Estimation of Cellulase enzyme activity
l.0 ml 0.05 M Na-citrate(pH 4.8) is added to a test tube of volume at least 25 ml.0.5 ml enzyme is diluted in citrate buffer with Whatman No. l filter paper strip(l.0 6.0 cm or 50 mg). Two dilutions are made of each enzyme sample for investigation. One dilution should release slightly more and one slightly less than 2.0 mg(absolute amount) of glucose reducing sugars as glucose) in the reaction conditions. Enzyme solution containing filter paper strip is incubated 50°C for 60 min Then 3.0 ml of DNS reagent is added and mixed well and boiled for exactly 5.0 min in a vigorously boiling water bath. 1ml of 40% Rochelle salt solution is added when the contents of the tubes are still warm. 20ml of double distilled water is added with the content. The intensity of dark red color product is spectrophotometrically read at 510 nm UV/Vis double beam Biospectrophotometer using blank as reference. Similar procedures are followed for the other enzyme concentrations