Abstract
Differences in polysaccharide composition of various fungal cell walls were indicated by their susceptibility to enzymatic digestion. This information was used to optimize the enzymatic extraction of intracellular enzymes or the preparation of fungal protoplasts in high yield. Bacterial glucanase and chitinase specially purified were used for this study. Mycelium of Aspergillus niger grown on uric acid was treated with mixtures of glucanase and chitinase. Cell wall breakdown products were analysed and the ratio of chitin to glucan was estimated to be 1:1.4. A. niger protoplast formation was optimized using this information. When the mixture of chitinase to glucanase was 1:1.4, similar to the fungal cell wall composition, a 95% yield of protoplasts was obtained after 30 min and their mean size was 7 μm. However, a ratio of 1.5 to 1 (chitinase to glucanase) was needed for the maximum extraction of uricase. Yield was 10.5 μ g −1 cells after 1.5 h incubation at 28°C. Glucanase alone resulted in a maximum yield of 1.9 μ g −1 while chitinase alone yielded 6.0 μ g −1 under the same conditions.
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