Extraction of intracellular chitosanolytic and chitinolytic enzymes from filamentous fungi of the genus Mucor

Abstract

Mucor is a genus of fungi belonging to the Mucorales, the most important order of zygospore-forming fungi. Mucor species are attracting growing interest due to their distinctive features, which make them excellent models for genetic studies, and also due to their wide industrial applications. They are a source of lipids, pigments, biosurfactants, and numerous enzymes, including proteases, lipases, cellulases, chitinases, and chitosanases. However, information regarding the isolation of intracellular chitosanolytic and chitinolytic enzymes from different Mucor species remains limited, and species-specific requirements for enzyme extraction have not been sufficiently investigated. In this context, the aim of this study was to develop a method for extracting intracellular proteins produced by Mucor racemosus CH52 and Mucor hiemalis CH80 and to confirm their activity in chitosan and chitin hydrolysis. Detergent lysis (Triton X-100, 0.5% w/v) was found to be the most effective method for isolating these enzymes from M. racemosus CH52 cells. In the case of the second strain, the best results were obtained by freezing and grinding biomass, combined with lysis using detergent (Triton X-100, 0.5% w/v). These methods enabled the production of cell-free extracts with high protein content and chitosanase and chitinase activities. The results demonstrated species-dependent differences in enzyme recovery and indicated the importance of developing tailored extraction strategies for individual Mucor species.