Abstract
Soluble dye-labelled substrates were prepared for the specific and rapid screening and the measurement of endo-β-1,4-glucanase and endo-β-1,4-mannanase activities. Soluble carboxy-methylcellulose and locust bean galactomannan were dyed with Remazol Brilliant Blue R. A specific screening procedure was developed to isolate genes coding for cellulolytic and hemicellulolytic enzymes from a Cellulomonas sp. Ce1B7 library. The assay is advantageous for the simple, rapid and direct detection of cellulase and hemicellulase activities in a large number of clones as found in gene libraries without replica plating or blotting techniques. The prepared dye-labelled substrates can also be used for the quantitative measurement of specific endo activities of cellulolytic and hemicellulolytic enzymes because the undigested substrate can be precipitated and the amount of soluble dye, determined spectrophotometrically, is proportional to enzyme activity.
Published Version
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