Cellulase enzyme sold in U/mg, to determine the concentration of protein in this enzyme, what should be done. Is it possible to get 1g/l of protein by dissolving 1 gram of enzyme in one liter of water (considering enzyme is only protein) or Bradford experiment shall be done? Thanks
If the cellulase preparation is pure cellulase without anything else, then you can dissolve 1 gram in 1 liter to make 1 gram enzyme/liter. However, in many cases dry enzymes contain excipients, such as buffers, salt and sugars. In that case, you should use a protein assay on a test solution to find out how much of the mass is protein. Another consideration is that the protein may not be pure cellulase, but it may be contaminated with other proteins. You can run a sample on SDS-PAGE to find out.
Whenever one talks about cellulase, it is not a single enzyme, instead of a cocktail of at least three enzymes, i.e. endoglucanase, cellobiohydrolase, and endoglucanase. Most commercial preparation of cellulase is a crude extract from lysate either in liquid form or solid powder. That's why commercial preparation has activity in unit/mg. You can verify your cellulase mix in SDS PAGE. Quantify total protein using Bradford and then run 10 to 30 ug per lane. You can see the purity of the cellulase mix.
If you let me know what cellulase mix you are using, I will be able to write more precisely.
Thank you both Adam B Shapiro and Sushant Kumar Sinha. Sushant Kumar Sinha the enzyme is going to be used for hydrolysis of cellulose (microcrystalline powder) to produce glucose. The enzyme is lyophilized form from sigma sold in "KU". It has so many species of source, no idea which one to choose. Additionally, most previously done research papers use "FPU" , no idea also equivalent value of "KU" to "FPU" Thanks
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