Development of a whole-cell biocatalyst with NADPH regeneration system for biosulfoxidation

• Published in: Journal of industrial microbiology & biotechnology 2013-08, Vol.40 (8), p.797-803
• Main Authors: Zhai, Xiao-Hong, Ma, Yuan-Hui, Lai, Dun-Yue, Zhou, Shuo, Chen, Zhen-Ming
• Format: Article
• Language: English
• Subjects: Acinetobacter calcoaceticus; Acinetobacter calcoaceticus - enzymology; Analysis; Antibiotics; Bacteria; Biocatalysis; Biocatalysts; Biochemistry; Bioinformatics; Biological and medical sciences; Biomedical and Life Sciences; Biotechnology; Candida - enzymology; Candida boidinii; Dehydrogenases; E coli; enantiomers; Enzymes; Escherichia coli; Escherichia coli - genetics; Escherichia coli - metabolism; Formate Dehydrogenases - genetics; Formate Dehydrogenases - metabolism; Fundamental and applied biological sciences. Psychology; Gene expression; genes; Genetic Engineering; Inorganic Chemistry; Life Sciences; Microbiology; NADP (coenzyme); NADP - metabolism; Optimization; Oxidation; Oxidation-Reduction; Oxygenases - genetics; Oxygenases - metabolism; Plasmids; Proteins; Studies; Sulfoxides - metabolism; Yeast
• ISSN: 1367-5435; 1476-5535
• DOI: 10.1007/s10295-013-1288-0

A formate dehydrogenase gene (fdh) originated from Candida boidinii was co-expressed in E. coli BL21 (DE3) with the cyclohexanone monooxygenase gene (chmo) cloned from Acinetobacter calcoaceticus NCIMB 9871. The co-expression system was then used as a whole-cell biocatalyst to synthesize chiral phenyl methyl sulfoxide (PMSO) from thioanisole (PMS) and the reaction conditions were investigated. When the initial concentration of PMS was 20 mM, the specific productivity of PMSO in this system was 2.07 μmol g⁻¹ cw min⁻¹ (cw: wet cell weight) and the ee value for the R-sulfoxide was 99 %. In contrast, when chmo was the only gene expressed in E. coli, the specific productivity of PMSO was 0.053 μmol g⁻¹ cw min⁻¹ with no exact enantioselectivity. Further determination of NADPH concentration in the whole-cell catalysts suggested that co-expression of fdh with chmo significantly improved NADPH supply. Thus, this whole-cell biocatalyst system is highly advantageous for the synthesis of optically pure R-sulfoxide.