抄録
For the first time we found that the bacterial strain Gluconobacter oxydans DSM 4025 produced L-ascorbic acid from L-gulono-γ-lactone and therefore we isolated and Purified the enzyme L-gulono-γ-lactone dehydrogenase catalyzing the oxidation reaction. G. oxydans DSM 4025 produced 8.57 and 13.9 mg of L-ascorbic acid Per ml from 70.3 and 89.3 mgof L-gulono-γ-lactone per ml in the growing and resting cell systems, respectively. The enzyme was isolated from the soluble fraction of the cells of G. oxydans DSM 4025 by DEAE-cellulose, Q-Sepharose, hydroxylapatite, and Sephacryl S-300 column chromatographies. The molecular weight of the enzyme was 110, 000 and it consisted of three kinds of subunits of 61, 000, 32, 500, and 16, 5000. L-Gulono-γ-lactone was oxidized to L-ascorbic acid very rapidly in the presence of dyes, such as 2, 6-dichlorophenolindophenol or phenazine methosulfate, but oxygen was not available as an electron acceptor. The absorption spectrum of the reduced form of the native enzyme showed maxima at 416, 521, and 552 nm in the visible region, indicating the presence of cytochrome c. The enzyme isolated from G. oxydans DSM 4025 was entirely different from isofunctional enzymes of eucaryotic organisms.
