title: Overproduction and substrate specificity of 3-isopropylmalate dehydrogenase from Thiobacillus ferrooxidans.
abstract: We constructed an overexpression system in Escherichia coli of the leuB gene coding for 3-isopropylmalate dehydrogenase in Thiobacillus ferrooxidans. E. coli harboring the plasmid we constructed, pKK leuB1, produced 17-fold the enzyme protein of the expression system previously used for purification. The substrate specificity of the enzyme was analyzed with synthetic (2R, 3S)-3-alkylmalates. The 3-isopropylmalate dehydrogenase of Thiobacillus ferrooxidans had broad specificity toward the alkylmalates.
authors.affiliation: Department of Bioresources Chemistry, Faculty of Agriculture, Okayama University, Japan.
authors.forename: H
authors.initials: H
authors.lastname: Matsunami
authors.forename: H
authors.initials: H
authors.lastname: Kawaguchi
authors.forename: K
authors.initials: K
authors.lastname: Inagaki
authors.forename: T
authors.initials: T
authors.lastname: Eguchi
authors.forename: K
authors.initials: K
authors.lastname: Kakinuma
authors.forename: H
authors.initials: H
authors.lastname: Tanaka
mesh_terms: D050539:3-Isopropylmalate Dehydrogenase; D000429:Alcohol Oxidoreductases / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D015964:Gene Expression Regulation, Bacterial; D007700:Kinetics; D008293:Malates / Q000378:metabolism*; D010957:Plasmids / Q000737:chemistry; D013379:Substrate Specificity; D013499:Surface Properties; D013855:Thiobacillus / Q000201:enzymology* / Q000235:genetics
chemical_list: D008293:Malates; D000429:Alcohol Oxidoreductases; D050539:3-Isopropylmalate Dehydrogenase