Cellobiohydrolases Produce Different Oligosaccharides from Chitosan

Area of Science:

• Biotechnology
• Biochemistry
• Enzymology

Background:

• Chito-oligosaccharides (COSs) are valuable bioactive molecules, but their limited availability hinders widespread application.
• Current production methods face challenges in scalability and accessibility of starting materials.

Purpose of the Study:

• To explore the potential of readily available glycosidases, specifically cellobiohydrolases, for the efficient production of chito-oligosaccharides (COSs).
• To compare the efficacy of cellobiohydrolases with traditional chitosanases in generating COSs with defined characteristics.
• To elucidate the enzymatic cleavage mechanisms and structural variations in produced COSs.

Main Methods:

• Hydrolysis of chitosan using selected glycosidases, including cellobiohydrolases (EC 3.2.1.91) and chitosanases (EC 3.2.1.132).
• Analysis of produced oligosaccharide fractions using Mass Spectrometry (MS) and Nuclear Magnetic Resonance (NMR) spectroscopy.
• Detailed structural characterization, including degree of polymerization and degree of acetylation (DA).

Main Results:

• Cellobiohydrolases demonstrated significant potential for COS production, yielding oligosaccharides with a degree of polymerization between three and six units.
• The degree of acetylation (DA) of the produced COSs varied depending on the specific enzyme employed.
• NMR analysis identified preferred enzymatic cleavage sites, revealing distinct chitosanase and chitinase activities of the tested cellobiohydrolases.

Conclusions:

• Readily available cellulolytic enzymes offer a viable alternative to chitosanases for producing chito-oligosaccharides (COSs).
• This enzymatic approach allows for the generation of COSs with distinct structural features, enhancing accessibility to this important class of bioactive compounds.
• The findings support the broader exploitation of enzymatic hydrolysis for tailored COS production.