Directed evolution of Aspergillus oryzae lipase for the efficient resolution of (R,S)-ethyl-2-(4-hydroxyphenoxy) propanoate
Directed evolution of Aspergillus oryzae lipase for the efficient resolution of (R,S)-ethyl-2-(4-hydroxyphenoxy) propanoate
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米曲霉脂肪酶的定向进化可有效拆分 (R,S)-乙基-2-(4-羟基苯氧基) 丙酸酯
DOI：
10.1007/s00449-020-02393-7
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发表时间：
2020-09
影响因子：
3.8
通讯作者：
Zheng Jianyong
中科院分区：
文献类型：
--
作者：
Zhang Mengjie;Li Qi;Lan Xing;Li Xiaojun;Zhang Yinjun;Wang Zhao;Zheng Jianyong
米曲霉脂肪酶（AOL）是一种潜在的工业应用生物催化剂。本研究通过两轮定向进化筛选出突变脂肪酶AOL-3F38N/V230R，使脂肪酶活性提高四倍，催化效率（kcat/Km）提高三倍，同时保持其优异的立体选择性。 AOL-3F38N/V230R 酶活性在 pH 7.5 和 40 °C 时达到最大。与野生型AOL-3相比，AOL-3F38N/V230R优先水解脂肪酸乙酯碳链长度从C4到C6–C10。在相同的催化反应条件下，AOL-3F38N/V230R对(R,S)-乙基-2-(4-羟基苯氧基)丙酸酯((R,S)-EHPP)的转化率比原酶提高了169.7%。 (R,S)-EHPP的e.s达到99.4%，转化率约为50.2%，E值为829.0。因此，AOL-3F38N/V230R是获得芳氧基苯氧基丙酸酯（APP）除草剂关键手性化合物的潜在生物催化剂。
Aspergillus oryzae lipase (AOL) is a potential biocatalyst for industrial application. In this study, a mutant lipase AOL-3F38N/V230R was screened through two rounds of directed evolution, resulting in a fourfold increase in lipase activity, and threefold in catalytic efficiency (kcat/Km), while maintaining its excellent stereoselectivity. AOL-3F38N/V230R enzyme activity was maximum at pH 7.5 and also at 40 °C. And compared with wild-type AOL-3, AOL-3F38N/V230R preferentially hydrolyzed the fatty acid ethyl ester carbon chain length from C4 to C6–C10. In the same catalytic reaction conditions, the conversion of (R,S)-ethyl-2-(4-hydroxyphenoxy) propanoate ((R,S)-EHPP) by AOL-3F38N/V230R can be increased 169.7% compared to the original enzyme. The e.e.s of (R,S)-EHPP achieved 99.4% and conversion about 50.2% with E value being 829.0. Therefore, AOL-3F38N/V230R was a potential biocatalyst for obtaining key chiral compounds for aryloxyphenoxy propionate (APP) herbicides.
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影响因子：
1.9
作者：
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通讯作者：
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影响因子：
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作者：
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通讯作者：
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