Characterization of the optimal conditions for the cultivation of Cladosporium cladosporioide (Fres.) de Vries and isolation of the pectinase produced in solid-state conditions

摘要

Because of the increasing use of microorganism-derived enzymes in current markets, different procedures for enzyme isolation and purification are being studied. This study aimed to define the ideal time of cultivation for Cladosporium cladosporioide (Fres.) de Vries and partially purify the pectinases produced. The activity of pectin methylesterase (PME) and polygalacturonase (PG) was evaluated after a period of 10, 15 and 20 days post-inoculation. Extraction of the enzyme complex was carried out via homogenization in buffer solutions at different pHs (4, 5 and 6) followed by centrifugation and precipitation with ammonium sulfate saturation at concentrations of 20, 40 and 60%. According to our results, a cultivation time of 10 days resulted in the highest activities of both polygalacturonase 105.52 (U/g min) and pectin methylesterase 1480 (U/g min). The results also showed that the benzoate buffer (pH 4.0) constituted the best condition for extraction of both enzymes. The best extraction results were observed for 60%, ammonium sulfate, with a rate of purification was 14.24 for PME and 1.39 for (PG).