In different enzyme assay protocols like CMCase, pectinase, amylase, using 1 % or 0.5% substrate. In protocols they did not mention how substrate solution should be prepared whether it will be in distilled water or a specified buffer? kindly recommend me what is the correct method for substrate preparation?
In general, that depends on the stability of the substrate, some substrates, like nucleotides, keep better at a specific pH. With pectin, this is unlikely to be a problem, so you can use buffer or water depending on convenience.
Make contact with Garabed Antranikian (also member of ResearchGate). I assume that he can help you. Say Hello to him and kind regards from me (Frank Mayer)
The substrates you mention are carbohydrates. I am not sure about pectin, but for CMC and starch, they can be autoclaved in distilled water to prevent contamination, and concentrated buffer can be added freshly to prepare the desired amount of assay mixture.
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