title: Purification and characterization of grass carp mitochondrial aldehyde dehydrogenase.
abstract: The molecular biology and enzymology of aldehyde dehydrogenase (ALDH) have been extensively investigated. However, most of the studies have been confined to the mammalian forms, while the sub-mammalian vertebrate ALDHs are relatively unexplored. In the present investigation, an ALDH was purified from the hepatopancreas of grass carp (Ctenopharygodon idellus) by affinity chromatographies on alpha-cyanocinnamate-Sepharose and Affi-gel Blue agarose. The 800-fold purified enzyme had a specific activity of 4.46 U/mg toward the oxidation of acetaldehyde at pH 9.5. It had a subunit molecular weight of 55000. Isoelectric focusing showed a single band with a pI of 5.3. N-terminal amino acid sequencing of 30 residues revealed a positional identity of approximately 70% with mammalian mitochondrial ALDH2. The kinetic properties of grass carp ALDH resembled those of mammalian ALDH2. The optimal pH for the oxidation of acetaldehyde was 9.5. The K(m) values for acetaldehyde were 0.36 and 0.31 microM at pH 7.5 and 9.5, respectively. Grass carp ALDH also possessed esterase activity which could be activated in the presence of NAD(+).
authors: Department of Biochemistry, The Chinese University of Hong Kong, Shatin, N.T., Hong Kong. wpfong@cuhk.edu.hk
authors: W
authors: W
authors: Fong
authors: K
authors: K
authors: Choy
mesh_terms: D000444:Aldehyde Dehydrogenase / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D002347:Carps / Q000235:genetics / Q000378:metabolism*; D002846:Chromatography, Affinity; D004064:Digestive System / Q000201:enzymology; D006801:Humans; D006863:Hydrogen-Ion Concentration; D066298:In Vitro Techniques; D007700:Kinetics; D008928:Mitochondria / Q000201:enzymology; D008969:Molecular Sequence Data; D008970:Molecular Weight; D021122:Protein Subunits; D017386:Sequence Homology, Amino Acid; D013045:Species Specificity
chemical_list: D021122:Protein Subunits; D000444:Aldehyde Dehydrogenase