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Literature summary for 2.4.1.305 extracted from

  • Brockhausen, I.; Hu, B.; Liu, B.; Lau, K.; Szarek, W.A.; Wang, L.; Feng, L.
    Characterization of two beta-1,3-glucosyltransferases from Escherichia coli serotypes O56 and O152 (2008), J. Bacteriol., 190, 4922-4932.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
-
Escherichia coli

Inhibitors

Inhibitors Comment Organism Structure
CHAPS 0.5%, 41% inhibition; 0.5%, 75% inhibition Escherichia coli
Co2+ the addition of 5 mM Co2+ reduces the activation by 5 mM MnCl2 of the enzyme by 45%; the addition of 5 mM Co2+ reduces the activation by 5 mM MnCl2 of the enzyme by 76% Escherichia coli
EDTA
-
Escherichia coli
octyl-beta-D-glucoside 0.5%, 15% inhibition; 0.5%, 78% inhibition Escherichia coli
Triton X-100
-
Escherichia coli

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
N-acetyl-alpha-D-glucosaminyl-diphospho-11-phenoxyundecanol it is not possible to determine the accurate Km and Vmax values for the standard acceptor substrate D-GlcNAc-alpha-diphospho-(CH2)11-O-Ph due to its hydrophobic character Escherichia coli
additional information
-
N-acetyl-alpha-D-glucosaminyl-diphospho-ditrans,octacis-undecaprenol it is not possible to determine the accurate Km and Vmax values for the standard acceptor substrate N-acetyl-D-glucosaminyl-alpha-diphospho-11-phenoxyundecaprenol due to its hydrophobic character Escherichia coli
0.14
-
UDP-alpha-D-glucose pH 7.0, 37°C, assayed with bacterial membranes Escherichia coli
0.5
-
UDP-alpha-D-glucose pH 7.0, 37°C, assayed with bacterial membranes Escherichia coli

Localization

Localization Comment Organism GeneOntology No. Textmining
membrane
-
Escherichia coli 16020
-

Metals/Ions

Metals/Ions Comment Organism Structure
Co2+ cobalt(II)-acetate supports glucosyltransferase activity but much less than with MnCl2 Escherichia coli
Mg2+ both MnCl2 and MgCl2 are efficient activators at 5 mM concentration Escherichia coli
Mn2+ both MnCl2 and MgCl2 are efficient activators at 5 mM concentration. The enzyme is fully active at MnCl2 concentrations between 0.5 and 25 mM Escherichia coli
additional information no activation by Ni2+, Cu2+, Pb2+, Zn2+, and Ca2+ Escherichia coli
additional information no activation by Ni2+, Cu2+, Pb2+, Zn2+, and Ca2+ significantly supported the glucosyltransferase activities Escherichia coli

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
55000
-
SDS-PAGE Escherichia coli
55400
-
SDS-PAGE Escherichia coli

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
UDP-alpha-D-glucose + N-acetyl-alpha-D-glucosaminyl-diphospho-ditrans,octacis-undecaprenol Escherichia coli the enzyme catalyzes the formation of the Glc-(1->3)-GlcNAc linkage in the repeating units of the O-antigen oligosaccharide of Escherichia coli serotype O152 UDP + beta-D-Glc-(1->3)-alpha-D-GlcNAc-diphospho-ditrans,octacis-undecaprenol
-
?
UDP-alpha-D-glucose + N-acetyl-alpha-D-glucosaminyl-diphospho-ditrans,octacis-undecaprenol Escherichia coli the enzyme catalyzes the formation of the Glc-1-3-GlcNAc linkage in the repeating units of the O-antigen oligosaccharide Of Escherichia coli serotype O56 UDP + beta-D-Glc-(1->3)-alpha-D-GlcNAc-diphospho-ditrans,octacis-undecaprenol
-
?
UDP-alpha-D-glucose + N-acetyl-alpha-D-glucosaminyl-diphospho-ditrans,octacis-undecaprenol Escherichia coli serotype O56 the enzyme catalyzes the formation of the Glc-1-3-GlcNAc linkage in the repeating units of the O-antigen oligosaccharide Of Escherichia coli serotype O56 UDP + beta-D-Glc-(1->3)-alpha-D-GlcNAc-diphospho-ditrans,octacis-undecaprenol
-
?
UDP-alpha-D-glucose + N-acetyl-alpha-D-glucosaminyl-diphospho-ditrans,octacis-undecaprenol Escherichia coli serotype O152 the enzyme catalyzes the formation of the Glc-(1->3)-GlcNAc linkage in the repeating units of the O-antigen oligosaccharide of Escherichia coli serotype O152 UDP + beta-D-Glc-(1->3)-alpha-D-GlcNAc-diphospho-ditrans,octacis-undecaprenol
-
?

Organism

Organism UniProt Comment Textmining
Escherichia coli B5L3F2
-
-
Escherichia coli Q077R2
-
-
Escherichia coli serotype O152 B5L3F2
-
-
Escherichia coli serotype O56 Q077R2
-
-

Purification (Commentary)

Purification (Comment) Organism
purified protein has no enzyme activity. The native enzyme is associated with membranes which may be essential for their function Escherichia coli

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.0022
-
pH 7.0, 37°C, assayed with bacterial membranes Escherichia coli
0.025
-
pH 7.0, 37°C, assayed with bacterial membranes Escherichia coli

Storage Stability

Storage Stability Organism
-20°C, stable for several weeks Escherichia coli
4°C, stable for several days Escherichia coli

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
additional information no activity with N-butyryl-glucosamine-beta-1-thio-2-naphthyl as acceptor substrate Escherichia coli ?
-
?
additional information no activity with N-butyryl-glucosamine-beta-1-thio-2-naphthyl as acceptor substrate Escherichia coli serotype O56 ?
-
?
additional information no activity with N-butyryl-glucosamine-beta-1-thio-2-naphthyl as acceptor substrate Escherichia coli serotype O152 ?
-
?
UDP-alpha-D-glucose + alpha-D-GlcNAc-diphospho-(CH2)9-CH3 150% of the activity compared to N-acetyl-alpha-D-glucosaminyl-diphospho-11-phenoxyundecaprenol as acceptor substrate Escherichia coli UDP + beta-D-Glc-(1->3)-alpha-D-GlcNAc-diphospho-(CH2)9-CH3
-
?
UDP-alpha-D-glucose + alpha-D-GlcNAc-diphospho-(CH2)9-CH3 150% of the activity compared to N-acetyl-alpha-D-glucosaminyl-diphospho-11-phenoxyundecaprenol as acceptor substrate Escherichia coli serotype O152 UDP + beta-D-Glc-(1->3)-alpha-D-GlcNAc-diphospho-(CH2)9-CH3
-
?
UDP-alpha-D-glucose + alpha-D-GlcNAc-diphospho-16-phenoxyhexadecanol
-
Escherichia coli UDP + beta-D-Glc-(1->3)-alpha-D-GlcNAc-diphospho-16-phenoxyhexadecanol
-
?
UDP-alpha-D-glucose + alpha-D-GlcNAc-diphospho-16-phenoxyhexadecanol
-
Escherichia coli serotype O152 UDP + beta-D-Glc-(1->3)-alpha-D-GlcNAc-diphospho-16-phenoxyhexadecanol
-
?
UDP-alpha-D-glucose + alpha-D-GlcNAc-diphospho-6-phenoxyhexanol
-
Escherichia coli UDP + beta-D-Glc-(1->3)-alpha-D-GlcNAc-diphospho-6-phenoxyhexanol
-
?
UDP-alpha-D-glucose + alpha-D-GlcNAc-diphospho-6-phenoxyhexanol
-
Escherichia coli serotype O152 UDP + beta-D-Glc-(1->3)-alpha-D-GlcNAc-diphospho-6-phenoxyhexanol
-
?
UDP-alpha-D-glucose + N-acetyl-alpha-D-glucosaminyl-diphospho-(CH2)9-CH3 32% of the activity compared to N-acetyl-alpha-D-glucosaminyl-diphospho-11-phenoxyundecaprenol as acceptor substrate Escherichia coli UDP + beta-D-Glc-(1->3)-alpha-D-GlcNAc-diphospho-(CH2)9-CH3
-
?
UDP-alpha-D-glucose + N-acetyl-alpha-D-glucosaminyl-diphospho-(CH2)9-CH3 32% of the activity compared to N-acetyl-alpha-D-glucosaminyl-diphospho-11-phenoxyundecaprenol as acceptor substrate Escherichia coli serotype O56 UDP + beta-D-Glc-(1->3)-alpha-D-GlcNAc-diphospho-(CH2)9-CH3
-
?
UDP-alpha-D-glucose + N-acetyl-alpha-D-glucosaminyl-diphospho-11-phenoxyundecanol the enzyme requires the diphosphate-sugar linkage in the acceptor substrate for full activity. Substrates lacking diphosphate linked to GlcNAc are inactive. An acceptor analog containing a single phosphate group (D-GlcNAc-alpha-diphospho-11-phenoxyundecanol) shows 11% activity. The phenyl group is not required for activity. The length of the aliphatic chain of acceptor substrates also contributes to the activity. The synthetic acceptor N-acetyl-alpha-D-glucosaminyl-diphospho-11-phenoxyundecaprenol is an analog of the natural undecaprenol-pyrophosphate-sugar acceptor substrate. No activity with CMP-sialic acid, UDP-GlcNAc, UDP-GalNAc, or GDP-Man as donor substrate. UDP-Gal shows 0.2% of the activity compared to UDP-D-glucose Escherichia coli UDP + beta-D-Glc-(1->3)-alpha-D-GlcNAc-diphospho-11-phenoxyundecanol
-
?
UDP-alpha-D-glucose + N-acetyl-alpha-D-glucosaminyl-diphospho-11-phenoxyundecanol the enzyme requires the diphosphate-sugar linkage in the acceptor substrate for full activity. Substrates lacking diphosphate linked to GlcNAc are inactive. An acceptor analog containing a single phosphate group (D-GlcNAc-alpha-P-(CH2)11-O-Ph) shows 1% activity. The phenyl group is not required for activity. The length of the aliphatic chain of acceptor substrates also contributes to the activity. The synthetic acceptor N-acetyl-alpha-D-glucosaminyl-diphospho-11-phenoxyundecaprenol is an analog of the natural undecaprenol-diphosphate-sugar acceptor substrate. No activity with CMP-sialic acid, UDP-GlcNAc, UDP-GalNAc, or GDP-Man as donor substrate. UDP-Gal shows 0.6% of the activity compared to UDP-D-glucose Escherichia coli UDP + beta-D-Glc-(1->3)-alpha-D-GlcNAc-diphospho-11-phenoxyundecanol
-
?
UDP-alpha-D-glucose + N-acetyl-alpha-D-glucosaminyl-diphospho-11-phenoxyundecanol the enzyme requires the diphosphate-sugar linkage in the acceptor substrate for full activity. Substrates lacking diphosphate linked to GlcNAc are inactive. An acceptor analog containing a single phosphate group (D-GlcNAc-alpha-P-(CH2)11-O-Ph) shows 1% activity. The phenyl group is not required for activity. The length of the aliphatic chain of acceptor substrates also contributes to the activity. The synthetic acceptor N-acetyl-alpha-D-glucosaminyl-diphospho-11-phenoxyundecaprenol is an analog of the natural undecaprenol-diphosphate-sugar acceptor substrate. No activity with CMP-sialic acid, UDP-GlcNAc, UDP-GalNAc, or GDP-Man as donor substrate. UDP-Gal shows 0.6% of the activity compared to UDP-D-glucose Escherichia coli serotype O56 UDP + beta-D-Glc-(1->3)-alpha-D-GlcNAc-diphospho-11-phenoxyundecanol
-
?
UDP-alpha-D-glucose + N-acetyl-alpha-D-glucosaminyl-diphospho-16-phenoxyhexadecanol
-
Escherichia coli UDP + beta-D-Glc-(1->3)-alpha-D-GlcNAc-diphospho-16-phenoxyhexadecanol
-
?
UDP-alpha-D-glucose + N-acetyl-alpha-D-glucosaminyl-diphospho-16-phenoxyhexadecanol
-
Escherichia coli serotype O56 UDP + beta-D-Glc-(1->3)-alpha-D-GlcNAc-diphospho-16-phenoxyhexadecanol
-
?
UDP-alpha-D-glucose + N-acetyl-alpha-D-glucosaminyl-diphospho-6-phenoxyhexanol
-
Escherichia coli UDP + beta-D-Glc-(1->3)-alpha-D-GlcNAc-diphospho-6-phenoxyhexanol
-
?
UDP-alpha-D-glucose + N-acetyl-alpha-D-glucosaminyl-diphospho-ditrans,octacis-undecaprenol the enzyme catalyzes the formation of the Glc-(1->3)-GlcNAc linkage in the repeating units of the O-antigen oligosaccharide of Escherichia coli serotype O152 Escherichia coli UDP + beta-D-Glc-(1->3)-alpha-D-GlcNAc-diphospho-ditrans,octacis-undecaprenol
-
?
UDP-alpha-D-glucose + N-acetyl-alpha-D-glucosaminyl-diphospho-ditrans,octacis-undecaprenol the enzyme catalyzes the formation of the Glc-1-3-GlcNAc linkage in the repeating units of the O-antigen oligosaccharide Of Escherichia coli serotype O56 Escherichia coli UDP + beta-D-Glc-(1->3)-alpha-D-GlcNAc-diphospho-ditrans,octacis-undecaprenol
-
?
UDP-alpha-D-glucose + N-acetyl-alpha-D-glucosaminyl-diphospho-ditrans,octacis-undecaprenol the enzyme catalyzes the formation of the Glc-1-3-GlcNAc linkage in the repeating units of the O-antigen oligosaccharide Of Escherichia coli serotype O56 Escherichia coli serotype O56 UDP + beta-D-Glc-(1->3)-alpha-D-GlcNAc-diphospho-ditrans,octacis-undecaprenol
-
?
UDP-alpha-D-glucose + N-acetyl-alpha-D-glucosaminyl-diphospho-ditrans,octacis-undecaprenol the enzyme catalyzes the formation of the Glc-(1->3)-GlcNAc linkage in the repeating units of the O-antigen oligosaccharide of Escherichia coli serotype O152 Escherichia coli serotype O152 UDP + beta-D-Glc-(1->3)-alpha-D-GlcNAc-diphospho-ditrans,octacis-undecaprenol
-
?

Subunits

Subunits Comment Organism
? x * 55000, SDS-PAGE Escherichia coli
? x * 55400, SDS-PAGE Escherichia coli

Synonyms

Synonyms Comment Organism
UDP-Glc:GlcNAc-pyrophosphate-lipid beta-1,3-glucosyltransferase
-
Escherichia coli
WfaP
-
Escherichia coli
WfgD
-
Escherichia coli

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
assay at Escherichia coli

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
6
-
-
Escherichia coli
6.5
-
-
Escherichia coli
7
-
assay at Escherichia coli

pH Range

pH Minimum pH Maximum Comment Organism
5 9 pH 5.0: about 60% of maximal activity, pH 9.0: about 40% of maximal activity Escherichia coli
5 9 pH 5.0: about 75% of maximal activity, pH 9.0: about 75% of maximal activity Escherichia coli