Biosynthesis of (S)-4-chloro-3-hydroxybutanoate ethyl using Escherichia coli co-expressing a novel NADH-dependent carbonyl reductase and a glucose dehydrogenase

Purification and characterization of a novel NADH-dependent carbonyl reductase from Pichia stipitis involved in biosynthesis of optically pure ethyl (S)-4-chloro-3-hydroxybutanoate

“…The resulting plasmid pET-22b-PsCRII was transformed into E. coli Rosetta (DE3) plysS. Details on cloning and expression have been described previously (Ye et al, 2010c).…”

“…Amino acid sequence analysis grouped PsCR II with the short-chain dehydrogenase/reductase superfamily, with less than 55% identity to known reductases that produce (S)-CHBE. Compared to known reductases PsCR II was more suitable for production because NADH is less expensive than NADPH and the activity of crude enzyme was the highest ever reported for reductases that required NADH for catalysis in the literature to date (Ye et al, 2010c). In this study, recombinant PsCRII was purified and characterized.…”

Improved biosynthesis of ethyl (S)-4-chloro-3-hydroxybutanoate by adding l-glutamine plus glycine instead of NAD+ in β-cyclodextrin–water system

“…In recent years, asymmetric bioreduction of prochiral ketone into highly optically active alcohols has been of great interest due to its mild reaction condition and high stereoselectivity (Ema et al, 2006(Ema et al, , 2008He et al, 2014c;Wang et al, 2012;Ye et al, 2010a). Many reductases have been employed to biotransform COBE to (S)-CHBE with excellent enantioselectivity and high yield (He et al, 2014b;Wang et al, 2011;Ye et al, 2010b).…”

Asymmetric synthesis of (S)-4-chloro-3-hydroxybutanoate by sorbose reductase from Candida albicans with two co-existing recombinant Escherichia coli strains

“…PsCR 5) and PsCRII 6) from Pichia stipitis, S1 from Candida magnoliae, 7) and CpCR from Candida parapsilosis. 8) Glucose dehydrogenase was coupled with all of these reductases for coenzyme regeneration because of its high stability and cheap substrate.…”