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Cellulase assays

Cellulase assays

Cellulase enzymes show activity during the ripening of some fruits, where their effects on cell walls results in softening of the fruit. In cases of programmed cell death, such as the formation of aerenchyma (large air spaces in the cortex of plants in flooded soils), and in the abscission zones of leaves and fruits, cellulases are once again very active, breaking down the cellulose walls of the dead cells. Two possible protocols are described here, a gel diffusion assay method and a viscosity reduction assay method.

Gel diffusion method

A fruit extract is placed in a well in some agar containing carboxymethylcellulose (CMC), and, as cellulase molecules diffuse outwards from the well, they destroy cellulose molecules. The more concentrated the enzymes in the well, the larger the zone of cellulose destruction.

Wells containing cellulase should have a clear zone around them, and the diameter of the zone gives a measure of the cellulase activity in that well.

Viscosity reduction method

The technique is based on the action of cellulase enzymes which shorten the lengths of cellulose molecules in a viscous solution of wallpaper paste and cause it to become less viscous (runnier).

The more active the enzyme, the greater the reduction in viscosity, and so the shorter the drainage times.

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