Endoglycosidases

Endoglycosidases are enzymes that cleave intact glycans from glycoproteins. These enzymes are highly stable in clean preparations with no glycerol, NaCl, or other additives such as EDTA. All enzymes are tested for the absence of proteolytic or unexpected glycosidic activity.

Endoglycosidases are some of the most widely used enzymes for the analysis of glycoproteins. These enzymes release intact glycans from glycoproteins, while exoglycosidases release individual monosaccharides (i.e. sialic acid, galactose, β-N-acetylgucosamine, mannose, etc.).

PNGase F is commonly included in the group of enzymes due to its similar activity of cleaving intact N-linked glycans, though it is actually an amidase. It cleaves between the asparagine amino acid and the first β-N-acetylglucoseamine sugar (GlcNAc) of the chitobiose core of N-linked glycans. The Endo F enzymes and Endo H cleave N-linked glycans between the first and second GlcNAc, leaving a charged residue which aids in increasing solubility of the deglycosylated protein, reducing the potential for precipitation of the protein aggregates. PNGase F cleaves all N-linked glycans, while Endo H and the Endo F enzymes remove specific types of N-linked glycans.

Also included in this classification of enzymes is O-Glycosidase, which removes the Core 1 O-Linked glycan (Gal-β(1-3)GalNAc-α) from serine or threonine amino acids. Often times O-linked glycans contain additional monosaccharides linked to the galactose in both branched and linear linkages, requiring the use of exoglycosidases such as sialidase and galactosidase to remove the additional sugars.

PNGase F (Peptide N Glycosidase F) and recombinant PNGase F

PNGase F is suitable for release of all types (high-mannose, hybrid and complex) N-linked glycans from glycoproteins and glycopeptides. PNGase F will not remove oligosaccharides containing α(1-3) linked core fucose commonly found on plant glycoproteins.

Table below lists and compares the PNGase F enzyme kits offered at Ludger.

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Endoglycosidase F1

E-EF01

£415.00

Endo F1 cleaves high mannose and some hybrid type N-glycans from peptides and proteins.

Endoglycosidase F2

E-EF02

£415.00

Endo F2 cleaves N-linked (asparagine-linked) biantennary oligosaccharides from glycoproteins. It also will cleave high mannose glycans but at a 40x reduced rate. It cleaves between the two N-acetylglucosamine residues in the diacetylchitobiose core of the oligosaccharide, generating a truncated sugar molecule with one N-acetylglucosamine residue remaining on the asparagine. In contrast, PNGase F removes the oligosaccharide intact.

Endoglycosidase F3

E-EF03

£415.00

Selective release of triantennarry and α(1-6) fucosylated biantennary N-glycans from peptides and protein.

Endoglycosidase H

E-EH02

£415.00

Endo H cleaves Asparagine-linked hybrid or high mannose oligosaccharides, but not complex oligosaccharides. It cleaves between the two N-acetylglucosamine residues in the diacetylchitobiose core of the oligosaccharide, generating a truncated sugar molecule with one N-acetylglucosamine residue remaining on the asparagine. In contrast, PNGase F removes the oligosaccharide intact. Detergent and heat denaturation may increase the rate of cleavage for some glycoproteins.

O-glycosidase

E-G001

£415.00

O-glycosidase cleaves only unsubstituted Gal-β(1-3)GalNAc-α disaccharides attached to the serine or threonine residues of glycoproteins or glycopeptides.

PNGase F (Peptide-N-Glycosidase F)

E-PNG01

£415.00

PNGase F (Peptide N Glycosidase F)

E-PNG01-200

£1,185.00

Recombinant PNGase F

E-rPNG01

£415.00

Endo-β-galactosidase

E-XBG01

£415.00

Endo-β-Galactosidase cleaves internal β(1-4) galactose linkages in unbranched, repeating poly-N-acetyllactosamine structures. Sulfated structures such as keratan sulfate are also cleaved. Branching and/or fucosylation of the substrate may decrease or eliminate cleavage.

Enzymatic CarboRelease kit

KE-DG01

£516.00

This kit includes the enzymes and buffers required to deglycosylate glycoproteins, removing all N-linked oligosaccharides and many O-linked sugars.

Enzymatic DeGlycoMx Kit

KE-DGMX

£296.00

This kit developed for protein deglycosylation includes our DeGlycoMx, a premixed cocktail of the enzymes required to remove all N-linked oligosaccharides and most O-linked sugars from 0.5 mg of glycoprotein, via 10 reactions of up to 50 micrograms of protein per reaction.

Ceramide Glycanase Kit

LZ-CER-HM-KIT

£344.00

Ceramide glycanase can be used to deglycosylate a variety of glycosphingolipids by cleaving the β-glycosyl linkage. Ceramide glycanase allows the identification of glycosylation patterns of glycosphingolipids through cleaving the glycan moiety and making it accessible to LudgerTag labelling technology.

LudgerZyme PNGase L Kit

LZ-PNGaseL-50-KIT

£391.00

Recombinant PNGase F

LZ-rPNGaseF-kit

£672.00

LudgerZyme recombinant PNGase F kit (New England BioLabs).

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