Purification and characterization of vanillin dehydrogenases from alkaliphile Micrococcus sp. TA1 and neutrophile Burkholderia cepacia TM1.
Vanillin dehydrogenases (VDHs) were purified and characterized from two bacterial strains that have different pH dependencies for growth. The alkaliphile Micrococcus sp. TA1, isolated from an alkaline spa, can grow on several aromatic compounds such as ferulic acid, vanillin, vanillic acid, and protocatechuic acid under alkaline conditions. The neutrophile Burkholderia cepacia TM1, which was isolated previously, also grew on the above-mentioned compounds because they functioned as the sole carbon source under neutral conditions. Purified VDHs showed activities toward some aromatic aldehydes. These enzymes have the same subunit molecular mass of about 57 kDa as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, but differed in some of their observed properties. Native molecular masses also differed between the purified enzymes. These were 250 kDa for the enzyme from alkaliphilic strain TA1 and 110 kDa for that from neutrophilic strain TM1, as determined by gel filtration. The enzyme from strain TA1 required NADP(+) as a coenzyme for its activity, but that from strain TM1 required NAD(+). These results are important because this is the first report of an alkaliphilic bacterium consuming lignin monomers.
Mitsui
Ryoji
R
Department of Biochemistry, Faculty of Science, Okayama University of Science, Kita-ku, Okayama, Japan. rmitsui@dbc.ous.ac.jp
Hirota
Mizuho
M
Tsuno
Takuo
T
Tanaka
Mitsuo
M
D000445:Aldehyde Oxidoreductases / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D001547:Benzaldehydes / Q000378:metabolism*; D016956:Burkholderia cepacia / Q000201:enzymology* / Q000254:growth & development / Q000378:metabolism; D002850:Chromatography, Gel; D003067:Coenzymes / Q000378:metabolism; D003373:Coumaric Acids / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D004795:Enzyme Stability; D006863:Hydrogen-Ion Concentration; D062385:Hydroxybenzoates / Q000378:metabolism; D008837:Micrococcus / Q000201:enzymology* / Q000254:growth & development / Q000378:metabolism; D008970:Molecular Weight; D009243:NAD / Q000378:metabolism; D009249:NADP / Q000378:metabolism; D013696:Temperature; D014641:Vanillic Acid / Q000378:metabolism
D001547:Benzaldehydes; D003067:Coenzymes; D003373:Coumaric Acids; D062385:Hydroxybenzoates; D009243:NAD; C009091:protocatechuic acid; D009249:NADP; C004999:ferulic acid; C100058:vanillin; D000445:Aldehyde Oxidoreductases; C105452:vanillin dehydrogenase; D014641:Vanillic Acid