Inhibitory effect of chondroitin sulfate oligosaccharides on bovine testicular hyaluronidase

•Chondroitin sulfate (ChS) oligosaccharides inhibited hyaluronidase activity.•Transglycosylation by hyaluronidase was also inhibited by the ChS oligosaccharides.•The inhibitions of hydrolysis and transglycosylation were dose dependent.•The inhibition mode by the ChS oligosaccharides on the hydrolysis was competitive.•Nonsulfated oligosaccharides did not inhibit hyaluronidase activity. Hyaluronan and chondroitin sulfates are prominent components of the extracellular matrices of animal tissues; however, their functions in relation to their oligosaccharide structures have not yet been fully elucidated. The oligosaccharides of hyaluronan and chondroitin sulfate were prepared and used to investigate their effects on the hydrolysis and transglycosylation reactions of bovine testicular hyaluronidase when hyaluronan was used as a substrate. Hydrolysis and transglycosylation activities were assessed in independent reaction systems by analyzing the products by HPLC. The hydrolysis and transglycosylation reactions of bovine testicular hyaluronidase were dose-dependently inhibited by chondroitin sulfate oligosaccharides, but not by hyaluronan or chondroitin oligosaccharides. A kinetic analysis of the hydrolysis reaction using hyaluronan octasaccharide revealed that the inhibition mode by chondroitin sulfate oligosaccharides was competitive.