PROTOPLASTS FROM PECTINOLYTIC FUNGI - ISOLATION, REGENERATION AND PECTINOLYTIC ENZYME-PRODUCTION

Protoplast release in pectinolytic strain mutants of Aspergillus sp. C H-Y-1043 (A13) and Aspergillus flavipes ATCC-16795 (F7) is described. Optimum yield of protoplasts A13 was obtained in a lapse of 1 h when c ommercially lytic enzymes of Trichoderma harzanium (2 mg ml(-1)) were added in 0.05 mol l(-1) citrate-phosphate buffer pH 5.0 containing 0.7 mol l(-1) KCl and 10 mg ml(-1) BSA. Best results in F7 were obtained when the protoplasting system of A13 was supplemented with 10 mg ml(-1 ) Aureobasidium sp. lytic enzymes. Isolated protoplasts in A13 and F7 were capable of a high regeneration frequency of 87% and 53% when 0.7 mol l(-1) KCl and sorbitol were used as osmotic stabilizers, Endo-P, E xo-P and pectin lyase production were not modified during the process of regeneration.