title: Enhancement of the catalytic activity of ferulic acid decarboxylase from Enterobacter sp. Px6-4 through random and site-directed mutagenesis.
abstract: The enzyme ferulic acid decarboxylase (FADase) from Enterobacter sp. Px6-4 catalyzes the decarboxylation reaction of lignin monomers and phenolic compounds such as p-coumaric acid, caffeic acid, and ferulic acid into their corresponding 4-vinyl derivatives, that is, 4-vinylphenol, 4-vinylcatechol, and 4-vinylguaiacol, respectively. Among various ferulic acid decarboxylase enzymes, we chose the FADase from Enterobacter sp. Px6-4, whose crystal structure is known, and produced mutants to enhance its catalytic activity by random and site-directed mutagenesis. After three rounds of sequential mutations, FADase(F95L/D112N/V151I) showed approximately 34-fold higher catalytic activity than wild-type for the production of 4-vinylguaiacol from ferulic acid. Docking analyses suggested that the increased activity of FADase(F95L/D112N/V151I) could be due to formation of compact active site compared with that of the wild-type FADase. Considering the amount of phenolic compounds such as lignin monomers in the biomass components, successfully bioengineered FADase(F95L/D112N/V151I) from Enterobacter sp. Px6-4 could provide an ecofriendly biocatalytic tool for producing diverse styrene derivatives from biomass.
authors.affiliation: School of Environmental Science and Engineering, Gwangju Institute of Science and Technology, Gwangju, 500-712, South Korea.
authors.forename: Hyunji
authors.initials: H
authors.lastname: Lee
authors.affiliation: School of Environmental Science and Engineering, Gwangju Institute of Science and Technology, Gwangju, 500-712, South Korea.
authors.forename: Jiyoung
authors.initials: J
authors.lastname: Park
authors.affiliation: School of Environmental Science and Engineering, Gwangju Institute of Science and Technology, Gwangju, 500-712, South Korea.
authors.forename: Chaewon
authors.initials: C
authors.lastname: Jung
authors.affiliation: Max-Planck Institute for Terrestrial Microbiology, Karl-von-Frisch-Street 10, D35043, Marburg, Germany.
authors.forename: Dongfei
authors.initials: D
authors.lastname: Han
authors.affiliation: Radiation Research Division of Industry and Environment, Korea Atomic Energy Research Institute, 1266, Jeongup, 580-185, South Korea.
authors.forename: Jiyoung
authors.initials: J
authors.lastname: Seo
authors.affiliation: Department of Bioscience and Biotechnology, Konkuk University, Seoul, 143-701, South Korea.
authors.forename: Joong-Hoon
authors.initials: JH
authors.lastname: Ahn
authors.affiliation: Department of Bioscience and Biotechnology, Konkuk University, Seoul, 143-701, South Korea.
authors.forename: Youhoon
authors.initials: Y
authors.lastname: Chong
authors.affiliation: School of Environmental Science and Engineering, Gwangju Institute of Science and Technology, Gwangju, 500-712, South Korea. hghur@gist.ac.kr.
authors.forename: Hor-Gil
authors.initials: HG
authors.lastname: Hur
mesh_terms: D055162:Biocatalysis; D001711:Biotransformation; D002262:Carboxy-Lyases / Q000235:genetics* / Q000378:metabolism*; D020134:Catalytic Domain; D003373:Coumaric Acids / Q000378:metabolism; D003653:Decarboxylation; D004754:Enterobacter / Q000201:enzymology* / Q000235:genetics*; D006139:Guaiacol / Q000031:analogs & derivatives / Q000378:metabolism; D062105:Molecular Docking Simulation; D016297:Mutagenesis, Site-Directed; D009154:Mutation; D010636:Phenols / Q000378:metabolism; D011422:Propionates
chemical_list: D003373:Coumaric Acids; D010636:Phenols; D011422:Propionates; D006139:Guaiacol; C014245:4-vinylguaiacol; C004999:ferulic acid; D002262:Carboxy-Lyases; C087194:phenylacrylic acid decarboxylase; C495469:p-coumaric acid; C030626:4-vinylphenol
keywords: 4-Vinylguaiacol; Biotransformation; Directed mutagenesis; Ferulic acid; Ferulic acid decarboxylase