Quantification of Cellulase Activity Using the Quartz Crystal Microbalance Technique

The development of more efficient utilization of biomass has received increased attention in recent years. Cellulases play an important role in processing biomass through advanced biotechnological approaches. Both the development and the application of cellulases require an understanding of the acti...

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Bibliographic Details
Published in:Analytical chemistry (Washington) 2009-03, Vol.81 (5), p.1872-1880
Main Authors: Hu, Gang, Heitmann, John A, Rojas, Orlando J
Format: Article
Language:English
Subjects:
Analytical chemistry
Cells
Cellulase - chemistry
Cellulase - metabolism
Cellulose - metabolism
Chemistry
Chromatographic methods and physical methods associated with chromatography
Crystallization
Crystals
Electrochemistry - methods
Enzymes
Equipment Design
Exact sciences and technology
General, instrumentation
Ion chromatography
Models, Chemical
Nanotechnology
Other chromatographic methods
Quartz - chemistry
Surface Properties
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Summary:The development of more efficient utilization of biomass has received increased attention in recent years. Cellulases play an important role in processing biomass through advanced biotechnological approaches. Both the development and the application of cellulases require an understanding of the activities of these enzymes. A new method to determine the activity of cellulase has been developed using a quartz crystal microbalance (QCM) technique. We compare the results from this technique with those from the IUPAC (International Union of Pure and Applied Chemistry) dinitrosalicylic acid (DNS) standard method and also from biccinchoninic acid and ion chromatography methods. It is shown that the QCM technique provides results closer to those obtained by measuring the actual reducing sugars. The elimination of the use of color development in the standard redox methods makes the QCM platform easier to implement; it also allows more flexibility in terms of the nature of the substrate. Finally, validation of the proposed method was carried out by relating the crystallinity of different substrates to the cellulase activity. Numerical values of cellulase activities measured with the QCM method showed that celluloses with higher crystallinity indices were hydrolyzed slower and to a lower extent than those of lower crystallinity indices for the cellulase mixtures examined.
ISSN:0003-2700
1520-6882
DOI:10.1021/ac802318t