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Literature summary extracted from

  • Blixt, O.; van Die, I.; Norberg, T.; van den Eijnden, D.H.
    High-level expression of the Neisseria meningitidis lgtA gene in Escherichia coli and characterization of the encoded N-acetylglucosaminyltransferase as a useful catalyst in the synthesis of GlcNAcb1->3Gal and GalNAcb1->3Gal linkages (1999), Glycobiology, 9, 1061-1071.
    View publication on PubMed

Application

EC Number Application Comment Organism
2.4.1.56 synthesis enzyme promise to be a useful catalyst in the preparation of both GlcNAcbeta1-3Gal and GalNAc1-3Gal linkages Neisseria meningitidis

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.4.1.56 cloning and overexpression of lgtA gene encoding enzyme in Escherichia coli BL21(DE3)/pLys, polypeptide of 333 amino acids Neisseria meningitidis

Inhibitors

EC Number Inhibitors Comment Organism Structure
2.4.1.56 additional information not inhibited by UMP Neisseria meningitidis
2.4.1.56 UDP 1.5 mM: 50% inhibition Neisseria meningitidis

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.4.1.56 0.22
-
 UDP-N-acetyl-D-glucosamine
-
 Neisseria meningitidis
2.4.1.56 2.4
-
 UDP-N-acetyl-D-galactosamine
-
 Neisseria meningitidis
2.4.1.56 4.3
-
 Galbeta1-4Glcbeta-O-4-nitrophenol UDP-N-acetyl-D-glucosamine as donor Neisseria meningitidis
2.4.1.56 4.5
-
 Galbeta1-4Glcbeta-O-4-nitrophenol lipooligosaccharide UDP-N-acetyl-D-galactosamine as donor Neisseria meningitidis
2.4.1.56 6.1
-
 Galbeta1-4GlcNAcbeta-O-4-nitrophenol lipooligosaccharide UDP-N-acetyl-D-glucosamine as donor Neisseria meningitidis
2.4.1.56 7.2
-
 Galbeta1-4GlcNAcbeta-O-4-nitrophenol lipooligosaccharide UDP-N-acetyl-D-galactosamine as donor Neisseria meningitidis
2.4.1.56 27
-
 Galbeta1-3GalNAcbeta-O-4-nitrophenol UDP-N-acetyl-D-glucosamine as donor Neisseria meningitidis
2.4.1.56 28
-
 Galbeta1-4Glc lipooligosaccharide UDP-N-acetyl-D-glucosamine as donor Neisseria meningitidis
2.4.1.56 36
-
 Galbeta1-4Glc lipooligosaccharide UDP-N-acetyl-D-galactosamine as donor Neisseria meningitidis

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
2.4.1.56 Fe2+ activates, less effective than Mn2+ Neisseria meningitidis
2.4.1.56 Mg2+ activates, less effective than Mn2+ Neisseria meningitidis
2.4.1.56 Mn2+ absolute requirement for divalent cations of which Mn2+ is most active, optimal concentration: 15 mM Neisseria meningitidis

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.4.1.56 UDP-N-acetyl-D-glucosamine + lipooligosacccharide Neisseria meningitidis involved in the synthesis of the lacto-N-neo-tetraose structural element of the bacterial lipooligosacccharide, lactose is the natural acceptor disaccharide sequence in lipooligosaccharide UDP + N-acetyl-D-glucosaminyllipooligosaccharide
-
 ?
2.4.1.56 UDP-N-acetyl-D-glucosamine + lipooligosacccharide Neisseria meningitidis H44/76 involved in the synthesis of the lacto-N-neo-tetraose structural element of the bacterial lipooligosacccharide, lactose is the natural acceptor disaccharide sequence in lipooligosaccharide UDP + N-acetyl-D-glucosaminyllipooligosaccharide
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
2.4.1.56 Neisseria meningitidis
-
 immunotype L3
-
2.4.1.56 Neisseria meningitidis H44/76
-
 immunotype L3
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.4.1.56 partial, recombinant enzyme expressed in Escherichia coli Neisseria meningitidis

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
2.4.1.56 0.08 0.09
-
 Neisseria meningitidis

Storage Stability

EC Number Storage Stability Organism
2.4.1.56 -20°C, 50% v/v glycerol, 5mg/ml bovine serum albumin, at least 5 months, stable Neisseria meningitidis

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.4.1.56 additional information no sugar donors: UDPglucose, UDPgalactose, UDPglucosamine, overview over oligosaccharide acceptor specificity Neisseria meningitidis ?
-
 ?
2.4.1.56 additional information no sugar donors: UDPglucose, UDPgalactose, UDPglucosamine, overview over oligosaccharide acceptor specificity Neisseria meningitidis H44/76 ?
-
 ?
2.4.1.56 UDP-N-acetyl-D-galactosamine + lipooligosacccharide 76% of activity with UDP-N-acetyl-D-glucosamine, 10fold lower affinity than for UDP-N-acetyl-D-glucosamine, beta1-3-linkage of N-acetyl-D-galactosamine to accepting Gal residues Neisseria meningitidis UDP + N-acetyl-D-galactosaminyllipooligosaccharide
-
 ?
2.4.1.56 UDP-N-acetyl-D-galactosamine + lipooligosacccharide 76% of activity with UDP-N-acetyl-D-glucosamine, 10fold lower affinity than for UDP-N-acetyl-D-glucosamine, beta1-3-linkage of N-acetyl-D-galactosamine to accepting Gal residues Neisseria meningitidis H44/76 UDP + N-acetyl-D-galactosaminyllipooligosaccharide
-
 ?
2.4.1.56 UDP-N-acetyl-D-glucosamine + Galbeta1-3GalNAcbeta-O-4-nitrophenol
-
 Neisseria meningitidis UDP + GlcNAcbeta-1-3Galbeta1-3GalNAcbeta-O-4-nitrophenol
-
 ?
2.4.1.56 UDP-N-acetyl-D-glucosamine + Galbeta1-3GalNAcbeta-O-p-nitrophenol low affinity Neisseria meningitidis UDP + GlcNAcbeta-1-3Galbeta1-3GalNAcbeta-O-p-nitrophenol
-
 ?
2.4.1.56 UDP-N-acetyl-D-glucosamine + Galbeta1-4Glc lipooligosaccharide low affinity Neisseria meningitidis UDP + GlcNAcbeta1-3Galbeta1-4Glc lipooligosaccharide
-
 ?
2.4.1.56 UDP-N-acetyl-D-glucosamine + Galbeta1-4Glcbeta-O-4-nitrophenol
-
 Neisseria meningitidis UDP + GlcNAcbeta1-3Galbeta1-4Glcbeta-O-4-nitrophenol
-
 ?
2.4.1.56 UDP-N-acetyl-D-glucosamine + Galbeta1-4Glcbeta-O-4-nitrophenol lipooligosaccharide
-
 Neisseria meningitidis UDP + GlcNAcbeta1-3Galbeta1-4Glcbeta-O-4-nitrophenol lipooligosaccharide
-
 ?
2.4.1.56 UDP-N-acetyl-D-glucosamine + Galbeta1-4Glcbeta-O-p-nitrophenol
-
 Neisseria meningitidis UDP + GlcNAcbeta1-3Galbeta1-4Glcbeta-O-p-nitrophenol
-
 ?
2.4.1.56 UDP-N-acetyl-D-glucosamine + Galbeta1-4GlcNAcbeta-O-4-nitrophenol lipooligosaccharide
-
 Neisseria meningitidis UDP + GlcNAcbeta1-3Galbeta1-4GlcNAcbeta-O-4-nitrophenol lipooligosaccharide
-
 ?
2.4.1.56 UDP-N-acetyl-D-glucosamine + Galbeta1-4GlcNAcbeta-O-p-nitrophenol
-
 Neisseria meningitidis UDP + GlcNAcbeta1-3-Galbeta1-4GlcNAcbeta-O-p-nitrophenol
-
 ?
2.4.1.56 UDP-N-acetyl-D-glucosamine + lactose lactose is the highly preferred acceptor substrate Neisseria meningitidis UDP + beta-D-GlcNAc-(1->4)-beta-D-Gal-(1->4)-beta-D-Glc
-
 ?
2.4.1.56 UDP-N-acetyl-D-glucosamine + lactose lactose is the highly preferred acceptor substrate Neisseria meningitidis H44/76 UDP + beta-D-GlcNAc-(1->4)-beta-D-Gal-(1->4)-beta-D-Glc
-
 ?
2.4.1.56 UDP-N-acetyl-D-glucosamine + lipooligosacccharide involved in the synthesis of the lacto-N-neo-tetraose structural element of the bacterial lipooligosacccharide, terminal Gal is an absolute acceptor requirement, broad acceptor specificity toward both alpha- and beta-galactosides structurally related to N- or O-protein or lipid-linked oligosaccharides, beta1-3-linkage of N-acetyl-D-glucosamine to accepting Gal residues in the extending lipooligosacccharide, lactose is the highly preferred acceptor substrate, enzyme also acts efficiently on monomeric and dimeric N-acetyllactosamine Neisseria meningitidis UDP + N-acetyl-D-glucosaminyllipooligosaccharide
-
 ?
2.4.1.56 UDP-N-acetyl-D-glucosamine + lipooligosacccharide involved in the synthesis of the lacto-N-neo-tetraose structural element of the bacterial lipooligosacccharide, lactose is the natural acceptor disaccharide sequence in lipooligosaccharide Neisseria meningitidis UDP + N-acetyl-D-glucosaminyllipooligosaccharide
-
 ?
2.4.1.56 UDP-N-acetyl-D-glucosamine + lipooligosacccharide involved in the synthesis of the lacto-N-neo-tetraose structural element of the bacterial lipooligosacccharide, terminal Gal is an absolute acceptor requirement, broad acceptor specificity toward both alpha- and beta-galactosides structurally related to N- or O-protein or lipid-linked oligosaccharides, beta1-3-linkage of N-acetyl-D-glucosamine to accepting Gal residues in the extending lipooligosacccharide, lactose is the highly preferred acceptor substrate, enzyme also acts efficiently on monomeric and dimeric N-acetyllactosamine Neisseria meningitidis H44/76 UDP + N-acetyl-D-glucosaminyllipooligosaccharide
-
 ?
2.4.1.56 UDP-N-acetyl-D-glucosamine + lipooligosacccharide involved in the synthesis of the lacto-N-neo-tetraose structural element of the bacterial lipooligosacccharide, lactose is the natural acceptor disaccharide sequence in lipooligosaccharide Neisseria meningitidis H44/76 UDP + N-acetyl-D-glucosaminyllipooligosaccharide
-
 ?
2.4.1.56 UDP-N-acetyl-D-glucosamine + N-acetyllactosamine enzyme also acts efficiently on monomeric and dimeric N-acetyllactosamine Neisseria meningitidis UDP + N-acetyl-D-glucosaminyl-N-acetyllactosamine
-
 ?

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.4.1.56 20 30
-
 Neisseria meningitidis
2.4.1.56 30
-
 assay at Neisseria meningitidis

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.4.1.56 7.5 8
-
 Neisseria meningitidis

pH Range

EC Number pH Minimum pH Maximum Comment Organism
2.4.1.56 additional information
-
 broad pH-range within enzyme is active Neisseria meningitidis