Back to Search
Start Over
Biocatalytic synthesis of ( S)-4-chloro-3-hydroxybutanoate ethyl ester using a recombinant whole-cell catalyst.
- Source :
- Applied Microbiology & Biotechnology; Nov2010, Vol. 88 Issue 6, p1277-1285, 9p
- Publication Year :
- 2010
-
Abstract
- cofactor regeneration system for enzymatic biosynthesis was constructed by coexpressing a carbonyl reductase from Pichia stipitis and a glucose dehydrogenase from Bacillus megaterium in Escherichia coli Rosetta (DE3) PlySs. Transformants containing the polycistronic plasmid pET-PII-SD2-AS1-B exhibited an activity of 13.5 U/mg protein with 4-chloro-3-oxobutanoate ethyl ester (COBE) as the substrate and an activity of 14.4 U/mg protein with glucose as the substrate; NAD(H) was the coenzyme in both cases. Asymmetric reduction of COBE to ( S)-4-chloro-3-hydroxybutanoate ethyl ester [( S)-CHBE] with more than 99% enantiomeric excess was demonstrated by transformants. Furthermore, the paper made a comparison of crude enzyme catalysis and whole-cell catalysis in an aqueous monophasic system and a water/organic solvent biphasic system. In the water/ n-butyl acetate system, the coexpression system produced 1,398 mM CHBE in the organic phase, which is the highest yield ever reported for CHBE production by NADH-dependent reductases from yeasts. In this case, the molar yield of CHBE was 90.7%, and the total turnover number, defined as moles ( S)-CHBE formed per mole NAD+, was 13,980. [ABSTRACT FROM AUTHOR]
- Subjects :
- BIOSYNTHESIS
CATALYSIS
CARBONYL reductase
GLUCOSE
ENZYMES
ORGANIC solvents
Subjects
Details
- Language :
- English
- ISSN :
- 01757598
- Volume :
- 88
- Issue :
- 6
- Database :
- Complementary Index
- Journal :
- Applied Microbiology & Biotechnology
- Publication Type :
- Academic Journal
- Accession number :
- 55199532
- Full Text :
- https://doi.org/10.1007/s00253-010-2836-4