Preparation of seed culture
Vegetative inoculums were used in the present studies.
Fifty millilitre of inoculums medium containing nutrient
broth 13 g/l, pH 7.4 ± 0.2 was transferred to a 100 ml
conical flask and cotton plugged. It was sterilized in an
autoclave at 15 lbs/inch2 pressure at 121°C for 20 min.
After cooling to room temperature, a loopful of freshly
grown culture was aseptically transferred to it. The flask
was incubated overnight at 37°C and 150 rpm in a rotary
shaking incubator.
Enzyme