title: Conversion of a Rhizopus chinensis lipase into an esterase by lid swapping.
abstract: In an effort to explore the feasibility of converting a lipase into an esterase by modifying the lid region, we designed and characterized two novel Rhizopus chinensis lipase variants by lid swapping. The substrate specificity of an R. chinensis lipase was successfully modified toward water-soluble substrates, that is, turned into an esterase, by replacing the hydrophobic lid with a hydrophilic lid from ferulic acid esterase from Aspergillus niger Meanwhile, as a comparison, the lid of R. chinensis lipase was replaced by a hydrophobic lid from Rhizomucor miehei lipase, which did not alter its substrate specificity but led to a 5.4-fold higher catalytic efficiency (k*cat/K*m) toward p-nitrophenyl laurate. Based on the analysis of structure-function relationships, it suggests that the amphipathic nature of the lid is very important for the substrate specificity. This study provides new insight into the structural basis of lipase specificities and a way to tune the substrate preference of lipases.
authors.affiliation: The Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi 214122, Jiangsu, China.
authors.forename: Xiao-Wei
authors.initials: XW
authors.lastname: Yu
authors.affiliation: The Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology.
authors.forename: Shan-Shan
authors.initials: SS
authors.lastname: Zhu
authors.affiliation: Center for Advanced Biotechnology and Medicine, Department of Molecular Biology and Biochemistry, Rutgers University, Piscataway, NJ 08854.
authors.forename: Rong
authors.initials: R
authors.lastname: Xiao
authors.affiliation: The Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi 214122, Jiangsu, China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Xu
mesh_terms: D001234:Aspergillus niger / Q000201:enzymology / Q000235:genetics; D002265:Carboxylic Ester Hydrolases / Q000737:chemistry / Q000235:genetics; D005656:Fungal Proteins / Q000737:chemistry / Q000235:genetics; D008049:Lipase / Q000737:chemistry / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics; D012233:Rhizopus / Q000201:enzymology / Q000235:genetics
chemical_list: D005656:Fungal Proteins; D011993:Recombinant Fusion Proteins; D002265:Carboxylic Ester Hydrolases; D008049:Lipase; C070196:feruloyl esterase
keywords: chain-length specificity • chimera • domain exchange