Detection of β‐glucanase activity on various β‐1,3 and β‐ 1,4‐glucans after native and denaturing polyacrylamide gel electrophoresis

β-Glucanases were detected after polyacrylamide gel electrophoresis under native and denaturing conditions using various β-1, 3- and β-1, 4-glucans, including mixed glucans (laminarin, pachyman, carboxymethyl cellulose, lichenan and barley β-glucan). After electrophoresis and incubation of gels, substrates incorporated into polyacrylamide gels were stained with specific fluorochromes, Sirofluor for β-1, 3 linkages and Calcofluor White M2R for β-1, 4 linkages. Under UV illumination, lysis zones appeared as dark bands against a fluorescent background. Enzymes of bacterial, fungal and plant sources could be revealed sequentially in gles containing mixed β-(1.3) (1,4)-glucans by staining first with sirofluor followed by staining with Calcofluor White M2R. Active profiles were more diverse when substrates were stained with sirofluor. The use of purified sirofluor at pH 11.5 compared with Aniline Blue at pH 8.6 allowed better detection of β-1, 3-glucanase activities. In gels containing laminarin stained with sirofluor, bands exhibiting a more intense fluorescence than the background fluorescence were observed in addition to dark nonfluorescent bands. It is postulated that these two types of β-1, 3-glucanase activities differ by their enzymatic action (partial versus extensive hydrolysis). Analysis of fungal extracts using denaturing gels embedded with various β-glucans displayed lysis bands migrating between 32 and 35 kDa.