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Study on the relationship between structure and enantioselectivity of a hyperthermophilic esterase from archaeon Aeropyrum pernix K1 |
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| Authors: | Guirong Zhang Renjun Gao Liangyu Zheng Aijun Zhang Yuanhong Wang Qiuyan Wang Yan Feng Shugui Cao |
| Institution: | aKey Laboratory for Molecular Enzymology and Engineering of Ministry of Education, Jilin University, Changchun 130023, PR China bChangchun Institute of Education, Changchun 130061, PR China cNortheast Normal University, Changchun 130021, PR China |
| Abstract: | To enhance the enantioselectivity of a hyperthermophilic esterase from archaeon Aeropyrum pernix K1 (APE1547), a directed evolution approach is employed to generate mutant library from the native enzyme. A mutation (TBC26) is identified after one round of epPCR. The enantioselectivity of TBC26 is increased up to 2.6-fold compared to that of wild type enzyme. TBC26 contains five amino acid substitutions (R11G, L36P, V223A, I551L, A564T). The five mutation sites are spatially distant to the catalytic center. According to the published crystal structure of WT and considering the changes of secondary and tertiary structure, here we try to explain the change of enantioselectivity of the TBC26. The results suggest that the change of enantioselectivity of enzyme has a close relationship to the configuration of the enzyme. |
| Keywords: | Directed evolution Enantioselectivity Screening 2-Octanol acetate Configuration Mutant |
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