2.4 Production of Pectinase
The inocula were standardized to optical density 0.3 (Demirkan, 2011) and were inoculated at 1% (v/v)into the defined enzyme production medium containing 1 g citrus pectin, 0.2 g (NH4)2SO4, 0.6 g
K2HPO4, 0.2 g KH2PO4 and 0.01g MgSO4.7H2O in 100 ml of distilled water. The initial pH was adjusted
to 8.0 and sterilized under pressure at 121°C for 20 min. The medium inoculated in duplicates were
incubated at 35ºC for 48 h in an orbital shaker incubator at 150 rpm. Thereafter, the cultures were
centrifuged at 12000 rpm for 20 min and the cell free supernatants were used to evaluate pectinase activity (Kumar and Sharma, 2012).