Enantioselective synthesis of (S)-phenylephrine by whole cells of recombinant Escherichia coli expressing the amino alcohol dehydrogenase gene from Rhodococcus erythropolis BCRC 10909

Stereoselective synthesis of (R)-phenylephrine using recombinant Escherichia coli cells expressing a novel short-chain dehydrogenase/reductase gene from Serratia marcescens BCRC 10948

“…Recently, we found a short-chain dehydrogenase/reductase gene (SQ SDR) from Serratia quinivorans BCRC 14811, with high catalytic activity and an 80-fold preference for NADH over NADPH as a cofactor (Peng et al, 2013). E. coli cells expressing NADH-dependent SQ SDR exhibited a much higher conversion yield and productivity for the conversion of HPMAE to (S)-PE than our previous strain, E. coli expressing RE AADH (Lin et al, 2010;Peng et al, 2013). Although (S)-PE can be produced effectively by recombinant E. coli cells, (S)-PE must be converted into (R)-PE by Walden inversion reaction for medical use.…”

“…The cloned gene (designated SM SDR) in pET30a-sdr10 contained a 750-bp ORF and showed 98% identity to the gene sequence of a putative oxidoreductase (GenBank: AGE20192) from S. marcescens WW4. The calculated molecular mass of SM SDR is 27.4 kDa, and the amino acid sequence of SM SDR showed 33.7% identity to SQ SDR from S. quinivorans BCRC 14811 (Peng et al, 2013) and 30.2% identity to RE AADH from R. erythropolis BCRC 10909 (Lin et al, 2010).…”

“…We developed a biotransformation process for limited production of (S)-PE from 1-(3-hydroxyphenyl)-2-(methylamino) ethanone (HPMAE) using recombinant Escherichia coli expressing an NADPH-dependent amino alcohol dehydrogenase gene (RE AADH) from Rhodococcus erythropolis BCRC 10909 (Lin et al, 2010). Recently, we found a short-chain dehydrogenase/reductase gene (SQ SDR) from Serratia quinivorans BCRC 14811, with high catalytic activity and an 80-fold preference for NADH over NADPH as a cofactor (Peng et al, 2013).…”

“…In a cell-free process using isolated RE AADH as biocatalyst for the production of (S)-PE from HPMAE, NADPH is required as a cofactor for the reduction activity of RE AADH (Lin et al, 2010). Because NADPH is an unstable molecule and is very expensive, E. coli cells expressing SM SDR were used as biocatalysts to convert HPMAE into (R)-PE and to efficiently replenish the NADPH in the biocatalysis process.…”

“…4) revealed that SM SDR, which is capable of converting HPMAE into (R)-PE, was in a separated branch from SQ SDR and RE AADH, which can convert HPMAE into (S)-PE (Lin et al, 2010;Peng et al, 2013). RE AADH and SQ SDR were in the same cluster, suggesting that the common ancestor of these enzymes has the ability to reduce HPMAE to (S)-PE.…”