Adney William S, Dowe Nancy, Jennings Edward W, Mohagheghi Ali, Yarbrough John, McMillan James D
Research Triangle Institute, International Research Triangle Park, Durham, NC, USA.
Methods Mol Biol. 2012;908:169-80. doi: 10.1007/978-1-61779-956-3_16.
Although a poor indicator of how a cellulase preparation will perform on biomass, the filter paper unit (FPU) still finds wide use in the literature as an apparent measure of performance efficacy. In actuality, the assessment of commercial enzyme preparation performance in terms of biomass conversion or solubilization of insoluble polysaccharides is largely dependent on the substrate composition, which cannot be easily standardized. Commercial cellulase preparations are evaluated based upon their performance or specific activity. The ability to compare commercial enzyme preparation efficacy across a wide variety of different preparations requires defining the amount of enzyme protein required in milligrams per gram of cellulose to achieve a targeted level of cellulose hydrolysis in a specified timeframe. Since biomass substrates are highly variable, reproducible and accurate protein determination is as important as performance testing to be able to rank order the effectiveness of diverse preparations. This chapter describes a protocol that overcomes many of the difficulties encountered with determining the protein concentration in commercial cellulase preparations.
