All of the freshly inoculated flasks
were incubated at 30 1 C in a shaking incubator at 120 rpm
for 5 days of fermentation period. After every 24 h the crude
pectin lyase was extracted by mixing the fermented material
with 100 mL of 100 mM citrate buffer followed by keeping
them in continuous shaking position (120 rpm) for half an
hour. The biomass was then filtered by passing through
125 mm filter paper followed by centrifugation at 3000 g for
10 min at 4 C. The resulting supernatants were used as crude
extract for enzyme activity estimation. The weight of biomass
was determined in pre weighted petri plates and kept in oven
for 24 h at 100 C, till constant weight is obtained.