Peroxidase (donor: H2O2 oxidoreductase [EC 1.11.1.7]) was purified from the culture broth of the hyphomycete Arthromyces ramosus in the early log phase to show a single band on SDS-PAGE. The crystals of A. ramosus peroxidase (ARP) were formed by salting out with ammonium sulfate at room temperature and pH 7.5. The repeated seeding technique was employed to grow the crystals to the size large enough for X-ray diffraction study. The crystals were characterized as tetragonal, space group P4(2)2(1)2, with unit cell dimensions of a = b = 74.5 A, c = 117.6 A. The asymmetric unit contains one molecule of peroxidase. They diffract X-rays to at least 2.0 A resolution and are stable to X-rays.

Characteristics of yeast growth and ethanol fermentation were examined in membrane bioreactor using a grape juice. After inoculation, batch fermentation was carried out for 24 h. When yeast growth reached the stationary phase, continuous fermentation was initiated. In continuous fermentation, a linear relationship was observed between cell concentration and dilution rate. In single-vessel membrane bioreactor, the cell concentrations of 18.7 g/l and 76.9 g/l (15 and 60 times higher than that of the batch fermentation, respectively) were observed at dilution rates of 0.1 h(-1) and 0.3 h(-1), respectively. The residual sugar concentration was higher than 10 g/l at the dilution rate of 0.1 h(-1), 0.2 h(-1) or 0.3 h(-1), therefore the single-vessel membrane bioreactor was not suitable for producing dry wine (sugar concentration: 4 g/l or less). In the double-vessel membrane bioreactor, it is most suitable to set the recycle ratio at 0.15 for keeping the sugar concentration below 4 g/l. The productivity of dry wine in the double-vessel membrane bioreactor was 28 times higher than that in the batch fermentation.

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Carbon and nitrogen sources were investigated for improving peroxidase production by Arthromyces ramosus, a hyperproducer of peroxidase. Glucose as carbon source and a mixture of yeast extract and polypeptone at the ratio of 3 to 5 as nitrogen source in a production medium were shown to give the highest peroxidase activity. During the culture amino acids such as alanine, arginine, methionine, leucine, tyrosine and tryptophan were depleted. Therefore, glucose supplemented nitrogen source fed-batch culture was carried out and a peroxidase activity of 73 U/ml was obtained. This activity was 1.7 times higher than that of glucose fed-batch culture. This indicates that an adequate nitrogen source supply during the culture is effective for improving the peroxidase production by A.ramosus.

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