PRODUCTION, CHARACTERIZATION, APPLICATION OF CHITOLOGIOSACCHARIDES HYDROLYSATE WITH PARTIALLY PURIFIED CHITOSANASE ENZYME FROM MARINE ISOLATE BREVIDOMONAS DIMINUTA

Microbial chitosanase has received greater attention for producing chitooligosaccharides. In this study, chitooligosaccharide hydrolysate was developed using the chitosanase enzyme, and its bioactivity was evaluated. Marine muddy samples proved to be a source of isolating novel chitosan-degrading bacteria when compared to the other samples collected. A chitosan-degrading marine bacterium was isolated, and the characterization of its extracellular enzyme, chitosanase, was studied. The molecular characterization and 16S rDNA sequence evolutionary relatedness of the isolate were carried out. The organism was identified as Brevidomonas diminuta, and its sequence was deposited. The production of chitosanase enzyme was significantly induced by the chitosan substrate, while it reached its peak after 72 hours. pH and temperature have been verified to be ideal for enzyme synthesis at 6.5 and 30 °C, respectively. Utilizing a culture medium containing xylose as the substrate at a level ranging from 1.0 to 1.5% significantly enhanced the enzyme production. The molecular weight of the enzyme, 43 KDa, was ascertained through SDS-PAGE. The pathogen’s growth was suppressed by the chitooligosaccharide hydrolysate enzyme. It has promising antimicrobial properties, exhibiting a MIC₅₀ of 0.2 µg/ml against the test pathogens.