title: Enzymatic reduction of levulinic acid by engineering the substrate specificity of 3-hydroxybutyrate dehydrogenase.
abstract: Enzymatic reduction of levulinic acid (LA) was performed for the synthesis of 4-hydroxyvaleric acid (4HV)--a monomer of bio-polyester and a precursor of bio-fuels--using 3-hydroxybutyrate dehydrogenase (3HBDH) from Alcaligenes faecalis. Due to the catalytic inactivity of the wild-type enzyme toward LA, engineering of the substrate specificity of the enzyme was performed. A rational design approach with molecular docking simulation was applied, and a double mutant, His144Leu/Trp187Phe, which has catalytic activity (kcat/Km=578.0 min(-1) M(-1)) toward LA was generated. Approximately 57% conversion of LA to 4HV was achieved with this double mutant in 24 h, while no conversion was achieved with the wild-type enzyme.
authors.affiliation: School of Chemical and Biological Engineering, Seoul National University, Seoul 151-742, Republic of Korea.
authors.forename: Young Joo
authors.initials: YJ
authors.lastname: Yeon
authors.forename: Hyung-Yeon
authors.initials: HY
authors.lastname: Park
authors.forename: Young Je
authors.initials: YJ
authors.lastname: Yoo
mesh_terms: D000421:Alcaligenes / Q000201:enzymology*; D006884:Hydroxybutyrate Dehydrogenase / Q000378:metabolism*; D007700:Kinetics; D007982:Levulinic Acids / Q000378:metabolism*; D062105:Molecular Docking Simulation; D050505:Mutant Proteins / Q000378:metabolism; D009154:Mutation / Q000235:genetics; D010084:Oxidation-Reduction; D015202:Protein Engineering / Q000379:methods*; D013379:Substrate Specificity; D014631:Valerates / Q000378:metabolism
chemical_list: D007982:Levulinic Acids; D050505:Mutant Proteins; D014631:Valerates; C469548:4-hydroxyvaleric acid; D006884:Hydroxybutyrate Dehydrogenase; C032246:levulinic acid