PRODUCTION AND CHARACTERIZATION OF CHITOOLIGOSACCHARIDE HYDROLYSATE PREPARED FROM CHITOSANASE ENZYME OF MARINE ISOLATES

Abstract

Objective: The present study was carried out to develop an enzymatic hydrolysate with unique biological properties targeting diabetic foot ulcers. Methods: Chitosanase-producing organisms were isolated and used to create chitooligosaccharide hydrolysate. Various techniques, such as FTIR, NMR, and X-ray diffraction, were used. Antimicrobial activity was tested using disc diffusion and well diffusion methods. Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) were determined through the Chitooligosaccharide-Broth Dilution Method. Results: The study identified marine mud samples and isolated S9, S15, and SF12 as significant sources of chitosanase production. The partially purified chitosanolytic enzymes produced by these isolates were hydrolyzed in a 1% chitosan solution at 180 °C, revealing more prominent antimicrobial activity. The Chitooligosaccharide Hydrolysate (COS) preparation was fixed at 45 °C, pH 5.5, for 180 min. The chitosanase enzyme was soluble in four solvents and insoluble in ethanol, acetone, and diethyl ether. All COS hydrolysates prepared showed antimicrobial activity against foot ulcer pathogens, Pseudomonas sp., and Candida albicans. S9 COS showed higher activity than SF12 hydrolysates against foot ulcer pathogens. The COS hydrolysate showed significantly stronger antimicrobial activities than chitosan and chitosanase. Conclusion: The present study concludes that COS hydrolysate and its biological functions are applicable for diabetic foot ulcer treatment. Further investigation into the efficacy of COS against diverse infectious pathogens is needed.