Stereoselective synthesis of (R)-phenylephrine using recombinant Escherichia coli cells expressing a novel short-chain dehydrogenase/reductase gene from Serratia marcescens BCRC 10948
详细信息 查看全文
- 作者：Guan-Jhih Peng ; Yi-Chia Kuan ; Hsiao-Yi Chou ; Tze-Kai Fu ; Jia-Shin Lin ; Wen-Hwei Hsu ; Ming-Te Yang
- 关键词：ADH ; alcohol dehydrogenase ; AKR ; aldo-keto reductase ; DDBDH ; 2 ; 3-dihydro-2 ; 3-dihydroxybenzoate dehydrogenase ; DHMPR ; dihydromonapterin reductase ; Dro_ADH ; D. melanogaster alcohol dehydrogenase ; GDH ; glucose 1-dehydrogenase B ; HBDH ; d-beta-hydroxybutyrate dehydrogenase ; HPLC ; high performance liquid chromatography ; HPMAE ; 1-(3-hydroxyphenyl)-2-(methylamino) ethanone ; IPTG ; isopropyl-尾-d-thiogalactopyranoside ; KACPR ; 3-ketoacyl-(acyl-carrier-protein) reductase ; KDGR ; 5-keto-d-gluconate-5-reductas
- 刊名：Journal of Biotechnology
- 出版年：20 January, 2014
- 年：2014
- 卷：170
- 期：Complete
- 页码：6-9
- 全文大小：624 K
文摘
(R)-Phenylephrine [(R)-PE] is an 伪1-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 卤 0.71 and 5.57 卤 0.07 mU/mg protein, respectively, at 30 掳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 0.57 mmol PE/l h and 51.06%, respectively, using 10 mM HPMAE. Fructose was the most effective carbon source for the conversion of HPMAE to (R)-PE.