Highly Active Mutants of Carbonyl Reductase S1 with Inverted Coenzyme Specificity and Production of Optically Active Alcohols

2005 ◽  
Vol 69 (3) ◽  
pp. 544-552 ◽  
Author(s):  
Souichi MORIKAWA ◽  
Takahisa NAKAI ◽  
Yoshihiko YASOHARA ◽  
Hirokazu NANBA ◽  
Noriyuki KIZAKI ◽  
...  
Keyword(s):  
Carbonyl Reductase ◽  
Optically Active ◽  
Coenzyme Specificity ◽  
Highly Active

Access to Optically Active Aryl Halohydrins Using a Substrate-Tolerant Carbonyl Reductase Discovered from Kluyveromyces thermotolerans

ACS Catalysis ◽  
2012 ◽  
Vol 2 (12) ◽  
pp. 2566-2571 ◽  
Author(s):  
Guo-Chao Xu ◽  
Hui-Lei Yu ◽  
Xiao-Yan Zhang ◽  
Jian-He Xu
Keyword(s):  
Carbonyl Reductase ◽  
Optically Active

ChemInform Abstract: Synthesis of Optically Active α-Bromohydrins via Reduction of α-Bromoacetophenone Analogues Catalyzed by an Isolated Carbonyl Reductase.

ChemInform ◽  
2012 ◽  
Vol 43 (41) ◽  
pp. no-no
Author(s):  
Jie Ren ◽  
Wenyue Dong ◽  
Benqing Yu ◽  
Qiaqing Wu ◽  
Dunming Zhu
Keyword(s):  
Carbonyl Reductase ◽  
Optically Active

Probing Structure-Property Relationships in a Family of Aryloxyphenoxy Propionate Herbicides Using Single Crystal X-Ray Diffraction Techniques

1991 ◽  
Vol 35 (A) ◽  
pp. 641-643
Author(s):  
Philip R. Rudolf ◽  
Larry Kershner ◽  
Jim Tai
Keyword(s):  
General Structure ◽  
Optically Active ◽  
Herbicidal Activity ◽  
Structure Property ◽  
X Ray Diffraction ◽  
Methyl Lactate ◽  
X Ray ◽  
Highly Active ◽  
Structure Property Relationships ◽  
Sulfonate Ester

Aryloxyphenoxy propionates are a widely manufactured class of highly active grass-selective herbicides for use in various crops. Examples include VERDICT (1a), FUSILADE (1b), WHIP (1c), and ASSURE (1d). Figure 1 shows the general structure of aryloxyphenoxy propionates. All of these compounds contain a chiral center and each enantiomer of the optical pair exhibits significantly different herbicidal activity. The optically active isomers can be prepared by reacting the corresponding substituted phenoxyphenol with the S-(-)methyl lactate sulfonate ester (2). However, the optical yield from this type of reaction is usually less than 85%. Optical purification can be achieved by control of the crystallization.

scholarly journals Ser67Asp and His68Asp Substitutions in Candida parapsilosis Carbonyl Reductase Alter the Coenzyme Specificity and Enantioselectivity of Ketone Reduction

2009 ◽  
Vol 75 (7) ◽  
pp. 2176-2183 ◽  
Author(s):  
Rongzhen Zhang ◽  
Yan Xu ◽  
Ying Sun ◽  
Wenchi Zhang ◽  
Rong Xiao
Keyword(s):  
Candida Parapsilosis ◽  
Enzyme Stability ◽  
Optical Purity ◽  
Carbonyl Reductase ◽  
Site Directed Mutagenesis ◽  
Short Chain ◽  
Significant Shift ◽  
Wild Type ◽  
Ketone Reduction ◽  
Coenzyme Specificity

ABSTRACT A short-chain carbonyl reductase (SCR) from Candida parapsilosis catalyzes an anti-Prelog reduction of 2-hydroxyacetophenone to (S)-1-phenyl-1,2-ethanediol (PED) and exhibits coenzyme specificity for NADPH over NADH. By using site-directed mutagenesis, the mutants were designed with different combinations of Ser67Asp, His68Asp, and Pro69Asp substitutions inside or adjacent to the coenzyme binding pocket. All mutations caused a significant shift of enantioselectivity toward the (R)-configuration during 2-hydroxyacetophenone reduction. The S67D/H68D mutant produced (R)-PED with high optical purity and yield in the NADH-linked reaction. By kinetic analysis, the S67D/H68D mutant resulted in a nearly 10-fold increase and a 20-fold decrease in the k cat/Km value when NADH and NADPH were used as the cofactors, respectively, but maintaining a k cat value essentially the same with respect to wild-type SCR. The ratio of Kd (dissociation constant) values between NADH and NADPH for the S67D/H68D mutant and SCR were 0.28 and 1.9 respectively, which indicates that the S67D/H68D mutant has a stronger preference for NADH and weaker binding for NADPH. Moreover, the S67D/H68D enzyme exhibited a secondary structure and melting temperature similar to the wild-type form. It was also found that NADH provided maximal protection against thermal and urea denaturation for S67D/H68D, in contrast to the effective protection by NADP(H) for the wild-type enzyme. Thus, the double point mutation S67D/H68D successfully converted the coenzyme specificity of SCR from NADP(H) to NAD(H) as well as the product enantioselectivity without disturbing enzyme stability. This work provides a protein engineering approach to modify the coenzyme specificity and enantioselectivity of ketone reduction for short-chain reductases.

scholarly journals Switch of Coenzyme Specificity of Mouse Lung Carbonyl Reductase by Substitution of Threonine 38 with Aspartic Acid

1997 ◽  
Vol 272 (4) ◽  
pp. 2218-2222 ◽  
Author(s):  
Masayuki Nakanishi ◽  
Kazuya Matsuura ◽  
Hiroyuki Kaibe ◽  
Nobutada Tanaka ◽  
Takamasa Nonaka ◽  
...  
Keyword(s):  
Aspartic Acid ◽  
Carbonyl Reductase ◽  
Mouse Lung ◽  
Coenzyme Specificity

Synthesis of optically active α-bromohydrins via reduction of α-bromoacetophenone analogues catalyzed by an isolated carbonyl reductase

2012 ◽  
Vol 23 (6-7) ◽  
pp. 497-500 ◽  
Author(s):  
Jie Ren ◽  
Wenyue Dong ◽  
Benqing Yu ◽  
Qiaqing Wu ◽  
Dunming Zhu
Keyword(s):  
Carbonyl Reductase ◽  
Optically Active
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