Cellulase activity during the growth of Achlya bisexualis on glucose, cellulose, and selected polysaccharides
The antheridial strain of the dioecious water mold Achlya bisexualis was grown in chemically defined media using glucose, cellobiose, and selected polysaccharides as carbon sources. Growth and cellulase levels were measured with media containing glucose, cellobiose, and cellulose. Evaluation of cellulase activity in the medium by viscometric and reducing sugar generation assays suggests that cellulase plays a significant role in degrading cellulose for uptake and catabolism by A. bisexualis. Cellulase in glucose-grown cultures exists as a soluble extracellular enzyme complex, while in cellulose-grown cultures much of the enzyme is absorbed to the cellulose. Elution of the cellulose substrate after 96 h growth with NaCl-fortified buffer releases absorbed cellulase in a soluble form. The absorption of cellulase to the substrate and possibly the cell walls of A. bisexualis could account for the rapid loss in dry weight of A. bisexualis during culture on cellulose in a closed system. Scanning electron microscopic (SEM) examination of the walls of A. bisexualis shows disruption in cellulose cultures, which is not evident for glucose-or cellobiose-grown hyphae. Transmission electron microscopic (TEM) photomicrographs show a significant reduction in the wall thickness of cellulose-grown hyphae as compared with glucose-grown samples. This evidence suggests that the enzyme(s) produced during growth on cellulose is (are) capable of binding as an active hydrolase to walls of A. bisexualis or to the cellulosic substrate.