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Directed Evolution of a Hydroxylase into a Decarboxylase for Synthesis of 1-Alkenes from Fatty Acids
ACS Catalysis ( IF 11.3 ) Pub Date : 2020-11-24 , DOI: 10.1021/acscatal.0c04345 Shuaibo Wang 1 , Shengsheng Jiang 1 , Hao Chen 1 , Wen-Ju Bai 2 , Xiqing Wang 1
ACS Catalysis ( IF 11.3 ) Pub Date : 2020-11-24 , DOI: 10.1021/acscatal.0c04345 Shuaibo Wang 1 , Shengsheng Jiang 1 , Hao Chen 1 , Wen-Ju Bai 2 , Xiqing Wang 1
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Inspired by the catalytic mechanism of cytochrome P450 enzymes, we have evolved a fatty acid hydroxylase, P450BSβ, into a decarboxylase after four rounds of saturation mutagenesis that now predominantly converts fatty acids to 1-alkenes. The engineered enzyme directly utilizes inexpensive H2O2 to oxidatively decarboxylate C6–C18 saturated fatty acids as well as plant oil-derived unsaturated fatty acids, displaying high reactivity (e.g., 3,700 h–1 of turnover frequency for myristic acid). With in situ H2O2 supplied by glucose oxidase, this enzyme delivers 1-alkenes in a more productive and scalable fashion (up to 99% yield). Our directed evolution efforts present a strategy for developing new fatty acid decarboxylase for downstream biocatalytic applications.
中文翻译：
定向进化为从脂肪酸合成1-烯烃的羟化酶到脱羧酶
受细胞色素P450酶催化机制的启发，在经过四轮饱和诱变后，我们已经将脂肪酸羟化酶P450BSβ演化为脱羧酶，现在主要将脂肪酸转化为1-烯烃。该工程酶直接利用廉价的H 2 O 2将C6-C18饱和脂肪酸以及植物油衍生的不饱和脂肪酸氧化脱羧，表现出很高的反应性（例如，肉豆蔻酸的周转频率为3700 h -1）。带原位H 2 O 2由葡萄糖氧化酶提供，该酶以更高的产量和可扩展的方式（高达99％的产率）提供1-烯烃。我们的定向进化努力提出了开发用于下游生物催化应用的新型脂肪酸脱羧酶的策略。
更新日期：2020-12-18
中文翻译：
定向进化为从脂肪酸合成1-烯烃的羟化酶到脱羧酶
受细胞色素P450酶催化机制的启发，在经过四轮饱和诱变后，我们已经将脂肪酸羟化酶P450BSβ演化为脱羧酶，现在主要将脂肪酸转化为1-烯烃。该工程酶直接利用廉价的H 2 O 2将C6-C18饱和脂肪酸以及植物油衍生的不饱和脂肪酸氧化脱羧，表现出很高的反应性（例如，肉豆蔻酸的周转频率为3700 h -1）。带原位H 2 O 2由葡萄糖氧化酶提供，该酶以更高的产量和可扩展的方式（高达99％的产率）提供1-烯烃。我们的定向进化努力提出了开发用于下游生物催化应用的新型脂肪酸脱羧酶的策略。
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