Hi everyone,
I want to perform a cellulase activity assay using industrial cellulase and industrial cellulose (microcrystalline cellulose). In case of cellulase produced by the hydrolytic microorganisms, the crude fermentation broth (centrifuged/filtered) containing cellulase is mixed with the cellulosic substrate in appropriate buffer solution, incubated and then analyzed.
The industrial cellulase comes as a solid substance. So, when this cellulase is to be used for the same assay, should it be prepared as an enzyme solution using the common buffers (acetate, citrate, phosphate)? If so, then is there any protocol for preparing the enzyme solution and using the same for the assay? What are the major affecting factors and their optimum conditions in cellulase activity assay?
Regards,
Shiladitya
Since you work with an industrial cellulase, you should check the instructions of the supplier. Buffer conditions will depend on the source of the enzyme mix and the supplier should know the optimal conditions
There are different types of "industrial cellulases", beta-cellulase alfa cellulase among others. The key is to know exactly which type of cellulase you are working with and then select the substrate. Perphaps CMC is an interesting one for your assays.
Regards,
Ana
Hi Ana,
Thank you for your response. I will use some different substrates for the assay. I will also use CMC, by the way. Actually my question is, how to prepare the enzyme solution using the solid enzyme power, which is a protein. Should I use sterile distilled water or a specific buffer with a particular pH? Does this affect the activity of the enzyme?
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