title: Computational Identification of Amino-Acid Mutations that Further Improve the Activity of a Chalcone-Flavonone Isomerase fromGlycine max.
abstract: Protein design for improving enzymatic activity remains a challenge in biochemistry, especially to identify target amino-acid sites for mutagenesis and to design beneficial mutations for those sites. Here, we employ a computational approach that combines multiple sequence alignment, positive selection detection, and molecular docking to identify and design beneficial amino-acid mutations that further improve the intramolecular-cyclization activity of a chalcone-flavonone isomerase fromGlycine max(GmCHI). By this approach, two GmCHI mutants with higher activities were predicted and verified. The results demonstrate that this approach could determine the beneficial amino-acid mutations for improving the enzymatic activity, and may find more applications in engineering of enzymes.
authors.affiliation: Ministry of Education Key Laboratory for Biodiversity Science and Ecological Engineering, School of Life Sciences, Fudan University Shanghai, China.
authors.forename: Hui
authors.initials: H
authors.lastname: Yuan
authors.affiliation: Ministry of Education Key Laboratory for Biodiversity Science and Ecological Engineering, School of Life Sciences, Fudan University Shanghai, China.
authors.forename: Jiaqi
authors.initials: J
authors.lastname: Wu
authors.affiliation: Department of Biological Sciences, University of North Texas, Denton TX, USA.
authors.forename: Xiaoqiang
authors.initials: X
authors.lastname: Wang
authors.affiliation: Ministry of Education Key Laboratory for Biodiversity Science and Ecological Engineering, School of Life Sciences, Fudan University Shanghai, China.
authors.forename: Jiakuan
authors.initials: J
authors.lastname: Chen
authors.affiliation: Ministry of Education Key Laboratory for Biodiversity Science and Ecological Engineering, School of Life Sciences, Fudan UniversityShanghai, China; Institute of Biodiversity Science and Geobiology, Tibet UniversityLhasa, China.
authors.forename: Yang
authors.initials: Y
authors.lastname: Zhong
authors.affiliation: State Key Laboratory of Genetic Engineering, School of Life Sciences, Fudan University Shanghai, China.
authors.forename: Qiang
authors.initials: Q
authors.lastname: Huang
authors.affiliation: Ministry of Education Key Laboratory for Biodiversity Science and Ecological Engineering, School of Life Sciences, Fudan University Shanghai, China.
authors.forename: Peng
authors.initials: P
authors.lastname: Nan
keywords: chalcone–flavonone isomerase; enzyme engineering; molecular modeling; positive selection; protein design