Auxotrophic strains of Aspergillus niger were obtained from citric-acid-producing strains of the fungus after irradiation with UV light. Protoplasts were isolated from young hyphae of the auxotrophic strains after treatment with snail enzyme and than treated with polyethylene glycol (30%, W/V), in a Ca2+ (10 mmol/L) solution. The pH value of the suspension was adjusted to 9.0. The frequency of the heterokaryons (related to the number of protoplasts reverting after PEG treatment) was 0.67%. Prototrophic heterozygous spores were isolated from a heterokaryon with the frequency of 1.2 x 10(-6). Citric acid production in the best heterozygous strains was about 15% higher than that of the high-production parent strain.
A considerable lowering of aeration demands occurs during diffuse growth of citric acid-producing Aspergillus niger in a submerged cultivation. However, the diffuse culture poses stricter demands on the type of aeration and agitation. The impeller frequency affects considerably the morphology of the producer fungus and the accumulation of citric acid. The effect of impeller frequency on the distribution of air in the medium and on the amount of air fed into the diffuse culture is less important.
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