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Application of the 2-Cyanoacetamide Method for Spectrophotometric Assay of Cellulase Enzyme Activity

Jurick II, W. M., Vico, I., Whitaker, B. D., Gaskins, V. L., Janisiewicz, W. J.
Plant Pathology Journal 2012 v.11 no.1 pp. 48
assays, buffers, carbon, carboxymethylcellulose, cell walls, cellulose, endo-1,4-beta-glucanase, enzyme activity, enzyme substrates, fungi, glucose, hosts, pH, phenol, recycling, salicylic acids, toxic substances
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.