Microplate-based filter paper assay to measure total cellulase activity.
The standard filter paper assay (FPA) published by the International Union of Pure and Applied Chemistry (IUPAC) is widely used to determine total cellulase activity. However, the IUPAC method is not suitable for the parallel analyses of large sample numbers. We describe here a microplate-based method for assaying large sample numbers. To achieve this, we reduced the enzymatic reaction volume to 60 microl from the 1.5 ml used in the IUPAC method. The modified 60-microl format FPA can be carried out in 96-well assay plates. Statistical analyses showed that the cellulase activities of commercial cellulases from Trichoderma reesei and Aspergillus species determined with our 60-microl format FPA were not significantly different from the activities measured with the standard FPA. Our results also indicate that the 60-microl format FPA is quantitative and highly reproducible. Moreover, the addition of excess beta-glucosidase increased the sensitivity of the assay by up to 60%.
Centre for Structural and Functional Genomics, Concordia University, 7141 Sherbrooke Street West, Montreal H4B 1R6 Quebec, Canada.
Zhizhuang
Z
Xiao
Reginald
R
Storms
Adrian
A
Tsang
D002480:Cellulase / Q000032:analysis* / Q000737:chemistry*; D002623:Chemistry Techniques, Analytical / Q000295:instrumentation* / Q000379:methods* / Q000592:standards; D004789:Enzyme Activation; D005947:Glucose / Q000737:chemistry*; D008851:Micropore Filters; D010209:Paper; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D002480:Cellulase; D005947:Glucose