Summary
Membrane-bound aldehyde dehydrogenase (ALDH) was purified from the membrane fraction of an industrial-vinegar-producing strain, Acetobacter polyoxogenes sp. nov. NBI1028 by solubilization with Triton X-100 and sodium N-lauroyl sarcosinate and subsequent column chromatography on DEAE-Sepharose CL-6B and hydroxyapatite. The purified enzyme was homogeneos on polyacrylamide disc gel electrophoresis. Upon sodium dodecyl sulphate-polyacrylamide gelelectrophoresis, the enzyme showed the presence of two subunits with a molecular mass of 75 000 daltons and 19 000 daltons, respectively. From the absorption and fluorescence spectra, the absence of cytochrome c and the presence of pyrroloquinoline quinone in the purified enzyme were demonstrated. The ALDH preferentially oxidized aliphatic aldehyde with a straight carbon chain except for formaldehyde. The apparent K m for acetaldehyde was 12 mM. The optimum pH and temperature were 7.0 and 50°–60°C, respectively. The enzyme remained active after storage at 4°C for 20 days. p-Chloromercuribenzoic acid and heavy metal salts such as CuSO4 were inhibitory to the enzyme. Ferricyanide was effective as an electron acceptor.
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Fukaya, M., Tayama, K., Okumura, H. et al. Purification and characterization of membrane-bound aldehyde dehydrogenase from Acetobacter polyoxogenes sp. nov.. Appl Microbiol Biotechnol 32, 176–180 (1989). https://doi.org/10.1007/BF00165884
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DOI: https://doi.org/10.1007/BF00165884