How to assay enzyme cellulase?

Question

this is sort of my thesis question. i need to find a way to identify and isolate organism from surround that can synthesis that enzyme... give me any clue please..

Answer 1

Cellulase cleaves cellulose and produce glocose.

Generally for assay 1% soluble cellulose [and not microcrystalline] is used.

Enzyme and substrate are mixed and incubated at proper temp. & pH for 10 - 30 min.

Glucose has chemical reducing capacity, hence it reacts with the reagent DNSA and its colour changes from yellow to red.

In a particular range of glucose conc. absorbance of red colour developed is proportional to glucose conc.

Hence from the standard graph of glucose conc Vs OD in colour reaction cellulase activity can be assayed.

Cellulase is Lytic enzyme of group 6 which has nothing to do with NAD/NADP coenzymes as stated in some other answer.

Answer 2

Cellulase activity is determined by its effect on microcrystalline cellulose with respect to glucose formation. Released glucose is determined in a hexokinase/glucose-6-phosphate dehydrogenase system at 340 nm.

Cellulose -->glucose---> glucose-6-phosphate--->reduced & NADH is produced from NAD. Amount of NADH is measured at 340nm.

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