title: Probing role of key residues in the divergent evolution of Yarrowia lipolytica lipase 2 and Aspergillus niger eruloyl esterase A.
abstract: Yarrowia lipolytica lipase 2 (YLLip2) and Aspergillus niger feruloyl esterase A (AnFaeA) are enzymes of similar structures but with different functions. They are both classified into the same homologous family in Lipase Engineering Database (LED). The major difference between the two enzymes is that YLLip2 exhibits interfacial activity while AnFaeA does not. In order to better understand the interfacial activation mechanisms of YLLip2, structure guided site-directed mutagenesis were performed, mutants were constructed, kinetics parameters and lipase properties were detected. Mutant enzymes showed enhanced catalytic efficiency towards p-nitrophenyl butyrin (pNPB) but their catalytic efficiency decreased towards p-nitrophenyl palmitate (pNPP), their catalysis behavior was more close to feruloyl esterase. Moreover, the mutant enzymes exhibited enhanced thermostability compared with their wild type. These results indicate that I100 and F129 are probably cut-off point of divergent functions between the two enzymes during evolution.
authors: Key Laboratory of Molecular Biophysics of the Ministry of Education, College of Life Science and Technology, Huazhong University of Science and Technology, Wuhan, PR China.
authors: Guilong
authors: G
authors: Wang
authors: Key Laboratory of Molecular Biophysics of the Ministry of Education, College of Life Science and Technology, Huazhong University of Science and Technology, Wuhan, PR China.
authors: Zimin
authors: Z
authors: Liu
authors: Key Laboratory of Molecular Biophysics of the Ministry of Education, College of Life Science and Technology, Huazhong University of Science and Technology, Wuhan, PR China.
authors: Li
authors: L
authors: Xu
authors: Key Laboratory of Molecular Biophysics of the Ministry of Education, College of Life Science and Technology, Huazhong University of Science and Technology, Wuhan, PR China.
authors: Houjin
authors: H
authors: Zhang
authors: Key Laboratory of Molecular Biophysics of the Ministry of Education, College of Life Science and Technology, Huazhong University of Science and Technology, Wuhan, PR China; Shenzhen R & D Center of Huazhong University of Science and Technology, Shenzhen, PR China. Electronic address: yanyunjun@hust.edu.cn.
authors: Yunjun
authors: Y
authors: Yan
mesh_terms: D000595:Amino Acid Sequence; D001234:Aspergillus niger / Q000201:enzymology*; D002087:Butyrates / Q000378:metabolism; D002265:Carboxylic Ester Hydrolases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D004252:DNA Mutational Analysis; D004795:Enzyme Stability; D007700:Kinetics; D008049:Lipase / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D050505:Mutant Proteins / Q000235:genetics / Q000378:metabolism; D010168:Palmitates / Q000378:metabolism; D011487:Protein Conformation; D016415:Sequence Alignment; D013379:Substrate Specificity; D013696:Temperature; D025062:Yarrowia / Q000201:enzymology*
chemical_list: D002087:Butyrates; D050505:Mutant Proteins; D010168:Palmitates; C050015:4-nitrophenyl palmitate; C033592:4-nitrophenyl butyrate; D002265:Carboxylic Ester Hydrolases; D008049:Lipase; C070196:feruloyl esterase
keywords: Aspergillus niger feruloyl esterase A; Divergent evolution; Interfacial activation; Site-directed mutagenesis; Yarrowia lipolytica lipase 2