title: The catalytic property of 3-hydroxyisobutyrate dehydrogenase from Bacillus cereus on 3-hydroxypropionate.
abstract: The MmsB gene product from Bacillus cereus ATCC14579 exhibits 3-hydroxypropionate dehydrogenase activity. It encodes the 32-kDa enzyme protein composed of 292 amino acids. Recombinant 3-hydroxyisobutyrate dehydrogenase (3-HIBADH) was purified 100-fold from cell extract by ammonium sulfate fractionation and column chromatography. The enzyme catalyzed oxidation of 3-hydroxypropionate (3-HP) between pH 7.0 and 10.0 with optimal activity between 8.8 and 9.0. A Km of 16.8 mM for 3-HP was calculated from a Lineweaver-Burk plot. The semialdehyde as products has been proven by spectrophotometric determination. The dehydrogenase apparently has no metal ion requirement. Kinetic determinations established that 3-HIBADH was more active with NADP(+) than NAD(+), which did not show similarity with previously reported 3-HIBADH except that from Thermus thermophilus.
authors.affiliation: Nanjing University of Technology, Nanjing, China.
authors.forename: Tianran
authors.initials: T
authors.lastname: Yao
authors.forename: Lin
authors.initials: L
authors.lastname: Xu
authors.forename: Hanjie
authors.initials: H
authors.lastname: Ying
authors.forename: He
authors.initials: H
authors.lastname: Huang
authors.forename: Ming
authors.initials: M
authors.lastname: Yan
mesh_terms: D000429:Alcohol Oxidoreductases / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D000447:Aldehydes / Q000378:metabolism; D000595:Amino Acid Sequence; D001409:Bacillus cereus / Q000201:enzymology* / Q000235:genetics; D055162:Biocatalysis; D003001:Cloning, Molecular; D004795:Enzyme Stability; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D019344:Lactic Acid / Q000031:analogs & derivatives* / Q000378:metabolism; D008670:Metals / Q000494:pharmacology; D008969:Molecular Sequence Data; D013053:Spectrophotometry; D013379:Substrate Specificity; D013696:Temperature; D013858:Thiobarbiturates / Q000378:metabolism
chemical_list: D000447:Aldehydes; D008670:Metals; D013858:Thiobarbiturates; D019344:Lactic Acid; C031601:hydracrylic acid; D000429:Alcohol Oxidoreductases; C021398:3-hydroxyisobutyrate dehydrogenase; C029684:thiobarbituric acid