title: Stereoselective synthesis of (R)-phenylephrine using recombinant Escherichia coli cells expressing a novel short-chain dehydrogenase/reductase gene from Serratia marcescens BCRC 10948.
abstract: (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.
authors.affiliation: Institute of Molecular Biology, National Chung Hsing University, Taichung 402, Taiwan.
authors.forename: Guan-Jhih
authors.initials: GJ
authors.lastname: Peng
authors.affiliation: Institute of Molecular and Cellular Biology & Department of Life Science, National Tsing Hua University, Hsinchu 300, Taiwan.
authors.forename: Yi-Chia
authors.initials: YC
authors.lastname: Kuan
authors.affiliation: Institute of Molecular Biology, National Chung Hsing University, Taichung 402, Taiwan.
authors.forename: Hsiao-Yi
authors.initials: HY
authors.lastname: Chou
authors.affiliation: Institute of Molecular Biology, National Chung Hsing University, Taichung 402, Taiwan.
authors.forename: Tze-Kai
authors.initials: TK
authors.lastname: Fu
authors.affiliation: Institute of Molecular Biology, National Chung Hsing University, Taichung 402, Taiwan.
authors.forename: Jia-Shin
authors.initials: JS
authors.lastname: Lin
authors.affiliation: Institute of Molecular Biology, National Chung Hsing University, Taichung 402, Taiwan. Electronic address: whhsu@dragon.nchu.edu.tw.
authors.forename: Wen-Hwei
authors.initials: WH
authors.lastname: Hsu
authors.affiliation: Institute of Molecular Biology, National Chung Hsing University, Taichung 402, Taiwan. Electronic address: mtyang@dragon.nchu.edu.tw.
authors.forename: Ming-Te
authors.initials: MT
authors.lastname: Yang
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D005632:Fructose / Q000378:metabolism; D009243:NAD / Q000378:metabolism; D009249:NADP / Q000378:metabolism; D010088:Oxidoreductases / Q000235:genetics / Q000378:metabolism*; D010656:Phenylephrine / Q000031:analogs & derivatives* / Q000378:metabolism*; D010802:Phylogeny; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D012706:Serratia marcescens / Q000201:enzymology / Q000235:genetics*; D013379:Substrate Specificity
chemical_list: D001426:Bacterial Proteins; D011994:Recombinant Proteins; D009243:NAD; D010656:Phenylephrine; D005632:Fructose; D009249:NADP; D010088:Oxidoreductases
keywords: (R)-Phenylephrine; 1-(3-hydroxyphenyl)-2-(methylamino) ethanone; 2,3-dihydro-2,3-dihydroxybenzoate dehydrogenase; 3-ketoacyl-(acyl-carrier-protein) reductase; 3-oxoacyl-(acyl-carrier-protein) reductase; 5-keto-d-gluconate-5-reductase; ADH; AKR; Biocatalyst; D. melanogaster alcohol dehydrogenase; DDBDH; DHMPR; Dro_ADH; Enantioselective synthesis; GDH; HBDH; HPLC; HPMAE; IPTG; KACPR; KDGR; L. brevis R-alcohol dehydrogenase; LB_RADH; OACPR; PE; POACPR; POR; PSOR; R-alcohol dehydrogenase; R. erythropolis amino alcohol dehydrogenase; RADH; RDH; RE_AADH; S. marcescens short-chain dehydrogenase/reductase; S. quinivorans short-chain dehydrogenase/reductase; SDH; SDR; SM_SDR; SPDH; SQ_SDR; Serratia marcescens; Short-chain dehydrogenase/reductase; Z. mobilis ribitol dehydrogenase; ZM_RDH; alcohol dehydrogenase; aldo-keto reductase; d-beta-hydroxybutyrate dehydrogenase; dihydromonapterin reductase; glucose 1-dehydrogenase B; high performance liquid chromatography; isopropyl-β-d-thiogalactopyranoside; phenylephrine; putative NAD(P)-binding oxidoreductase; putative oxoacyl-(acyl-carrier-protein) reductase; putative short-chain oxidoreductase; ribitol dehydrogenase; short-chain dehydrogenase; short-chain dehydrogenase/reductase; sorbitol-6-phosphate dehydrogenase