Green Synthesis and Catalysis
Green Synthesis and Catalysis
4 years ago

Redesign of a short-chain dehydrogenase/reductase for asymmetric synthesis of ethyl (R)-2-hydroxy-4-phenylbutanoate based on per-residue free energy decomposition and sequence conservatism analysis

Redesign of a short-chain dehydrogenase/reductase for asymmetric synthesis of ethyl (<em>R</em>)-2-hydroxy-4-phenylbutanoate based on per-residue free energy decomposition and sequence conservatism analysis
Bingmei Su, Lian Xu, Xinqi Xu, Lichao Wang, Aipeng Li, Juan Lin, Lidan Ye, Hongwei Yu
As an important building block for the synthesis of angiotensin-converting enzyme inhibitors, ethyl (R)-2-hydroxyl-4-phenylbutanoate [(R)-HPBE] has attracted increasing attention. The key to industrial biosynthesis of (R)-HPBE is a biocatalyst that efficiently reduces ethyl 2-oxo-4-phenylbutanoate (OPBE) with high R-enantioselectivity. This paper proposed a strategy for identifying key residues involved in enantioselectivity control based on per-residue free energy decomposition and sequence conservatism analysis. Using this strategy, 4 nonconservative sites with high energy contribution to binding of OPBE were chosen as engineering targets, generating variant Mu27 with 99% conversion and 98% (R) ee value at substrate loading of up to 500 ​mmol/L. MD simulations suggested the higher stability and formation probability of Mu27-OPBEproR prereaction state as key reasons for the excellent R-enantioselectivity of Mu27 towards OPBE. The success in this study provides a viable approach for rational design of alcohol dehydrogenases with high enantioselectivity towards unnatural substrates. Rational design of biocatalyst for efficiently synthesizing (R)-HPBE at engineering targets identified with per-residue free energy decomposition and sequence conservatism analysis.
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Publisher URL: https://www.sciencedirect.com/science/article/pii/S2666554920300302

Open URL: https://doi.org/10.1016/j.gresc.2020.09.003

DOI: 10.1016/j.gresc.2020.09.003

Open access
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