Rational design of Kluyveromyces marxianus ZJB14056 aldo-keto reductase KmAKR to enhance diastereoselectivity and activity. 2017

Ya-Jun Wang, and Bin-Bin Ying, and Wei Shen, and Ren-Chao Zheng, and Yu-Guo Zheng
Key Laboratory of Bioorganic Synthesis of Zhejiang Province, College of Biotechnology and Bioengineering, Zhejiang University of Technology, Hangzhou, Zhejiang, 310014, PR China; Engineering Research Center of Bioconversion and Biopurification of the Ministry of Education, Zhejiang University of Technology, Hangzhou, Zhejiang, 310014, PR China.

t-Butyl 6-cyano-(3R,5R)-dihydroxyhexanoate ((3R,5R)-1b) is a valuable chiral synthon of atorvastatin calcium. A novel NADPH-specific aldo-keto reductase (AKR) was identified from a thermotolerant yeast Kluyveromyces marxianus ZJB14056 by genome database mining, displaying t-butyl 6-cyano-(5R)-hydroxy-3-oxohexanoate ((5R)-1a) reducing activity and moderate diastereoselectivity (dep∼80.5%). Molecular homology modeling and docking studies demonstrated that the side chain of Trp297 blocks binding of (5R)-1a to KmAKR. The mutation of Trp297 to His led to dramatic conformational changes and significant improvement in both diastereoselectivity and activity. In comparison with KmAKR, KmAKR-W297H displayed strict diastereoselectivity, and 2.8-fold, 3.9-fold improvement in kcat and kcat/Km toward (5R)-1a, which were 10.36s-1 and 6.56s-1·mM-1 respectively. Coupling KmAKR-W297H with Exiguobacterium sibiricum glucose dehydrogenase (EsGDH) for coenzyme regeneration, 100mM (5R)-1a was completely reduced to (3R,5R)-1b within 12h, in a dep >99.5%.

UI MeSH Term Description Entries
D007700 Kinetics The rate dynamics in chemical or physical systems.
D007716 Kluyveromyces An ascomycetous yeast of the fungal family Saccharomycetaceae, order SACCHAROMYCETALES. Kluyveromyce
D004795 Enzyme Stability The extent to which an enzyme retains its structural conformation or its activity when subjected to storage, isolation, and purification or various other physical or chemical manipulations, including proteolytic enzymes and heat. Enzyme Stabilities,Stabilities, Enzyme,Stability, Enzyme
D005656 Fungal Proteins Proteins found in any species of fungus. Fungal Gene Products,Fungal Gene Proteins,Fungal Peptides,Gene Products, Fungal,Yeast Proteins,Gene Proteins, Fungal,Peptides, Fungal,Proteins, Fungal
D005800 Genes, Fungal The functional hereditary units of FUNGI. Fungal Genes,Fungal Gene,Gene, Fungal
D006863 Hydrogen-Ion Concentration The normality of a solution with respect to HYDROGEN ions; H+. It is related to acidity measurements in most cases by pH pH,Concentration, Hydrogen-Ion,Concentrations, Hydrogen-Ion,Hydrogen Ion Concentration,Hydrogen-Ion Concentrations
D000069059 Atorvastatin A pyrrole and heptanoic acid derivative, HYDROXYMETHYLGLUTARYL-COA REDUCTASE INHIBITOR (statin), and ANTICHOLESTEREMIC AGENT that is used to reduce serum levels of LDL-CHOLESTEROL; APOLIPOPROTEIN B; and TRIGLYCERIDES. It is used to increase serum levels of HDL-CHOLESTEROL in the treatment of HYPERLIPIDEMIAS, and for the prevention of CARDIOVASCULAR DISEASES in patients with multiple risk factors. (3R,5R)-7-(2-(4-Fluorophenyl)-5-isopropyl-3-phenyl-4-(phenylcarbamoyl)-1H-pyrrol-1-yl)-3,5-dihydroxyheptanoic acid,Atorvastatin Calcium,Atorvastatin Calcium Anhydrous,Atorvastatin Calcium Hydrate,Atorvastatin Calcium Trihydrate,Atorvastatin, Calcium Salt,CI 981,CI-981,Lipitor,Liptonorm,CI981
D000074408 Aldo-Keto Reductases A family of NADPH-dependent oxidoreductases that reduce carbonyl substrates including sugar-aldehydes, KETOSTEROIDS; keto-prostaglandins, and QUINONES. They are monomers of approximately 37 KDa and are characterized by a parallel beta-8 (BETA SHEET)-alpha 8 (ALPHA HELICES)-barrel structure that contains the NADP binding site. This conformation favors aromatic and apolar substrates. Aldo-Keto Reductase,Aldo-Keto Reductase Family,Aldo-Keto Reductase Superfamily,Aldo Keto Reductase,Aldo Keto Reductase Family,Aldo Keto Reductase Superfamily,Aldo Keto Reductases,Reductase, Aldo-Keto,Reductases, Aldo-Keto
D000595 Amino Acid Sequence The order of amino acids as they occur in a polypeptide chain. This is referred to as the primary structure of proteins. It is of fundamental importance in determining PROTEIN CONFORMATION. Protein Structure, Primary,Amino Acid Sequences,Sequence, Amino Acid,Sequences, Amino Acid,Primary Protein Structure,Primary Protein Structures,Protein Structures, Primary,Structure, Primary Protein,Structures, Primary Protein
D013237 Stereoisomerism The phenomenon whereby compounds whose molecules have the same number and kind of atoms and the same atomic arrangement, but differ in their spatial relationships. (From McGraw-Hill Dictionary of Scientific and Technical Terms, 5th ed) Molecular Stereochemistry,Stereoisomers,Stereochemistry, Molecular,Stereoisomer

Related Publications

Ya-Jun Wang, and Bin-Bin Ying, and Wei Shen, and Ren-Chao Zheng, and Yu-Guo Zheng
April 2016, Journal of biotechnology,
Ya-Jun Wang, and Bin-Bin Ying, and Wei Shen, and Ren-Chao Zheng, and Yu-Guo Zheng
September 2019, Bioorganic chemistry,
Ya-Jun Wang, and Bin-Bin Ying, and Wei Shen, and Ren-Chao Zheng, and Yu-Guo Zheng
March 2024, Biotechnology journal,
Ya-Jun Wang, and Bin-Bin Ying, and Wei Shen, and Ren-Chao Zheng, and Yu-Guo Zheng
November 2021, Biotechnology and bioengineering,
Ya-Jun Wang, and Bin-Bin Ying, and Wei Shen, and Ren-Chao Zheng, and Yu-Guo Zheng
October 2020, Bioorganic chemistry,
Ya-Jun Wang, and Bin-Bin Ying, and Wei Shen, and Ren-Chao Zheng, and Yu-Guo Zheng
September 2021, Biotechnology journal,
Ya-Jun Wang, and Bin-Bin Ying, and Wei Shen, and Ren-Chao Zheng, and Yu-Guo Zheng
June 2012, Nihon yakurigaku zasshi. Folia pharmacologica Japonica,
Ya-Jun Wang, and Bin-Bin Ying, and Wei Shen, and Ren-Chao Zheng, and Yu-Guo Zheng
March 2015, The Journal of biological chemistry,
Ya-Jun Wang, and Bin-Bin Ying, and Wei Shen, and Ren-Chao Zheng, and Yu-Guo Zheng
October 2023, Microbial cell factories,
Ya-Jun Wang, and Bin-Bin Ying, and Wei Shen, and Ren-Chao Zheng, and Yu-Guo Zheng
April 2021, Bioorganic chemistry,
Copied contents to your clipboard!