Enzymatic Synthesis and Characterization of Different Families of Chitooligosaccharides and Their Bioactive Properties

Chitooligosaccharides (COS) are homo- or hetero-oligomers of D-glucosamine (GlcN) and N-acetyl-D-glucosamine (GlcNAc) that can be obtained by chitosan or chitin hydrolysis. Their enzymatic production is preferred over other methodologies (physical, chemical, etc.) due to the mild conditions required, the fewer amounts of waste and its efficiency to control product composition. By properly selecting the enzyme (chitinase, chitosanase or nonspecific enzymes) and the substrate properties (degree of deacetylation, molecular weight, etc.), it is possible to direct the synthesis towards any of the three COS types: fully acetylated (faCOS), partially acetylated (paCOS) and fully deacetylated (fdCOS). In this article, we review the main strategies to steer the COS production towards a specific group. The chemical characterization of COS by advanced techniques, e.g., high-performance anion-exchange chromatography with pulsed amperometric detection (HPAEC-PAD) and MALDI-TOF mass spectrometry, is critical for structure-function studies. The scaling of processes to synthesize specific COS mixtures is difficult due to the low solubility of chitin/chitosan, the heterogeneity of the reaction mixtures, and high amounts of salts. Enzyme immobilization can help to minimize such hurdles. The main bioactive properties of COS are herein reviewed. Finally, the anti-inflammatory activity of three COS mixtures was assayed in murine macrophages after stimulation with lipopolysaccharides.

Automated summary

Chitooligosaccharides (COS) are oligomers of D-glucosamine and N-acetyl-D-glucosamine obtained by chitosan or chitin hydrolysis. Enzymatic production is preferred for its mild conditions and efficiency in controlling product composition. Selecting the right enzyme and substrate properties is crucial in directing synthesis towards specific COS types, but scaling processes for specific mixtures is challenging due to low solubility and reaction heterogeneity. Immobilizing enzymes can help overcome these hurdles, and COS have bioactive properties including anti-inflammatory activity.