Manufacturing technology of low-molecular-weight chitosan based on enzymatic hydrolysis

Abstract

Chitosan is natural high molecular weight polymer of D-glucosamine and N-acetyl - D - glucosamine connected by 1,4 - b - glycoside bond with a molecular mass of 1000 kDa (and above), practical use is difficult because of high viscosity of its aqueous solutions even at low concentrations, and lack of solubility at neutral pH and, consequently, low biological activity. To reduce viscosity, improve the solubility and enhance biological activity of high molecular weight chitosan subjected to depolymerization. Chitosan, like other polysaccharides, is characterized by a hydrolysis reaction, which is due to the presence of glycoside bonds in the molecule that are lable to hydrolyzing agents, for example, aqueous solutions of acids, alkalis, as well as to the effect of some hydrolases. During hydrolysis, glycoside bonds are broken and, as a result, the molecular weight of chitosan decreases. However, these processes are accompanied by the formation of significant amounts of toxic products and require very costly disposal of waste before it is discharged into the environment. Chitin and chitosan are natural biopolymers and their synthesis, modification and degradation are associated with enzymatic transformations. It is the biodegradability to the usual substances for the body that is one of the main advantages of chitosan. It is obvious that the most appropriate method is the enzymatic hydrolysis of chitosan. As enzyme preparations for the degradation of chitin and chitosan, enzyme complexes of various origins are used. These can be enzymes from crab or krill hepatopancreas complexes, as well as pancreatin from the pancreas of cattle. But more often for this purpose, enzymes complexes with chitinolytic activity of microbiological origin are used. In this study, low-molecular-weight chitosan was obtained by enzymatic hydrolysis using the extracellular chitinolytic complex of Streptomyces kurssanovii. The resulting chitosan had a medium-viscosity molecular weight of 25-40 kDa. Carrying out two stages of fractionation (stepwise acidification and separation on membranes) made it possible to obtain chitosan fractions with a narrow distribution by molecular weight.