Enantioselective synthesis of (S)-phenylephrine by recombinant Escherichia coli cells expressing the short-chain dehydrogenase/reductase gene from Serratia quinivorans BCRC 14811

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• 作者：Guan-Jhih Peng ; Yen-Ching Cho ; Tze-Kai Fu ; Ming-Te Yang ; Wen-Hwei Hsu
• 关键词：Serratia quinivorans ; Short-chain dehydrogenase/reductase ; Enantioselective synthesis ; Phenylephrine ; Biocatalyst
• 刊名：Process Biochemistry
• 出版年：2013
• 出版时间：October, 2013
• 年：2013
• 卷：48
• 期：10
• 页码：1509-1515
• 全文大小：1807 K

文摘

Background

An amino alcohol dehydrogenase gene (RE_AADH) from Rhodococcus erythropolis BCRC 10909 has been used for the conversion of 1-(3-hydroxyphenyl)-2-(methylamino) ethanone (HPMAE) to (S)-phenylephrine [(S)-PE]. However RE_AADH uses NADPH as cofactor, and only limited production of (S)-PE from HPMAE is achieved.

Methods

A short-chain dehydrogenase/reductase gene (SQ_SDR) from Serratia quinivorans BCRC 14811 was expressed in Escherichia coli BL21 (DE3) for the conversion of HPMAE to (S)-PE.

Results

The SQ_SDR enzyme was capable of converting HPMAE to (S)-PE in the presence of NADH and NADPH, with specific activities of 26.5 ¡À 2.3 U/mg protein and 0.24 ¡À 0.01 U/mg protein, respectively, at 30 ¡ãC and at a pH of 7.0. The E. coli BL21 (DE3), expressing NADH-preferring SQ_SDR, converted HPMAE to (S)-PE with more than 99 % enantiomeric excess, a conversion yield of 86.6 % and a productivity of 20.2 mmol/l h, which was much higher than our previous report using E. coli NovaBlue expressing NADPH-dependent RE_AADH as the biocatalyst.

Conclusion

The SQ_SDR enzyme with its high catalytic activity and strong preference for NADH as a cofactor provided a significant advantage in bioreduction.