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Efficient protein expression in a robust Escherichia coli strain and its application for kinetic resolution of racemic glycidyl o-methylphenyl ether in high concentration

Biochemical Engineering Journal. 2020-06; 
Xiaoyang Oua , Fei Penga , Xiaoling Wua , Pei Xua , Minhua Zonga,b , Wenyong Loua
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Codon Optimization The codon-optimized genes of epoxide hydrolase (EH, Genbank: KX146840.1), lipase 3646 (Lip3646, Genbank: JX833623), and carbonyl reductase (AcCR, Genbank: MF419650) from Sphingomonas sp. HXN-200, Cohnella sp. A01, and Acetobacter sp. CCTCC M209061, respectively, were synthetized by GenScript Biotech Corp. Get A Quote

摘要

A robust?Escherichia coli?(E. coli) strain, with a dual protection system to fight against microbial contamination and T7 phage infection, is used as a chassis to overexpress various proteins in auxotrophic MOPS medium. Among them, a robust EH (epoxide hydrolase)-producing?E. coli?was used as a biocatalyst to enantioselectively resolve racemic glycidyl?o-methylphenyl ether (rac-o-GMPE) for preparing (R)-o-GMPE with 6416.7 U/g wet cells. The optimal temperature, pH and biocatalyst dosage were 25 °C, 7.0 and 0.2 mg/mL in aqueous buffer, respectively. The maximal yield (23.0 %) and?ee?(99.2 %) were achieved within 12 min in 20 mM?rac-o-GMPE. Additionally, 98.0 %?ee?and 38.7 % yield were observed by cell... More

關鍵詞

Epoxide hydrolase Kinetic resolution (R)-o-GMPE Robust E. coli Biphasic system
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