Enantioselective synthesis of (S)-phenylephrine by recombinant Escherichia coli cells expressing the short-chain dehydrogenase/reductase gene from Serratia quinivorans BCRC 14811
| Autor: | Ming-Te Yang, Wen-Hwei Hsu, Yen-Ching Cho, Guan-Jhih Peng, Tze-Kai Fu |
|---|---|
| Rok vydání: | 2013 |
| Předmět: |
chemistry.chemical_classification
Short-chain dehydrogenase biology Stereochemistry Bioengineering Dehydrogenase Reductase medicine.disease_cause Applied Microbiology and Biotechnology Biochemistry Cofactor Enzyme chemistry biology.protein medicine bacteria Enantiomeric excess Escherichia coli Alcohol dehydrogenase |
| Zdroj: | Process Biochemistry. 48:1509-1515 |
| ISSN: | 1359-5113 |
| Popis: | 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 ± 2.3 U/mg protein and 0.24 ± 0.01 U/mg protein, respectively, at 30 °C 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. |
| Databáze: | OpenAIRE |
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