Semi-Rational Engineering a Carbonyl Reductase for the Enantioselective Reduction of beta-Amino Ketones

CORC > 天津工业生物技术研究所 > 生物催化与绿色化工朱敦明 > 期刊论文

	Semi-Rational Engineering a Carbonyl Reductase for the Enantioselective Reduction of beta-Amino Ketones
	Zhang, Dalong 1,2; Chen, Xi 1,2; Chi, Jing 1,2,3; Feng, Jinhui 1,2; Wu, Qiaqing 1,2; Zhu, Dunming 1,2
刊名	ACS CATALYSIS
	2015-04-01
卷号	5 期号:4 页码:2452-2457
关键词	chiral gamma-amino alcohols beta-amino ketones enzyme engineering carbonyl reductase enantioselective reduction
英文摘要	Chiral 3-(dimethylamino)-1-phenylpropan-1-ol (2a) and 3-(dimethylamino)-1-(2-thienyl)-1-ol (2b) are very important intermediates for the synthesis of antidepressants. The mutant M242F/Q245T of a carbonyl reductase (SSCR) from Sporobolomyces salmonicolor AKU4429 catalyzed the reduction of 3-(dimethylamino)-1-phenylpropan-1-one (la) to the (S)-2a (28% ee). The combinatorial active-site saturation of this enzyme resulted in two mutants P170R/L174Y and P170H/L174Y, which catalyzed the reduction of la and 3-(dimethylamino)-1-(2-thienyl)-propan-1-one (1b) to give the (R)-gamma-amino alcohols with up to 95% ee, respectively. The individual site saturation mutagenesis of Pro170 and Leu174 revealed that Pro170 did not significantly affect the enzyme enantioselectivity toward la and 1b, whereas residue Leu174 played a critical role in determining the enantioselectivity. Mutant L174W catalyzed the reduction of la to the (S)-gamma-amino alcohol with increased enantioselectivity from 28% ee to 96% ee, although mutant L174Y exhibited (R)-preference in 88% ee. For 1b, the (R)-alcohol was obtained with 95% ee by using variant L174Y as the catalyst, whereas L174W exhibited (S)-preference in 65% ee. The kinetic studies indicated that catalytic efficiencies (k(cat)/K-m) of these mutants were also improved. The enzyme substrate docking provided some insights into the structural basis for the reversal of enantioselectivity by the substitution of Leu174.
WOS标题词	Science & Technology ; Physical Sciences
类目[WOS]	Chemistry, Physical
研究领域[WOS]	Chemistry
关键词[WOS]	SINGLE-POINT MUTATION ; ASYMMETRIC HYDROGENATION ; SPOROBOLOMYCES-SALMONICOLOR ; DIRECTED EVOLUTION ; SATURATION MUTAGENESIS ; SELECTIVE INHIBITOR ; EPOXIDE HYDROLASE ; SEROTONIN-UPTAKE ; OPTICAL ISOMERS ; SUBSTRATE SCOPE
收录类别	SCI
语种	英语
WOS记录号	WOS:000352464400050
内容类型	期刊论文
源URL	[http://124.16.173.210/handle/834782/1454]
专题	天津工业生物技术研究所_生物催化与绿色化工朱敦明_期刊论文
作者单位	1.Chinese Acad Sci, Tianjin Inst Ind Biotechnol, Natl Engn Lab Ind Enzymes, Tianjin 300308, Peoples R China 2.Chinese Acad Sci, Tianjin Inst Ind Biotechnol, Tianjin Engn Ctr Biocatalyt Technol, Tianjin 300308, Peoples R China 3.Univ Chinese Acad Sci, Beijing 100049, Peoples R China
推荐引用方式 GB/T 7714	Zhang, Dalong,Chen, Xi,Chi, Jing,et al. Semi-Rational Engineering a Carbonyl Reductase for the Enantioselective Reduction of beta-Amino Ketones[J]. ACS CATALYSIS,2015,5(4):2452-2457.
APA	Zhang, Dalong,Chen, Xi,Chi, Jing,Feng, Jinhui,Wu, Qiaqing,&Zhu, Dunming.(2015).Semi-Rational Engineering a Carbonyl Reductase for the Enantioselective Reduction of beta-Amino Ketones.ACS CATALYSIS,5(4),2452-2457.
MLA	Zhang, Dalong,et al."Semi-Rational Engineering a Carbonyl Reductase for the Enantioselective Reduction of beta-Amino Ketones".ACS CATALYSIS 5.4(2015):2452-2457.