Ancestral Sequence Reconstruction Enhances Gene Mining Efforts for Industrial Ene Reductases by Expanding Enzyme Panels with Thermostable Catalysts,ACS Catalysis

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Ancestral Sequence Reconstruction Enhances Gene Mining Efforts for Industrial Ene Reductases by Expanding Enzyme Panels with Thermostable Catalysts
ACS Catalysis ( IF 11.3 ) Pub Date : 2023-02-06 , DOI: 10.1021/acscatal.2c03859
Jovan Livada ₁ , Ariana M. Vargas ₁ , Carlos A. Martinez ₁ , Russell D. Lewis ₁

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Ancestral sequence reconstruction (ASR) has been used to produce enzymes that exhibit enhanced stability, activity, and promiscuity compared to wild-type (WT) enzymes. In this work, we compiled a panel of 57 closely related wild-type ene reductases and 56 counterpart ancestral enzymes and statistically compared their biocatalytic properties. Enzyme activity, selectivity, thermostability, and expression level distributions were used to compare their performance. ASR and WT enzymes exhibited comparable activity and overexpression levels, but ASR enzymes displayed enhanced average thermostability (by 9 °C) compared to their wild-type counterparts. Our results demonstrate that ASR-derived ene reductase enzymes may be used as a complement to wild-type enzymes and may provide a superior source of biocatalysts for alkene reduction for industrial applications.

中文翻译：

祖先序列重建通过使用耐热催化剂扩展酶组来增强工业烯还原酶的基因挖掘工作

祖先序列重建 (ASR) 已用于生产与野生型 (WT) 酶相比表现出增强的稳定性、活性和混杂性的酶。在这项工作中，我们编制了一组 57 种密切相关的野生型烯还原酶和 56 种对应的祖先酶，并统计比较了它们的生物催化特性。酶活性、选择性、热稳定性和表达水平分布被用来比较它们的性能。ASR 和 WT 酶表现出相当的活性和过表达水平，但与野生型对应物相比，ASR 酶表现出增强的平均热稳定性（9 °C）。

更新日期：2023-02-06

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