The production profile of cellulases of the mutant strain A19 from the filamentous fungus Stachybotrys microspora was studied in the presence of various carbon sources (glucose, lactose, cellulose, carboxymethylcellulose (CMC), and wheat bran) and a range of medium initial pH (5, 7, and 8). Two extracellular cellulases from the Stachybotrys strain (endoglucanases and β-glucosidases) were monitored by enzymatic assay, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and zymogram analysis. Glucose and lactose repressed CMCase time course production while they permitted a strong β-glucosidase one. On Avicel cellulose, CMC, and wheat bran, both activities were highly produced. Wheat bran (WB) is the best carbon source with an optimum of production at days 5 and 6. The production kinetics of both activities were shown to depend on the medium initial pH, with a preference for neutral or alkaline pH in the majority of conditions. The exception concerned the β-glucosidase which was much more produced at acidic pH, on glucose and cellulose. Most interestingly, a constitutive and conditional expression of an alkaline endoglucanase was revealed on the glucose-based medium at an initial pH of 8 units. The zymogram analysis confirmed such conclusions and highlighted that carbon sources and the pH of the culture medium directed a differential induction of various endoglucanases and β-glucosidases.
Neutral and alkaline cellulases from microorganisms constitute a major group of the industrial enzymes and find applications in various industries. Screening is the important ways to get novel cellulases. Most fungal cellulases have acidic pH optima, except some fungi like Humicola insolens species. However, new applications require the use of neutral and alkaline cellulases in food, brewery and wine, animal feed, textile and laundry, pulp and paper industries, agriculture as well in scientific research purposes. Indeed, the demand for these enzymes is growing more rapidly than ever before, and becomes the driving force for research on engineering the cellulolytic enzymes. Here, we present an overview of the biotechnological research for neutral and alkaline cellulases.
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