Preparation and antibacterial effect of chitooligosaccharides monomers with different polymerization degrees from crab shell chitosan by enzymatic hydrolysis

This study aimed to explore the structure and antibacterial properties of chitooligosaccharide monomers with different polymerization degrees and to provide a theoretical basis for inhibiting Salmonella infection. Chitosan was used as a raw material to prepare and separate low-molecular-weight chitooligosaccharides. Chitobiose, chitotriose, and chitotetraose were obtained by gradient elution with cation exchange resin. The molecular weights and acetyl groups of the three monomers were determined by matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) and nuclear magnetic resonance (NMR), respectively. Three chitooligosaccharide monomers were used to explore the antibacterial effect on Salmonella. The results showed that the degree of deacetylation of chitosan was 92.6%, and the enzyme activity of chitosanase was 102.53 U/g. Within 18 h, chitosan was enzymatically hydrolyzed to chitooligosaccharides containing chitobiose, chitotriose, and chitotetraose, which were analyzed by thin-layer chromatography (TLC) and MALDI-TOF. MALD-TOF and TLC showed that the separation of monomers with ion exchange resins was effective, and NMR showed that there was no acetyl group. Chitobiose had a poor inhibitory effect on Salmonella, and chitotriose and chitotetraose had equivalent antibacterial effects.