i am in need to quantify exogluconase production by the bacterial isolates. for this avicel acts as substrate which is insoluble in water and solvent, how to dissolve it and use it for assay?
Hello Sujatha
There is no way to dissolve Avicel. However, you can assay the soluble reducing sugars that exoglucanases will release into the incubation medium.
Agreed, Avicel won't solubilize, it needs to be centrifuged out after enzyme assay has been stopped in order to do spec analysis. Lots of people use DNS for reducing sugars but there are a lot of kits out there for glucose measurement as well.
Just a word of caution regarding Gregory's answer : most exoglucanases are in fact cellobiohydrolases generating cellobiose and not glucose. Cellobiose will not react with glucose reagents based on glucose oxidase (e.g. glucostat), unless cellobiose is hydrolyzed to glucose by beta-glucosidase
You can prepare a suspension of Avicel in water. For taking samples for each testing tube, keep the suspension on agitation, hence you'll have a homogeneous distribution of the Avicel. Take samples with automatic positive displacement pipette.
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