A Novel NADH-Dependent Carbonyl Reductase fromKluyveromyces aestuariiand Comparison of NADH-Regeneration System for the Synthesis of Ethyl (S)-4-Chloro-3-hydroxybutanoate

A novel carbonyl reductase from Pichia stipitis for the production of ethyl (S)-4-chloro-3-hydroxybutanoate

“…The SDRs form a well-established enzyme family of oxidoreductases distinct from functionally related enzyme families such as aldoketo reductases that have previously been shown to reduce COBE to (R)-CHBE (Kita et al 1996). Several reductases in SDRs have catalyzed the production of (S)-CHBE with greater than 99% e.e., such as S1 from Candida magnoliae (Yasohara et al 2000) and KaCR from Kluyveromyces aestuarii (Yamamoto et al 2004). Amino acid sequence alignment showed only 59.1% identity between PsCR and S1, and 42.2% identity between PsCR and KaCR.…”

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

“…Enzyme activities were examined using COBE as the substrate. The recombinant PsCR II exhibited an activity of 15 U/mg protein with NADH as an electron donor, which was about 3-fold higher than the activity of another carbonyl reductase, E. coli HB101 harboring pSE-KaCR, which also uses NADH for catalysis (Yamamoto et al, 2004). The activity of E. coli with pET-22b-PsCR II was the highest ever reported for reductases producing (S)-CHBE, with the exception of PsCR I, which required NADPH as an electron donor in our previous studies (Ye et al, 2009b).…”

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

“…The reaction was monitored at 30°C by absorbance at 340 nm. The enzyme activity was determined spectrophotometrically as described previously (Yamamoto et al, 2004;Ye et al, 2009). One unit of reductase activity was defined as the amount catalyzing the reduction of 1 lmol of NADH per minute.…”