Abstract: Endo-β-N-acetylglucosaminidase H (Endo H) is a glycohydrolase which cleaves the β-1,4-glycosidic bonds of the N-acetylglucosamine core of oligosaccharides and leaves one N-acetylchitobiose attached to the asparagine residue of the glycoprotein. It can be used to identify the glycosylation sites. In this study, we cloned the endoH gene from the Streptomyces avermitilis, then subcloned into expression vector pMA0911.1. We expressed EndoH in Bacillus subtilis WB600. The fermentation broth was purified by hydrophobic chromatography and ionic exchange, and relatively purified EndoH was obtained. Utility of EndoH in digestion of RNaseB and identification of N-glycan structure in OVA indicated the potential of recombinant EndoH in research of N-glycan and glycomic.

Key words: endo-β-N-acetyglucosaminidase H(EndoH), RNaseB, the chicken ovalbumin standard glycoprotein (OVA), N-Glycan