One-pot synthesis of (R)- and (S)-phenylglycinol from bio-based l-phenylalanine by an artificial biocatalytic cascade

Abstract

Chiral phenylglycinol is a very important chemical in the pharmaceutical manufacturing. Current methods for synthesis of chiral phenylglycinol often suffered from unsatisfied selectivity, low product yield and using the non-renewable resourced substrates, then the synthesis of chiral phenylglycinol remain a grand challenge. Design and construction of synthetic microbial consortia is a promising strategy to convert bio-based materials into high value-added chiral compounds. In this study, we reported a six-step artificial cascade biocatalysis system for conversion of bio-based l-phenylalanine into chiral phenylglycinol. This system was designed using a microbial consortium including two engineered recombinant Escherichia coli cell modules, one recombinant E. coli cell module co-expressed six different enzymes (phenylalanine ammonia lyase/ferulic acid decarboxylase/phenylacrylic acid decarboxylase/styrene monooxygenase/epoxide hydrolase/alcohol dehydrogenase) for efficient conversion of l-phenylalanine into 2-hydroxyacetophenone. The second recombinant E. coli cell module expressed an (R)-ω-transaminase or co-expressed the (S)-ω-transaminase, alanine dehydrogenase and glucose dehydrogenase for conversion of 2-hydroxyacetophenone into (S)- or (R)-phenylglycinol, respectively. Combining the two engineered E. coli cell modules, after the optimization of bioconversion conditions (including pH, temperature, glucose concentration, amine donor concentration and cell ratio), l-phenylalanine could be easily converted into (R)-phenylglycinol and (S)-phenylglycinol with up to 99% conversion and > 99% ee. Preparative scale biotransformation was also conducted on 100-mL scale, (S)-phenylglycinol and (R)-phenylglycinol could be obtained in 71.0% and 80.5% yields, > 99% ee, and 5.19 g/L d and 4.42 g/L d productivity, respectively. The salient features of this biocatalytic cascade system are good yields, excellent ee, mild reaction condition and no need for additional cofactor (NADH/NAD+), provide a practical biocatalytic method for sustainable synthesis of (S)-phenylglycinol and (R)-phenylglycinol from bio-based L-phenylalanine. [Figure not available: see fulltext.].