and contained environment with a controlled temperature
and humidity of 30°C; 94% respectively. In tray
number one (un-inoculated plate, pressmud was mixed
with nutrient mix containing sucrose (1%), ammonium
sulphate (0.4%) and peptone (0.2%) dissolved in 25 mL
of tap water. The composition of media constituents in
various trays was depicted below (Table 6). The contents
of the plates were mixed well and kept for solid state fermentation
for a period of 96 h. Humidity (90-95%) was
maintained by manually spraying of sterile distilled water
in every 12 h. Sample (20 g) were collected at every 12 h
andfermented solid was mixed with the 80 ml of extraction
buffer (100 mM Potassium phosphate, 10 mMβ-
mercaptoethanol and 1 mM phenyl methyl sulfonyl fluoride)
and kept for stirring for 30 min at 10°C. Leachate
was filtered with nylon cloth and centrifuged at 6000 rpm
for 15 min at 10°C. Supernatant was used as crude invertase
and went for invertase assay.