Digestate-Based Liquid Growth Medium for Production of Microbial Chitosan

This study investigated the feasibility of using both the solid and the liquid fractions of waste from the anaerobic digestion process—the digestate—as a possible liquid growth medium for fungal production of chitosan. An enriched liquid phase (ELP), combining both fractions, and derived from mild acid hydrolysis treatment at 120 °C with 6% H2SO4 (w/v) for 70 min, was screened for its ability to support biomass and chitosan production by 17 fungal strains. The best results were obtained with Absidia blakesleeana NRRL 2696 and Rhizopus oryzae NRRL 1510 cultures, which yielded chitosan volumetric productions of 444 and 324 mg L−1, respectively. The chitosan preparations of the former and the latter strain, characterized by infrared spectroscopy, elemental analysis, viscosimetry and thermogravimetric analysis, showed deacetylation degrees of 79% and 84.2%, respectively, and average viscosimetric molecular weights of around 20 and 5.4 kDa, respectively. Moreover, both fungal chitosan samples exerted significant antibacterial activity towards Gram-negative (i.e., Pseudomonas syringae and Escherichia coli) and Gram-positive (i.e., Bacillus subtilis) species.