Information on EC 2.4.99.12 - lipid IVA 3-deoxy-D-manno-octulosonic acid transferase

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The expected taxonomic range for this enzyme is: Bacteria, Eukaryota

EC NUMBER
COMMENTARY hide
2.4.99.12
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RECOMMENDED NAME
GeneOntology No.
lipid IVA 3-deoxy-D-manno-octulosonic acid transferase
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REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
lipid IVA + CMP-beta-Kdo = alpha-Kdo-(2->6)-lipid IVA + CMP
show the reaction diagram
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
Kdo transfer to lipid IVA I
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Kdo transfer to lipid IVA II
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lipid A biosynthesis
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Lipopolysaccharide biosynthesis
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Metabolic pathways
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SYSTEMATIC NAME
IUBMB Comments
CMP-3-deoxy-D-manno-oct-2-ulosonate:lipid IVA 3-deoxy-D-manno-oct-2-ulosonate transferase
The bifunctional enzyme from Escherichia coli transfers two 3-deoxy-D-manno-oct-2-ulosonate residues to lipid IVA (cf. EC 2.4.99.13 [(Kdo)-lipid IVA 3-deoxy-D-manno-octulosonic acid transferase]) [1]. The monofunctional enzymes from Aquifex aeolicus and Hemophilus influenzae catalyse the transfer of a single 3-deoxy-D-manno-oct-2-ulosonate residue from CMP-3-deoxy-D-manno-oct-2-ulosonate to lipid IVA [2,3]. The enzymes from Chlamydia transfer three or more 3-deoxy-D-manno-oct-2-ulosonate residues and generate genus-specific epitopes [4].
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
ecotype Columbia
SwissProt
Manually annotated by BRENDA team
strain TW-183
SwissProt
Manually annotated by BRENDA team
strain TW-183
SwissProt
Manually annotated by BRENDA team
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SwissProt
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
malfunction
metabolism
the enzyme is involved in the synthesis of a mitochondrial not yet identified lipid A-like molecule rather than in the synthesis of the cell wall rhamnogalacturonan II
physiological function
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
2-deoxy-2-[[(3R)-3-hydroxypentadecanoyl]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 + CMP-3-deoxy-D-manno-octulosonate
3-deoxy-alpha-D-manno-oct-2-ulopyranosyl-(2->6)-2-deoxy-2-[[(3R)-3-hydroxypentadecanoyl]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 + CMP
show the reaction diagram
lipid IVA + CMP-alpha-Kdo
alpha-Kdo-(2->6)-lipid IVA + CMP
show the reaction diagram
tetraacyl-1,4-bisphosphate lipid A precursor 406 + CMP-alpha-Kdo
CMP + ?
show the reaction diagram
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-
-
?
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
2-deoxy-2-[[(3R)-3-hydroxypentadecanoyl]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 + CMP-3-deoxy-D-manno-octulosonate
3-deoxy-alpha-D-manno-oct-2-ulopyranosyl-(2->6)-2-deoxy-2-[[(3R)-3-hydroxypentadecanoyl]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 + CMP
show the reaction diagram
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
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the enzyme does not require Mg2+
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
polymixin B
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Re endotoxin
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additional information
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no inhibtion by EDTA
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ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
Triton X-100
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stimulates activity
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.052
2-deoxy-2-[[(3R)-3-hydroxypentadecanoyl]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
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pH 8.0, 30°C. 2-deoxy-2-[[(3R)-3-hydroxypentadecanoyl]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 = lipid IV(A)
0.088
CMP-3-deoxy-D-manno-octulosonate
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pH 8.0, 30°C
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6 - 8.5
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pH 6.0: about 45% of maximal activity, pH 8.5: about 45% of maximal activity
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
highest expression
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
PDB
SCOP
CATH
ORGANISM
UNIPROT
Aquifex aeolicus (strain VF5)
Aquifex aeolicus (strain VF5)
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
substrate-free form and in complex with CMP, hanging drop vapor diffusion method, using 100 mM Tris-HCl (pH 8.5), 35-40% (v/v) PEG 400, 200 mM Na-citrate, 50 mM 2-mercaptoethanol, at 19°C
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
30
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unstable
80
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16 h, no effect
90
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incubation for 1 h results in gradual loss of activity
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
HiTrap Heparin-Sepharose column chromatography
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expressed in a rough mutant (Re chemotype) of Escherichia coli (strain F515) that contains an lipopolysaccharide with only two alpha 2-4-linked Kdo residues. Recombinant strains are able to add the immunodominant Kdo residue in alpha 2-8-linkage to the parental lipopolysaccharide. Comparison of nucleotide and the deduced amino acid sequences of gseA of Chlamydia psittaci 6BC and Chlamydia trachomatis L
expressed in Corynebacterium glutamicum
expressed in Escherichia coli CMR300 cells lacking Kdo-transferase
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expression in Escherichia coli. Chlamydial Kdo transferases can replace in Escherichia coli K-12 the host's Kdo transferase and retain the product specificities described in their natural background. WaaA from Chlamydia psittaci transfers predominantly four Kdo residues to lipid A, forming a branched tetrasaccharide with the structure alpha-Kdo-(2,8)-[alpha-Kdo-(2,4)]-alpha-Kdo-(2,4)-alpha-Kdo
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heterologous expression of the Aquifex aeolicus waaA gene in a newly constructed Escherichia coli waaA suppressor strain results in synthesis of lipid IV(A) precursors substituted with one Kdo sugar
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introduction of gseA into an Escherichia coli mutant with a thermolabile kdtA gene product endows cell extracts with the ability to transfer not only the third but also the first two Kdos to lipid A precursors, the Chlamydia trachomatis enzyme is at least trifunctional
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the gene is expressed in the Gram-positive host Corynebacterium glutamicum. Both Escherichia coli strains which express waaA and kdkA from Haemophilus influenzae synthesize an lipopolysaccharide containing a single Kdo residue that was exclusively phosphorylated at position 4
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
E276A
the mutation causes a decrease in the catalytic activity of the enzyme
G30A
the mutant is completely inactive
H272A/N273A
the mutations drastically affect the ability of the enzyme to transfer Kdo
R212A
the mutation clearly affects but does not completely prevent Kdo transfer activity
Show AA Sequence (310 entries)
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