Abstract
The versatile oligosaccharide biopolymers, chitin and chitosan, are typically produced using enzymatic processes. However, these processes are usually costly because chitinases and chitosanases are available in limited quantities. Fortunately, a number of commercial enzymes can hydrolyze chitin and chitosan to produce long chain chitin or chitosan oligosaccharides. Here, a platform to screen for enzymes with chitinase and chitosanase activities using a single gel with glycol chitin or glycol chitosan as a substrate was applied. SDS-resistant chitinase and chitosanase activities were observed for pancreatin. Its chitotriosidase had an optimal hydrolysis pH of 4 in the substrate specificity assay. This activity was thermally unstable, but independent of 2-mercaptoethanol. This is the first time a chitotriosidase has been identified in the hog. This finding suggests that oligochitosaccharides can be mass-produced inexpensively using pancreatin.
Highlights
Chitin, an insoluble linear (1→4)-β-linked homopolymer of N-acetyl-D-glucosamine (GlcNAc), is the most abundant polysaccharide in Nature and is distributed ubiquitously
Long-chain oligomers can be obtained from chitin or chitosan that has been digested by enzymes
It has been reported that CBB R250, an acidic dye, can adsorb chitin and chitosan
Summary
An insoluble linear (1→4)-β-linked homopolymer of N-acetyl-D-glucosamine (GlcNAc), is the most abundant polysaccharide in Nature and is distributed ubiquitously. Enzymatic procedures are preferred because chitinases or chitosanases produce more homogeneous oligosaccharides with higher molecular weights These reactions are limited due to the poor stability, low activities and high costs of those enzymes. The first step, which usually occurs in microbes and plays a key role in the solubilization and mineralization of chitin, is the random hydrolysis of the glycosidic bonds between GlcNAc residues by endochitinases to yield chitin oligomers This is followed by further digestion from the nonreducing ends by the monomer-producing exochitinases known as β-N-acetylhexosaminidases. Enzymes with unexpected chitosanolytic and chitinolytic activities have been identified These include glycanases, cellulases, amylases, dextranases, hemicellulases and pectinases. A screening platform was introduced to evaluate enzymes active on chitin or chitosan Using this technique, pancreatin was found to have SDS-resistant (SDSR) chitinase and chitosan activities. Further substrate assays showed that the pancreatin possessed chitotriosidase activity
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