The catalytic property of 3-hydroxyisobutyrate dehydrogenase from Bacillus cereus on 3-hydroxypropionate

Appl Biochem Biotechnol. 2010 Mar;160(3):694-703. doi: 10.1007/s12010-009-8685-x. Epub 2009 Jun 11.

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.

MeSH terms

  • Alcohol Oxidoreductases / biosynthesis
  • Alcohol Oxidoreductases / genetics
  • Alcohol Oxidoreductases / isolation & purification
  • Alcohol Oxidoreductases / metabolism*
  • Aldehydes / metabolism
  • Amino Acid Sequence
  • Bacillus cereus / enzymology*
  • Bacillus cereus / genetics
  • Biocatalysis*
  • Cloning, Molecular
  • Enzyme Stability
  • Hydrogen-Ion Concentration
  • Kinetics
  • Lactic Acid / analogs & derivatives*
  • Lactic Acid / metabolism
  • Metals / pharmacology
  • Molecular Sequence Data
  • Spectrophotometry
  • Substrate Specificity
  • Temperature
  • Thiobarbiturates / metabolism

Substances

  • Aldehydes
  • Metals
  • Thiobarbiturates
  • Lactic Acid
  • hydracrylic acid
  • Alcohol Oxidoreductases
  • 3-hydroxyisobutyrate dehydrogenase
  • thiobarbituric acid