Antibacterial Activity of a Chitooligosaccharide Mixture Prepared by Cellulase Digestion of Shrimp Chitosan and Its Application to Milk Preservation

Abstract
The antibacterial activity of a chitooligosaccharide mixture prepared by digestion of shrimp chitosan with cellulase at 50°C for 14 h was evaluated. Sugars with 1 to 8 degrees of polymer (DP) were found in this chitooligosaccharide mixture, and the weight percentage of sugars with DP ≧ 6 was 44.3%. Minimal lethal concentrations of this mixture against Aeromonas hydrophila, Escherichia coli, Listeria monocytogenes, Pseudomonas aeruginosa, Salmonella Typhimurium, Shigella dysenteriae, Staphylococcus aureus, Vibrio cholerae, and Vibrio parahaemolyticus in nutrient broth were 5 to 29 ppm, which were much lower than those of the chitosan reactant (50 to 1,000 ppm). The antibacterial activity of this mixture in the sterilized milk against E. coli O157, L. monocytogenes, Salmonella Typhimurium, and S. aureus was much stronger at 4°C than at 37°C. When raw milk was supplemented with either 0.24% or 0.48% (wt/vol) of this oligosaccharide mixture and stored at 4°C for 12 days, its mesophilic and psychrotrophic counts were reduced by at least 3 log cycles, and there was very little change in pH. In addition, this mixture retarded the growth of Salmonella species and caused quicker reduction of Staphylococcus species in raw milk. Accordingly, the shelf life of raw milk at 4°C was extended by at least 4 days.