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Amperometric method for the determination of cellulase activity and its optimization using response surface method




Kättesaadav alates


Nero, Georg
Kivirand, Kairi
Othman, Sana Ben
Rinken, Toonika

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Cellulases are a group of enzymes, which catalyse different steps of cellulose hydrolysis, and are broadly used in industry as unpurified mixtures of several enzymes. The total activity of cellulase is defined as the ability of the enzyme to produce glucose, which is the final product of cellulose hydrolysis, and is expressed in cellulase units. However, common strategies for the determination of the cellulolytic activity of industrial cellulase preparations are based on the assessment of different steps of cellulose hydrolysis, and the results obtained with different methods are not similar. The aim of the present study was to develop an assay for the determination of cellulase activity that relies on the amperometric determination of the final product of cellulose hydrolysis glucose. The assay conditions were optimized using response surface methodology (RSM) combined with Box-Behnken design. The detection limit of the proposed method was 1.71 ± 0.06 U. We compared the results of the amperometric method with the ones obtained with the spectrophotometric method and viscosimetry in a commercial cellulose preparation.
This research was funded by the European Union’s Horizon 2020 research and innovation program project VALORTECH under Grant agreement No. 810630 and the Mobilitas Pluss ERA Chair support (No. MOBEC006).



cellulase activity, amperometric detection, response surface method, optimization of assay conditions, articles