Stereoselective synthesis of (R)-phenylephrine using recombinant Escherichia coli cells expressing a novel short-chain dehydrogenase/reductase gene from Serratia marcescens BCRC 10948

• Published in: Journal of biotechnology 2014-01, Vol.170, p.6-9
• Main Authors: Peng, Guan-Jhih, Kuan, Yi-Chia, Chou, Hsiao-Yi, Fu, Tze-Kai, Lin, Jia-Shin, Hsu, Wen-Hwei, Yang, Ming-Te
• Format: Article
• Language: English
• Subjects: (R)-Phenylephrine; Bacterial Proteins - genetics; Bacterial Proteins - metabolism; Biocatalyst; Enantioselective synthesis; Escherichia coli - genetics; Escherichia coli - metabolism; Fructose - metabolism; NAD - metabolism; NADP - metabolism; Oxidoreductases - genetics; Oxidoreductases - metabolism; Phenylephrine - analogs & derivatives; Phenylephrine - metabolism; Phylogeny; Recombinant Proteins - genetics; Recombinant Proteins - metabolism; Serratia marcescens; Serratia marcescens - enzymology; Serratia marcescens - genetics; Short-chain dehydrogenase/reductase; Substrate Specificity
• ISSN: 0168-1656; 1873-4863
• DOI: 10.1016/j.jbiotec.2013.11.011

•A short-chain dehydrogenase/reductase (SM_SDR) from Serratia marcescens.•SM_SDR prefers NADPH as cofactor.•E. coli BL21 (DE3) expressing SM_SDR can produce (R)-PE from HPMAE.•Fructose is the most effective carbon source for biocatalysis. (R)-Phenylephrine [(R)-PE] is an α1-adrenergic receptor agonist and is widely used as a nasal decongestant to treat the common cold without the side effects of other ephedrine adrenergic drugs. We identified a short-chain dehydrogenase/reductase (SM_SDR) from Serratia marcescens BCRC 10948 that was able to convert 1-(3-hydroxyphenyl)-2-(methylamino) ethanone (HPMAE) into (R)-PE. The SM_SDR used NADPH and NADH as cofactors with specific activities of 17.35±0.71 and 5.57±0.07mU/mg protein, respectively, at 30°C and pH 7.0, thereby indicating that this enzyme could be categorized as an NADPH-preferring short-chain dehydrogenase/reductase. Escherichia coli strain BL21 (DE3) expressing SM_SDR could convert HPMAE into (R)-PE with more than 99% enantiomeric excess. The productivity and conversion yield were 0.57mmolPE/lh and 51.06%, respectively, using 10mM HPMAE. Fructose was the most effective carbon source for the conversion of HPMAE to (R)-PE.