Genome Mining and Characterization of Diaryl Ketone Reductase from Kluyveromyces polysporus

doi:10.3969/j.issn.1673-1689.2018.03.003

Home > Archive>Volume 37, Issue 3, 2018 >240-249. DOI:10.3969/j.issn.1673-1689.2018.03.003

Genome Mining and Characterization of Diaryl Ketone Reductase from Kluyveromyces polysporus
DOI:
                        10.3969/j.issn.1673-1689.2018.03.003
                    
CSTR:
                        [cstr]
                    
Author:
                        
                        
                    
Affiliation:
Clc Number:Q554；Q781
Fund Project:

Article

Figures

Metrics

Reference

Cited by

Materials

Comments

Abstract:

In this study，an alcohol dehydrogenase KpADH from Kluyveromyces polysporus was identified by genome mining strategy. This newly mined KpADH belongs to extended SDRs subfamily，and catalyze the asymmetric reduction of diaryl ketones into chiral diaryl alcohols. KpADH also displays substrate-coupled NADPH recycling ability by oxidation of cosubstrate isopropanol. The recombinant KpADH was purified by Ni-NTA affinity chromatography，and the enzymatic properties of the purified KpADH were characterized. The optimum pH of oxidizing and reducing activity is 9.5 and 5.5 respectively. The Km and Vmax of KpADH towards CPMK are 0.5 mmol/L and 25 μmol/（min·mg） respectively，while the Km and Vmax of KpADH towards isopropanol are 6.36 mmol/L and 21.37 μmol/（min·mg） respectively. KpADH shows higher catalytic activity on ketone esters and 2，3-butanediol. Within 10 h，100 mmol/L CPMK was asymmetrically reduced into（R）-CPMA by recombinant E. coli BL21/pET28a-kpadh cells，giving >99% conversion，82% ee and 88.7% molar isolation yield. This study provides useful guidance for the application of this newly identified KpADH in the preparation of chiral diaryl alcohols.

Reference

Cited by

Get Citation

TANG Minghui, XU Guochao, NI Ye. Genome Mining and Characterization of Diaryl Ketone Reductase from Kluyveromyces polysporus[J]. Journal of Food Science and Biotechnology,2018,37(3):240-249.

Copy

Article Metrics

Abstract:
PDF:
HTML:
Cited by:

History

Received:
Revised:
Adopted:
Online: June 08,2018
Published:

Article QR Code

Copy Right：Editorial Board of Journal of Food Science and Biotechnology

Address：No. 1800, Lihu Avenue, Wuxi 214122, Jiangsu Province,China PostCode：214122

Phone：0510-85913526 E-mail：xbbjb@jiangnan.edu.cn

Supported by:Beijing E-Tiller Technology Development Co., Ltd.

Get Citation

Share

Article Metrics

History

Article QR Code

WeChat

Mobile website