Enantioselective synthesis of (S)-phenylephrine by whole cells of recombinant Escherichia coli expressing the amino alcohol dehydrogenase gene from Rhodococcus erythropolis BCRC 10909
| Autor: | Wen-Hwei Hsu, Chien-Yu Chen, Huei-Chung Chen, Wei-De Lin |
|---|---|
| Rok vydání: | 2010 |
| Předmět: |
chemistry.chemical_classification
Molecular mass biology Chemistry Bioengineering Dehydrogenase Reductase medicine.disease_cause Applied Microbiology and Biotechnology Biochemistry law.invention Enzyme law medicine biology.protein Enantiomeric excess Walden inversion Escherichia coli Alcohol dehydrogenase |
| Zdroj: | Process Biochemistry. 45:1529-1536 |
| ISSN: | 1359-5113 |
| Popis: | (R)-phenylephrine [(R)-PE] is an α1-adrenergic receptor agonist that is widely used in over-the-counter drugs to treat the common cold. We found that Rhodococcus erythropolis BCRC 10909 can convert detectable level of 1-(3-hydroxyphenyl)-2-(methylamino) ethanone (HPMAE) to (S)-PE by high performance liquid chromatography tandem mass spectrometry analysis. An amino alcohol dehydrogenase gene (RE_AADH) which possesses the ability to convert HPMAE to (S)-PE was then isolated from R. erythropolis BCRC 10909 and expressed in Escherichia coli NovaBlue. The purified RE_AADH, tagged with 6×His, had a molecular mass of approximately 30 kDa and exhibited a specific activity of 0.19 μU/mg to HPMAE in the presence of NADPH, indicating this enzyme could be categorized as NADP+-dependent short-chain dehydrogenase reductase. E. coli NovaBlue cell expressing the RE_AADH gene was able to convert HPMAE to (S)-PE with more than 99% enantiomeric excess (ee), 78% yield and a productivity of 3.9 mmol (S)-PE/L h in 12 h at 30 °C and pH 7. The (S)-PE, recovered from reaction mixture by precipitation at pH 11.3, could be converted to (R)-PE (ee > 99%) by Walden inversion reaction. This is the first reported biocatalytic process for the production of (S)-PE from HPMAE. |
| Databáze: | OpenAIRE |
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