Recombinant Enterococcus faecalis O-glycosidase, E. coli

Synonym: Enterococcus faecalis O-glycosidase

Product Size

2,000 KU; 4,000 KU; 6,000 KU; 8,000 KU

Price

Datasheet

Datasheet

MSDS

MSDS

Properties

Description

Recombinant Enterococcus faecalis O-glycosidase expressed in E. coli catalyzes the removal of Core 1 and Core 3 O-linked disaccharides from glycoproteins.

Unit Definition

One unit will remove 0.68 nmol of O-linked disaccharide from 5 mg of neuraminidase digested, non-denatured fetuin in 1 hour at 37°C in a total reaction volume of 100 µl. One unit of both O-glycosidase and PNGase F will remove equivalent molar amounts of O-linked disaccharides and N-linked oligosaccharides, respectively.

Expression System

E. coli

Source

Enterococcus faecalis

Components

O-Glycosidase (40,000,000 units/ml); 10 x GlycoBuffer (50 mM sodium phosphate, pH 7.5, 25°C); 10 x Glycoprotein denaturing buffer (0.5% SDS, 40 mM DTT); NP-40 (10%)

Reaction Conditions

1X GlycoBuffer (50 mM sodium phosphate, pH 7.5, 25°C), Incubate at 37°C

Heat Inactivation

10 minutes at 65°C

Production Methods

Fermentation

Detection Method

A two-fold gradient dilution of O-glycosidase is added to a reaction mixture of 5 mg of neuraminidase-digested fetuin and 1X GlycoBuffer (50 mM sodium phosphate, pH 7.5, 25°C). The reaction mixture is incubated at 37°C for 1 h. O-linked disaccharide carbohydrates are analyzed by the Morgan and Elson assay.

Concentration

40,000,000 units/ml

Form

Powder

Purity

≥95%, determined by SDS-PAGE.

Buffer

20 mM Tris-HCl, 50 mM NaCl, 1 mM EDTA, pH 7.5 at 25°C

Applications

Recombinant Enterococcus faecalis O-glycosidase expressed in E. coli can be used to remove more complex O-glycan recombinases.

Recombinant Enterococcus faecalis O-glycosidase, E. coli