Enzymes

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  • N-acetyl-beta-D-glucosaminide beta-(1,3)-galactosyltransferase     Size: 4 mU

    Introduction: B3GalT is a recombinant N-acetyl-beta-D-glucosaminide beta-(1,3)-galactosyltransferase. The enzyme transfers galactose from UDP-galactose to the N-acetylglucosamine or N-acetylgalactosamine termini of acceptor molecules such as GalNAc, GlcNAc, Lacto-N-triose,  Globotetrose, Isoglobotetrose and GalNAcβ1,3Galβ1,4Glc. Besides, B3GalT has been used to produce the Human Milk Oligosaccharide (HMO) lacto‐N‐tetraose (LNT) and  N-Acetyllactosamine (LacNAc) Type 1 O-glycans.

    B3Gal Transferase can be used for synthesis of  the precursor of H type 1 antigen of ABO histo-blood group. The H type 1 antigen in itself is the precursor of  the following type 1 antigens:  antigens A, antigen B, Lewis a (Le a ), Lewis b (Le b ) and  Sialyl Lewis a (sLe a).

    Source: E. coli cells with a cloned bacterial B3GalT gene.

    Storage Buffer: The enzyme is provided in solution containing Tris buffer, NaCl and 10% glycerol

    Activity: The activity per µL is 40 µUnits/µL or 248 pmol/min/μL

    Milliunit Definition: One milliunit will transfer 1.0 nanomole of galactose from UDP-galactose to 4-Nitrophenyl N-acetyl-β-D-glucosaminide per min at pH 7.5 at 37 °C.

    Storage: Store at -20 °C >1 year. Avoid repeated freeze-thaw cycles, which decrease protein stability.

    764.00
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  • Beta-(1,3)-N-Acetylgalactosaminyltransferase     Size: 10 mU

    Introduction:

    B3GlcNAcT is a β-1,3-N-acetylgalactosaminyltransferase that transfers glucosamine from UDP-glucosamine to the Galactosyl termini of acceptor molecules such as Lactose (Galβ1→4Glc), Lactosamine (Galβ1→4GlcNAc), O-glycans (Galβ1→3GalNAc),  Lacto-N-biose (Galβ1→3GlcNAc), Galβ1→6GlcNAc,  Galβ1→4Xylβ-o-Bn or Biantennary, Triantennary and Tetraantennary N-Glycans.

    It can function also as a B3GlcNAcT (Beta-(1,3)-N-Acetylglucosamilyltransferase)

    Activity: 

    Overnight incubation with 4 µL of this enzyme will produce 1 mg of product. If higher amount of product is needed the enzyme will continuously add GlcNAc for at least one week.
    Reaction conditions are: Donor: UDP-GLcNAc, substrate: pNP-Lactose, product: pNP-Lactose-GlcNAc, reaction temperature: 23 ºC , incubation time: 18 hours.

    The specific activity is 975.7 pmol/min/μg

    Source: E. coli cells with a cloned bacterial B3GlcNAc Transferase gene.

    Storage Buffer: The enzyme is provided as lyophilized powder solution containing Tris buffer and NaCl

    Milliunit Definition: One milliunit will transfer 1.0 nanomole of GlcNAc (N-Acetyl-D-glucosamine) from UDP-GlcNAc (Uridine diphosphate N-acetylglucosamine) to Galβ1→4Glcβ-O-pNP (4-Nitrophenyl β -D-lactopyranoside) per min at pH 7.5 at 25 °C.

    Storage: Store at -20 °C >1 year. Avoid repeated freeze-thaw cycles, which decrease protein stability.

    181.00
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  • Beta-(1,3)-N-Acetylglucosaminyltransferase     Size: 100 mU

    Introduction: B3GlcNAcT is a β-1,3-N-acetylglucosaminyltransferase that transfers glucosamine from UDP-glucosamine to the Galactosyl termini of acceptor molecules such as Lactose (Galβ1→4Glc), Lactosamine (Galβ1→4GlcNAc), O-glycans (Galβ1→3GalNAc),  Lacto-N-biose (Galβ1→3GlcNAc), Galβ1→6GlcNAc,  Galβ1→4Xylβ-o-Bn or Biantennary, Triantennary and Tetraantennary N-Glycans. It can function also as a Beta-(1,3)-N-Acetylgalactosaminyltransferase

    Activity:  Overnight incubation with 4 µL of this enzyme will produce 1 mg of product. If higher amount of product is needed the enzyme will continuously add GlcNAc for at least one week. Reaction conditions are: Donor: UDP-GLcNAc, substrate: pNP-Lactose, product: pNP-Lactose-GlcNAc, reaction temperature: 23 ºC , incubation time: 18 hours. The specific activity is 975.7 pmol/min/μg

    Source: E. coli cells with a cloned bacterial B3GlcNAc Transferase gene.

    Storage Buffer: The enzyme is provided as lyophilized powder solution containing Tris buffer and NaCl

    Milliunit Definition: One milliUnit (mU) is defined as the amount of enzyme that catalyzes the formation of 1 nanomol of 4-Nitrophenyl β -D-lactopyranoside-GlcNAc from UDP-GlcNAc (Uridine diphosphate N-acetylglucosamine) and Galβ1→4Glcβ-O-pNP (4-Nitrophenyl β -D-lactopyranoside) per min at 25 °C and at pH 7.5.

    Storage: Store at -20 °C >1 year. Avoid repeated freeze-thaw cycles, which decrease protein stability.

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  • Introduction: CarboClip is a recombinant peptide-N-Glycosidase F (PNGase F), from Elizabethkingia sp, overexpressed in E. coli that contains a hexahistidine tag in its C-term and cleaves between the GlcNAc and asparagine residues of high mannose, hybrid, and complex oligosaccharides from N-linked glycoproteins. CarboClip is provided glycerol-free as a lyophilized powder containing Tris-HCl, NaCl2, and Na2EDTA. Activity: After reconstitution in 150 μL of UltraPure H2O, 1 μl of CarboClip can deglycosylate > 95% of ribonuclease B (1 mg) in less than 20 minutes. Activity is calculated on a 12 % SDS-PAGE and percent of deglycosylation using densitometry. Storage: Lyophilized powder: -80 °C and -20 °C >1 year. At room temperature 10 days. Reconstituted protein should be stored at 4 °C and is stable for 2-3 months. Glycosylation of Membrane Proteins: Removal of N-Linked Glycans from membrane protein samples using this enzyme has been described by Contessotto et al. Matrix Biol. 2019 (See applications section below)

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