title: Characterization of two isozymes of coniferyl alcohol dehydrogenase from Streptomyces sp. NL15-2K.
abstract: We purified two isozymes of coniferyl alcohol dehydrogenase (CADH I and II) to homogeneity from cell-free extracts of Streptomyces sp. NL15-2K. The apparent molecular masses of CADH I and II were determined to be 143 kDa and 151 kDa respectively by gel filtration, whereas their subunit molecular masses were determined to be 35,782.2 Da and 37,597.7 Da respectively by matrix-assisted laser-desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS). Thus, it is probable that both isozymes are tetramers. The optimum pH and temperature for coniferyl alcohol dehydrogenase activity were pH 9.5 and 45 °C for CADH I and pH 8.5 and 40 °C for CADH II. CADH I oxidized various aromatic alcohols and allyl alcohol, and was most efficient on cinnamyl alcohol, whereas CADH II exhibited high substrate specificity for coniferyl alcohol, and showed no activity as to the other alcohols, except for cinnamyl alcohol and 3-(4-hydroxy-3-methoxyphenyl)-1-propanol. In the presence of NADH, CADH I and II reduced cinnamaldehyde and coniferyl aldehyde respectively to the corresponding alcohols.
authors: Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Yasuda Women's University. m-nishi@yasuda-u.ac.jp
authors: Motohiro
authors: M
authors: Nishimura
authors: Kunie
authors: K
authors: Kohno
authors: Yoshio
authors: Y
authors: Nishimura
authors: Masanori
authors: M
authors: Inagaki
authors: Julian
authors: J
authors: Davies
mesh_terms: D000171:Acrolein / Q000031:analogs & derivatives; D000429:Alcohol Oxidoreductases / Q000737:chemistry / Q000378:metabolism*; D000447:Aldehydes / Q000378:metabolism; D001426:Bacterial Proteins / Q000737:chemistry / Q000378:metabolism*; D002457:Cell Extracts / Q000737:chemistry; D002847:Chromatography, Agarose; D006863:Hydrogen-Ion Concentration; D007527:Isoenzymes / Q000737:chemistry / Q000378:metabolism*; D007700:Kinetics; D008970:Molecular Weight; D009243:NAD / Q000378:metabolism; D010636:Phenols / Q000378:metabolism*; D020005:Propanols / Q000378:metabolism*; D021122:Protein Subunits / Q000737:chemistry / Q000378:metabolism*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013302:Streptomyces / Q000737:chemistry / Q000201:enzymology*; D013379:Substrate Specificity; D013696:Temperature
chemical_list: D000447:Aldehydes; D001426:Bacterial Proteins; D002457:Cell Extracts; D007527:Isoenzymes; D010636:Phenols; D020005:Propanols; D021122:Protein Subunits; C075384:coniferaldehyde; D009243:NAD; C006463:allyl alcohol; D000171:Acrolein; C010559:coniferyl alcohol; D000429:Alcohol Oxidoreductases; C019931:coniferyl alcohol dehydrogenase; C020722:cinnamyl alcohol