1. Stereoselective synthesis of (R)-phenylephrine using recombinant Escherichia coli cells expressing a novel short-chain dehydrogenase/reductase gene from Serratia marcescens BCRC 10948.
- Author
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Peng GJ, Kuan YC, Chou HY, Fu TK, Lin JS, Hsu WH, and Yang MT
- Subjects
- Bacterial Proteins genetics, Bacterial Proteins metabolism, Escherichia coli genetics, Fructose metabolism, NAD metabolism, NADP metabolism, Oxidoreductases genetics, Phylogeny, Recombinant Proteins genetics, Recombinant Proteins metabolism, Serratia marcescens enzymology, Substrate Specificity, Escherichia coli metabolism, Oxidoreductases metabolism, Phenylephrine analogs & derivatives, Phenylephrine metabolism, Serratia marcescens genetics
- 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., (Copyright © 2013 Elsevier B.V. All rights reserved.)
- Published
- 2014
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