Coupled (R)-carbonyl reductase and glucose dehydrogenase catalyzes (R)-1-phenyl-1, 2-ethanediol biosynthesis with excellent stereochemical selectivity. Issue 11 (November 2015)

• Record Type: Journal Article
• Authors: Zhou, Xiaotian; Zhang, Rongzhen; Xu, Yan; Liang, Hongbo; Jiang, Jiawei; Xiao, Rong
• Is Part Of: Process biochemistry. Volume 50:Issue 11(2015:Nov.)
• Page Start: 1807
• Page End: 1813
• Publication Date: 2015-11
• Subjects: GDH glucose dehydrogenase -- 2-HAP 2-hydroxyacetophenone -- RCR (R)-carbonyl reductase -- (R)-PED (R)-1-phenyl-1, 2-ethanediol -- (S)-PED (S)-1-phenyl-1, 2-ethanediol; (R)-1-Phenyl-1, 2-ethanediol -- (R)-Carbonyl reductase -- NADH regeneration -- Glucose dehydrogenase -- Candida parapsilosis; Biochemical engineering -- Periodicals; Biotechnology -- Periodicals; Biochemistry -- periodicals; Biotechnology -- periodicals; Chemical Engineering -- periodicals; Génie biochimique -- Périodiques; Biotechnologie -- Périodiques; Biochemical engineering; Biotechnology; Periodicals; 660.63
• DOI: 10.1016/j.procbio.2015.08.002
• Languages: English
• ISSNs: 1359-5113

Abstract

Graphical abstract: The coupled system based on RCR and GDH was rebalanced through protein-expression optimization. It gave excellent performance on ( R )-PED biotransformation without exogenous cofactors. Highlights: The construction of coupling system contain RCR and GDH. The introduction of GDH has little effects on cell-growth. The functions of RCR and GDH were rebalanced through protein-expression optimization. The coupling system produced ( R )-PED with excellent stereochemical selectivity. Abstract: The biotransformation of 2-hydroxyacetophenone to ( R )-1-phenyl-1, 2-ethanediol (PED) by NADH-dependent ( R )-carbonyl reductase (RCR) from Candida parapsilosis is slow and gives low yields, probably as a result of insufficient cofactors. To improve the biotransformation efficiency of ( R )-PED from 2-hydroxyacetophenon, an enzyme-coupling system containing RCR and glucose dehydrogenase (GDH) was constructed to strengthen NADH-recycling pathway in Escherichia coli, in which the Shine-Dalgarno sequence and the aligned spacing sequence were used as linkers between them. The introduction of glucose dehydrogenase had little affects on the cell-growth. The co-expression conditions of RCR and glucose dehydrogenase was optimized to rebalance their catalytic functions. The ratio of k cat / K M for enzyme-coupling system catalyzing 2-HAP and glucose was about 1.0, suggesting the good balance between the functions of RCR and GDH. The rebalanced system gave excellent performance in ( R )-PED biotransformation: an optical purity of 99.9% and a yield of 99.9% at optimal conditions: 35 °C and pH 7.0. The introduction of glucose dehydrogenase stimulated increases of 23.8% and 63.8%, in optical purity and yield of ( R )-PED, and simultaneously reduced the reaction time two-fold. This work provided a valuable method for efficient chiral alcohol production through protein-expression and biotransformation optimization to rebalance cofactor pathways.