Application

Endo-β-galactosidase was used in fractional protein isolation. It was used for deglycosylation in glycoproteomics of the endothelial secretome of human endothelial cells.

Biochem/physiol Actions

Internal β(1-4) galactose linkages in unbranched, repeating poly-Nacetyllactosamine [GlcNAc β(1-3)Gal β (1-4)] structures are the preferred substrate.Sulfated structures such as keratan sulfate are also cleaved. Branching and/or fucosylation of the substrate may reduce or completely inhibit cleavage. Sulfation of C-6 on galactose will block cleavage. Oligosaccharides of the neolacto-group are cleaved at greatly reduced rates depending on the deviation from the preferred substrate. For example, Gal β(1-3)GlcNAc β(1-3) Gal β(1-4)Glc is cleaved at 5X10^-5 the rate of keratan sulfateβ-galactosidase cleaves lactose into its monosaccharide components, glucose and galactose. It also catalyses the transglycosylation of glucose into allolactose, the inducer of β-galactosidase, in a feedback loop.

Packaging

0.5 unit in glass insert

Physical form

Aseptically filled solution in 20 mM Tris-HCl, pH 7.5

Unit Definition

One unit will release 1.0 µmole of reducing sugar from bovine corneal keratan sulfate per minute at 37 °C, pH 5.8.