BRENDA - Enzyme Database

Enzymatic synthesis of lipopolysaccharide in Escherichia coli. Purification and properties of heptosyltransferase I

Kadrmas, J.L.; Raetz, C.R.; J. Biol. Chem. 273, 2799-2807 (1998)

Data extracted from this reference:

Activating Compound
EC Number
Activating Compound
Commentary
Organism
Structure
2.4.99.B6
Triton X-100
the enzymatic activity is dependent upon the presence of Triton X-100. At concentrations greater than 0.1% Triton X-100, the transferase activity is inhibited
Escherichia coli
Cloned(Commentary)
EC Number
Commentary
Organism
2.4.99.B6
-
Escherichia coli
Inhibitors
EC Number
Inhibitors
Commentary
Organism
Structure
2.4.99.B6
Triton X-100
the enzymatic activity is dependent upon the presence of Triton X-100. At concentrations greater than 0.1% Triton X-100, the transferase activity is inhibited
Escherichia coli
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
2.4.99.B6
0.0045
-
(3-deoxy-alpha-D-manno-oct-2-ulopyranosylonate)-(2->4)-(3-deoxy-alpha-D-manno-oct-2-ulopyranosylonate)-(2->6)-2-deoxy-2-[[(3R)-3-hydroxytetradecanoyl]amino]-3-O-[(3R)-3-hydroxytetradecanoyl]-4-O-phosphono-beta-D-glucopyranosyl-(1->6)-2-deoxy-3-O-[(3R)-3-hydroxytetradecanoyl]-2-[[(3R)-3-hydroxytetradecanoyl]amino]-1-O-phosphono-alpha-D-glucopyranose
pH 7.5, 30°C
Escherichia coli
2.4.99.B6
1.5
-
ADP-alpha-D-mannose
pH 7.5, 30°C
Escherichia coli
Localization
EC Number
Localization
Commentary
Organism
GeneOntology No.
Textmining
2.4.99.B6
membrane
-
Escherichia coli
16020
-
Molecular Weight [Da]
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
2.4.99.B6
36000
-
x * 36000, gel electrophoresis
Escherichia coli
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
2.4.99.B6
Escherichia coli
-
-
-
Purification (Commentary)
EC Number
Commentary
Organism
2.4.99.B6
-
Escherichia coli
Specific Activity [micromol/min/mg]
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
2.4.99.B6
4.9
-
pH 7.5, 30°C
Escherichia coli
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
2.4.99.B6
ADP-alpha-D-mannose + (3-deoxy-alpha-D-manno-oct-2-ulopyranosylonate)-(2->4)-(3-deoxy-alpha-D-manno-oct-2-ulopyranosylonate)-(2->6)-2-deoxy-2-[[(3R)-3-hydroxytetradecanoyl]amino]-3-O-[(3R)-3-hydroxytetradecanoyl]-4-O-phosphono-beta-D-glucopyranosyl-(1->6)-2-deoxy-3-O-[(3R)-3-hydroxytetradecanoyl]-2-[[(3R)-3-hydroxytetradecanoyl]amino]-1-O-phosphono-alpha-D-glucopyranose
i.e. alpha-Kdo-(2->4)-alpha-Kdo-(2->6)-lipid IVA. In cell extracts, ADP-mannose can serve as an alternative donor, but is mannosylated at less than 1% the rate of alpha-Kdo-(2->4)-alpha-Kdo-(2->6)-lipid IVA
727824
Escherichia coli
ADP + ?
-
-
-
?
2.4.99.B6
ADP-L-glycero-beta-D-manno-heptose + (3-deoxy-alpha-D-manno-oct-2-ulopyranosylonate)-(2->4)-(3-deoxy-alpha-D-manno-oct-2-ulopyranosylonate)-(2->6)-2-deoxy-2-[[(3R)-3-hydroxytetradecanoyl]amino]-3-O-[(3R)-3-hydroxytetradecanoyl]-4-O-phosphono-beta-D-glucopyranosyl-(1->6)-2-deoxy-3-O-[(3R)-3-hydroxytetradecanoyl]-2-[[(3R)-3-hydroxytetradecanoyl]amino]-1-O-phosphono-alpha-D-glucopyranose
i.e. alpha-Kdo-(2->4)-alpha-Kdo-(2->6)-lipid IVA. It is possible to assay the enzyme in crude cell extracts using ADP-mannose in place of ADP-L-glycero-beta-D-manno-heptose. No activity without detergent
727824
Escherichia coli
ADP + alpha-L-glycero-D-manno-heptosyl-(1->5)-[(3-deoxy-alpha-D-manno-oct-2-ulopyranosylonate)-(2->4)]-(3-deoxy-alpha-D-manno-oct-2-ulopyranosylonate)-(2->6)-2-deoxy-2-[[(3R)-3-hydroxytetradecanoyl]amino]-3-O-[(3R)-3-hydroxytetradecanoyl]-4-O-phospho-beta-D-glucopyranosyl-(1->6)-2-deoxy-3-O-[(3R)-3-hydroxytetradecanoyl]-2-[[(3R)-3-hydroxytetradecanoyl]amino]-1-O-phosphono-alpha-D-glucopyranose
-
-
-
?
2.4.99.B6
additional information
the purified enzyme displays no activity with ADP-glucose, GDP-mannose, UDP-glucose, or UDP-galactose
727824
Escherichia coli
?
-
-
-
-
Subunits
EC Number
Subunits
Commentary
Organism
2.4.99.B6
?
x * 36000, gel electrophoresis
Escherichia coli
Temperature Optimum [°C]
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
2.4.99.B6
30
-
assay at
Escherichia coli
pH Optimum
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
2.4.99.B6
7.5
-
-
Escherichia coli
pH Range
EC Number
pH Minimum
pH Maximum
Commentary
Organism
2.4.99.B6
7.2
8
pH 7.2: about 70% of maximal activity, pH 8.0: about 55% of maximal activity
Escherichia coli
Activating Compound (protein specific)
EC Number
Activating Compound
Commentary
Organism
Structure
2.4.99.B6
Triton X-100
the enzymatic activity is dependent upon the presence of Triton X-100. At concentrations greater than 0.1% Triton X-100, the transferase activity is inhibited
Escherichia coli
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
2.4.99.B6
-
Escherichia coli
Inhibitors (protein specific)
EC Number
Inhibitors
Commentary
Organism
Structure
2.4.99.B6
Triton X-100
the enzymatic activity is dependent upon the presence of Triton X-100. At concentrations greater than 0.1% Triton X-100, the transferase activity is inhibited
Escherichia coli
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
2.4.99.B6
0.0045
-
(3-deoxy-alpha-D-manno-oct-2-ulopyranosylonate)-(2->4)-(3-deoxy-alpha-D-manno-oct-2-ulopyranosylonate)-(2->6)-2-deoxy-2-[[(3R)-3-hydroxytetradecanoyl]amino]-3-O-[(3R)-3-hydroxytetradecanoyl]-4-O-phosphono-beta-D-glucopyranosyl-(1->6)-2-deoxy-3-O-[(3R)-3-hydroxytetradecanoyl]-2-[[(3R)-3-hydroxytetradecanoyl]amino]-1-O-phosphono-alpha-D-glucopyranose
pH 7.5, 30°C
Escherichia coli
2.4.99.B6
1.5
-
ADP-alpha-D-mannose
pH 7.5, 30°C
Escherichia coli
Localization (protein specific)
EC Number
Localization
Commentary
Organism
GeneOntology No.
Textmining
2.4.99.B6
membrane
-
Escherichia coli
16020
-
Molecular Weight [Da] (protein specific)
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
2.4.99.B6
36000
-
x * 36000, gel electrophoresis
Escherichia coli
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
2.4.99.B6
-
Escherichia coli
Specific Activity [micromol/min/mg] (protein specific)
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
2.4.99.B6
4.9
-
pH 7.5, 30°C
Escherichia coli
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
2.4.99.B6
ADP-alpha-D-mannose + (3-deoxy-alpha-D-manno-oct-2-ulopyranosylonate)-(2->4)-(3-deoxy-alpha-D-manno-oct-2-ulopyranosylonate)-(2->6)-2-deoxy-2-[[(3R)-3-hydroxytetradecanoyl]amino]-3-O-[(3R)-3-hydroxytetradecanoyl]-4-O-phosphono-beta-D-glucopyranosyl-(1->6)-2-deoxy-3-O-[(3R)-3-hydroxytetradecanoyl]-2-[[(3R)-3-hydroxytetradecanoyl]amino]-1-O-phosphono-alpha-D-glucopyranose
i.e. alpha-Kdo-(2->4)-alpha-Kdo-(2->6)-lipid IVA. In cell extracts, ADP-mannose can serve as an alternative donor, but is mannosylated at less than 1% the rate of alpha-Kdo-(2->4)-alpha-Kdo-(2->6)-lipid IVA
727824
Escherichia coli
ADP + ?
-
-
-
?
2.4.99.B6
ADP-L-glycero-beta-D-manno-heptose + (3-deoxy-alpha-D-manno-oct-2-ulopyranosylonate)-(2->4)-(3-deoxy-alpha-D-manno-oct-2-ulopyranosylonate)-(2->6)-2-deoxy-2-[[(3R)-3-hydroxytetradecanoyl]amino]-3-O-[(3R)-3-hydroxytetradecanoyl]-4-O-phosphono-beta-D-glucopyranosyl-(1->6)-2-deoxy-3-O-[(3R)-3-hydroxytetradecanoyl]-2-[[(3R)-3-hydroxytetradecanoyl]amino]-1-O-phosphono-alpha-D-glucopyranose
i.e. alpha-Kdo-(2->4)-alpha-Kdo-(2->6)-lipid IVA. It is possible to assay the enzyme in crude cell extracts using ADP-mannose in place of ADP-L-glycero-beta-D-manno-heptose. No activity without detergent
727824
Escherichia coli
ADP + alpha-L-glycero-D-manno-heptosyl-(1->5)-[(3-deoxy-alpha-D-manno-oct-2-ulopyranosylonate)-(2->4)]-(3-deoxy-alpha-D-manno-oct-2-ulopyranosylonate)-(2->6)-2-deoxy-2-[[(3R)-3-hydroxytetradecanoyl]amino]-3-O-[(3R)-3-hydroxytetradecanoyl]-4-O-phospho-beta-D-glucopyranosyl-(1->6)-2-deoxy-3-O-[(3R)-3-hydroxytetradecanoyl]-2-[[(3R)-3-hydroxytetradecanoyl]amino]-1-O-phosphono-alpha-D-glucopyranose
-
-
-
?
2.4.99.B6
additional information
the purified enzyme displays no activity with ADP-glucose, GDP-mannose, UDP-glucose, or UDP-galactose
727824
Escherichia coli
?
-
-
-
-
Subunits (protein specific)
EC Number
Subunits
Commentary
Organism
2.4.99.B6
?
x * 36000, gel electrophoresis
Escherichia coli
Temperature Optimum [°C] (protein specific)
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
2.4.99.B6
30
-
assay at
Escherichia coli
pH Optimum (protein specific)
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
2.4.99.B6
7.5
-
-
Escherichia coli
pH Range (protein specific)
EC Number
pH Minimum
pH Maximum
Commentary
Organism
2.4.99.B6
7.2
8
pH 7.2: about 70% of maximal activity, pH 8.0: about 55% of maximal activity
Escherichia coli
General Information
EC Number
General Information
Commentary
Organism
2.4.99.B6
malfunction
lipopolysaccharide isolated from mutants defective in rfaC lack heptose and all other sugars distal to heptose
Escherichia coli
General Information (protein specific)
EC Number
General Information
Commentary
Organism
2.4.99.B6
malfunction
lipopolysaccharide isolated from mutants defective in rfaC lack heptose and all other sugars distal to heptose
Escherichia coli