Isolation, Identification and Characterization of Pectinase Producers from Agro Wastes (Citrus sinensis and Ananas comosus)

Fruits detached from the living plants cannot repair its cells or defend its tissues against natural deterioration. Pectinases, the first enzymes to be secreted by fungal pathogens, weakens the plant cell wall and expose other polymers to degradation by hemicellulases and cellulases, are of significant importance in current biotechnological era, with their applications in various industries. The objective of this study, is to isolate and characterize pectinase producers from Bacillus subtilis and Aspergilus niger based on their carbon and nitrogen sources, using different assays. Fresh and rotten orange and pineapple peel were incubated in nutrient broth and the resulting microorganisms Bacillus subtilis and Aspergilus niger were subculture in pectin agar. Pectinase was extracted with pectin agar incubated at room temperature for 6 days. Clear zones of pectinase were isolated for secondary screening by estimating polygalacturonase (PG) and pectin lyase (PL) production under solid state fermentation by using the modified medium. Pectinase activity was analysed by alternating certain parameter, such as temperature and pH. Concentration of D-galacturonic acid and polygalacturonase were adjusted to investigate pectinase activity. The effect of inoculum size (106 CFU/ml, 105 CFU/ml, 104 CFU/ml and 103 CFU/ ml) for the production of pectinase. From this study, 104 CFU/ml and 103 CFU/ml were optimum inoculum sizes for the production of pectinase for both rotten and fresh oranges and pineapples respectively with level of significance (p < 0.05). Optimum pectinase activity was recorded at pH 5 and temperature 35 °C and 30 °C and pH5 and temperature 30 °C and 35 °C for both fresh and rotten oranges and pineapples respectively. Considering its great significance, with tremendous potential pectinase offers industries, one can conclude that strategic ways to produce maximum pectinase with low cost production should be studied