Rational design of the carbonyl reductase EbSDR8 for efficient biosynthesis of enantiopure ( R )-3-chloro-1-phenyl-1-pro
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BIOTECHNOLOGICALLY RELEVANT ENZYMES AND PROTEINS
Rational design of the carbonyl reductase EbSDR8 for efficient biosynthesis of enantiopure (R)-3-chloro-1-phenyl-1-propanol Ze-Hui Shao 1
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Bing-Mei Su 2
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Sheng-Li Yang 1
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Li-Dan Ye 3
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Hong-Wei Yu 3
Received: 10 March 2020 / Revised: 1 September 2020 / Accepted: 10 September 2020 # Springer-Verlag GmbH Germany, part of Springer Nature 2020
Abstract (R)-3-Chloro-1-phenyl-1-propanol ((R)-CPPO) is an important chiral intermediate for antidepressants. For its efficient biosynthesis, the carbonyl reductase EbSDR8 was engineered to asymmetrically reduce the unnatural substrate 3-chloro-1-phenyl-1-propanone (3-CPP) at high concentrations. Molecular docking and molecular dynamics simulations of the resulting mutants suggested enlarged substrate binding pocket and more reasonable interactions between the enzyme and the substrate or cofactor as the reasons for the enhanced catalytic activity and thus the remarkably improved conversion of high-concentration 3-CPP. Using the best mutant EbSDR8G94A/L153I/Y188A/Y202M as the whole-cell biocatalyst, reduction of 3-CPP (1.0 M) was conducted using 100% isopropanol as both the solvent and co-substrate for NADH regeneration, delivering (R)-CPPO with ˃ 99% eep and 95.5% conversion. This result suggests EbSDR8G94A/L153I/Y188A/Y202M as a potential biocatalyst for green production of (R)-CPPO at the industrial scale. Key points • Rational design of EbSDR8 by modulating steric hindrance and molecular interactions; • Non-aqueous biocatalysis using isopropanol as both the solvent and co-substrate; • Whole-cell catalyzed production of 161 g/L enantiopure (R)-CPPO from 1.0 M of 3-CPP. Keywords Asymmetric reduction . Steric hindrance . Molecular interactions . Non-aqueous biocatalysis . (R)-3-Chloro-1-phenyl-1-propanol
Introduction Optically pure enantiomers of chiral alcohols are important building blocks for the production of chiral drugs (Huang Electronic supplementary material The online version of this article (https://doi.org/10.1007/s00253-020-10904-5) contains supplementary material, which is available to authorized users. * Sheng-Li Yang [email protected] * Li-Dan Ye [email protected] * Hong-Wei Yu [email protected] 1
College of Pharmaceutical Science, Zhejiang University of Technology, Hangzhou 310014, China
2
College of Chemical Engineering, Fuzhou University, Fuzhou 350116, China
3
Institute of Bioengineering, College of Chemical and Biological Engineering, Zhejiang University, Hangzhou 310027, China
et al. 2015; Liang et al. 2009), fragrances (Homola et al. 2015), agrochemicals (Otoguro et al. 2008), and functional materials (Du et al. 2014). (R)-CPPO is a chiral precursor for antidepressant selective serotonin reuptake inhibitors (SSRIs) such as fluoxetine, nisoxetine, and atomoxetine (Srebnik et al. 1988), and it is also used in the production of the premature ejaculation drug dapoxetine (Dodda et al. 2013). Meanwhile, its isomer (R)-3′-chloro-1-phenyl-1-propanol can be used to synthesize a highly e
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