Purification and properties of an aryl-alcohol dehydrogenase from the white-rot fungus Phanerochaete chrysosporium.
An intracellular aryl-alcohol dehydrogenase (previously referred to as aryl-aldehyde reductase) was purified from the white-rot fungus Phanerochaete chrysosporium. The enzyme reduced veratraldehyde to veratryl alcohol using NADPH as a cofactor. Other aromatic benzaldehydes were also reduced, but not aromatic ketones. Methoxy-substituted rings were better substrates than hydroxylated ones. The enzyme was also able to reduce a dimeric aldehyde (4-benzyloxy-3-methoxybenzaldehyde). The highest reduction rate was measured when 3,5-dimethoxybenzaldehyde was used as a substrate. On SDS/PAGE the purified enzyme showed one major band with a molecular mass of 47 kDa, whereas gel filtration suggested a molecular mass of 280 kDa. Polyclonal antibodies raised against the gel purified 47-kDa protein were able to immunoprecipitate the aryl-alcohol dehydrogenase indicating that its activity possibly resides entirely in this protein fragment. The pI of the enzyme was 5.2 and it was most active at pH 6.1. The aryl-alcohol dehydrogenase was partially inhibited by typical oxidoreductase inhibitors.
Muheim
A
A
Department of Biotechnology, Eidgenössische Technische Hochschule-Hönggerberg, Zürich, Switzerland.
Waldner
R
R
Sanglard
D
D
Reiser
J
J
Schoemaker
H E
HE
Leisola
M S
MS
D000429:Alcohol Oxidoreductases / Q000737:chemistry* / Q000302:isolation & purification; D000447:Aldehydes / Q000378:metabolism; D001592:Benzyl Alcohols / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D004795:Enzyme Stability; D005658:Fungi / Q000187:drug effects / Q000201:enzymology*; D006863:Hydrogen-Ion Concentration; D008031:Lignin / Q000378:metabolism; D008970:Molecular Weight; D009249:NADP / Q000378:metabolism; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D013379:Substrate Specificity
D000447:Aldehydes; D001592:Benzyl Alcohols; D011500:Protein Synthesis Inhibitors; D009249:NADP; D008031:Lignin; D000429:Alcohol Oxidoreductases; C055882:aryl-alcohol dehydrogenase; C042197:veratryl alcohol