase. We could find the influence of the specific site by recombination.After immobilization, the influence of temperature and pH value decreased. So immobilized enzyme showed higher stability than free enzyme, but lower activity. The recombination had a lower activity than free esterases and big differences in different conditions. Maybe the structures of the protein was the key.We obtain the recombination of hyperthermophilic esterase APE 1547 and immobilization with site-specific method. The problem of unrecyclying and low tolerance were mainly solved. The steric hindrance was at a lower level. Through this experiment, we had a deeper realization for the steric structure of APE 1547. We hope it would be a guidance for the application in industry and our livelihood.

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CLC: > Biological Sciences > Bioengineering ( Biotechnology ) > Enzyme Engineering > Immobilized enzyme and immobilized cell technology
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