RESUMO
The formation of (1-4)-, (1-3)- and (1-6)-beta-glucanases and beta-glucosidases was studied during the growth of the fungus Geotrichum candidum under the conditions of submerged cultivation in a medium optimal for the production of cellulolytic enzymes. Endo-(1-4)-beta-glucanases and C1 enzyme, as well as (1-3)- and (1-6)-beta-glucanases appeared in the medium as soon as by the 45th hour of growth. However, the maximal concentration of the enzymes in the medium was observed at different periods of the fermentation: between 75th and 105th, 70th and 95th, 55th and 100th, 80th and 105th hours, respectively. The content of the enzymes abruptly decreased by the 160th hour of the growth. The activity of beta-glucosidases, which was low at the beginning of the growth, sharply increased by the 70th hour and remained at the same level by the 160th hour of the growth. The accumulation of beta-glucanases was an uneven process, consistent with irregular changes in the content of DNA and protein in the biomass. The isoelectric points of beta-glucanases and beta-glucosidases were studied in the filtrate of the cultural broth after 96 h of the cultivation. The high activity of endo-(1-4)-beta-glucanase was found at the pH 4.6, 4.1 and 3.8; its low activity was detected at the pH 6.4, 3.2, 1.6 and 1.3. Other glucanases behaved also as acid proteins. During isoelectric focusing, (1-3)-beta-glucanase showed the peaks of activity at the pH 4.4, 4.0, 3.8 and 2.9; (1-6)-beta-glucanase, at the pH 5.0, 3.7, 3.5, 3.1 and 2.0; beta-glucosidases were distributed over a broad pH range from 6.7 to 2.0, with the maximal activity at the pH 6.2, 4.8 and 3.7.
