A method for the detection and differentiation of cellulase components in polyacrylamide gels |
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| Authors: | T D Bartley K Murphy-Holland D E Eveleigh |
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| Institution: | Department of Biochemistry and Microbiology, Rutgers University-Cook College, New Jersey Agricultural Experiment Station, New Brunswick, New Jersey 08903 USA |
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| Abstract: | Endoglucanase and exoglucanase components of cellulase can be detected and differentiated after polyacrylamide gel electrophoresis by performing activity stains. Endoglucanase activity was visualized in carboxymethyl cellulose agar replicas of gels by staining with Congo red. General beta-1,4-glucanase activity was located by soaking the gel in a solution of NaBH4-reduced cellulooligosaccharides, and detecting the formation of reducing sugars by reaction with triphenyl tetrazolium chloride. Endoglucanases are active in both assays, while exoglucanases can be distinguished by their activity in the cellulo-oligosaccharide assay only. This methodology has facilitated the purification and characterization of cellulase components from Trichoderma reesei and Microbispora bispora. |
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| Keywords: | cellulase gel electrophoresis proteins Microbispora bispora activity stains cellobiohydrolase |
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