Abstract
A novel aldo–keto reductase (LEK) from Lodderomyces elongisporus NRRL YB-4239 (ATCC 11503) was discovered by genome database mining for carbonyl reduction. LEK was overexpressed in Escherichia coli BL21 (DE3), purified to homogeneity and the catalytic properties were studied. Among the substrates, ethyl 4-chloro-3-oxobutanoate was converted to ethyl (R)-4-chloro-3- hydroxybutanoate ((R)-CHBE), an important pharmaceutical intermediate, with an excellent enantiomeric excess (e.e.) (>99 %). The mutants W28A and S209G obtained by site-directed mutation were identified with much higher molar conversion yields and lower Km values. Further, the constructed coenzyme regeneration system with glucose as co-substrate resulted in a yield of 100 %, an enantioselectivity of >99 %, and the calculated production rate of 56.51 mmol/L/H. These results indicated the potential of LEK for the industrial production of (R)-CHBE and other valuable chiral alcohols.
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Acknowledgments
The work was supported by the Technology Research and Development Program of Hangzhou (20130432B05, 20120232B13, and 20130533B17), Zhejiang Provincial Natural Science Foundation of China (LY13C050004, LQ12B06007, and LY13B060008), National Natural Science Foundation of China (21006018, 30900253, and 21206024).
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The experiments comply with the current laws of China in which they were performed.
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Wang, Q., Ye, T., Ma, Z. et al. Characterization and site-directed mutation of a novel aldo–keto reductase from Lodderomyces elongisporus NRRL YB-4239 with high production rate of ethyl (R)-4-chloro-3-hydroxybutanoate. J Ind Microbiol Biotechnol 41, 1609–1616 (2014). https://doi.org/10.1007/s10295-014-1502-8
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DOI: https://doi.org/10.1007/s10295-014-1502-8