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United States Department of Agriculture

Agricultural Research Service

Research Project: IMPROVED KNOWLEDGE OF VIRULENCE FACTORS TO DEVELOP POSTHARVEST DECAY CONTROL STRATEGIES Title: Application of the 2-cyanoacetamide method for spectrophotometric assay of cellulase enzyme activity

Authors
item Jurick, Wayne
item Vico, Ivana -
item Whitaker, Bruce
item Gaskins, Verneta
item Janisiewicz, Wojciech

Submitted to: Plant Pathology Journal
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: March 26, 2012
Publication Date: April 17, 2012
Citation: Jurick Ii, W.M., Vico, I., Whitaker, B.D., Gaskins, V.L., Janisiewicz, W.J. 2012. Application of the 2-cyanoacetamide method for spectrophotometric assay of cellulase enzyme activity. Plant Pathology Journal. DOI:10.3923/ppj.2012.

Interpretive Summary: Cellulose is the main component of plant cell walls. Cellulases are enzymes that break down cellulose into glucose which can then be used as an energy source by microorganisms. Cellulases are important enzymes that function in many biological processes in plants and microbes, which require a safe way to measure their activity. The current method used to measure cellulase activity involves the use of carcinogenic and corrosive chemicals. We have developed a new method to accurately measure cellulase activity without the use of such toxic chemicals which performs well. This assay can be used by scientists to measure cellulase enzyme activity in a safer manner.

Technical Abstract: Cellulose is the most abundant form of carbon on the planet. Breakdown of cellulose microfibrils in the plant cell wall is a means by which microbes gain ingress into their respective hosts. Cellulose degradation is also important for global carbon recycling and is the primary substrate for production of biofuels. In this study, we developed a cellulase assay method that rivals the commonly used dinitrosalicylic acid (DNS) assay. It was shown that the 2-cyanoacetamide method is capable of detecting D-glucose in a linear fashion, can function in various buffers at pH ranging from 4.0 to 8.0, and is as sensitive as the DNS test at detecting fungal cellulase activity using carboxymethylcellulose as a substrate. This method will be useful to others who desire to accurately and efficiently assay microbial cellulase activity without the use of phenol and other highly toxic and corrosive chemicals.

Last Modified: 7/31/2014