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Literature summary for 1.1.1.22 extracted from

  • Mainprize, I.L.; Bean, J.D.; Bouwman, C.; Kimber, M.S.; Whitfield, C.
    The UDP-glucose dehydrogenase of Escherichia coli K-12 displays substrate inhibition by NAD that is relieved by nucleotide triphosphates (2013), J. Biol. Chem., 288, 23064-23074.
    View publication on PubMedView publication on EuropePMC

Activating Compound

Activating Compound Comment Organism Structure
ADP slight activation Escherichia coli
AMPPNP
-
Escherichia coli
ATP
-
Escherichia coli
ATP highly activating Escherichia coli
dATP highly activating Escherichia coli
diphosphate
-
Escherichia coli
additional information the enzyme shows a mechanism of activation that is NTP-dependent, poor activation by UDP-alpha-D-glucose. The inclusion of MgCl2 in the reaction buffer reverses the NTP-dependent activation of UgdK-12, in a concentration-dependent manner. This is not specific to Mg2+ as a similar effect is found for other divalent cations, including Ca2+, Mn2+, Zn2+, and Ni2+. Monovalent cations do not affect UgdK-12 activity. Chelating the divalent cation with EDTA or EGTA restores the NTP-dependent activation of UgdK-12 Escherichia coli
UDP slight activation Escherichia coli
UTP best activating nucleotide triphosphate Escherichia coli

Cloned(Commentary)

Cloned (Comment) Organism
gene ugd, located near the colanic acid/group 1 CPS locus, recombinant overexpression of His6-tagged enzyme in Escherichia coli strain CWG876 Escherichia coli
gene ugd, located near the colanic acid/group 1 CPS locus, recombinant overexpression of His6-tagged full-length enzyme and truncated enzyme comprising residues 447-704 in Escherichia coli strain CWG876 Escherichia coli

Protein Variants

Protein Variants Comment Organism
K323A site-directed mutagenesis, the mutant shows altered kinetics compared to the wild-type enzyme Escherichia coli
additional information enzyme Ugd from Escherichia coli K-12 can functionally replace enzyme Ugd from Escherichia coli serotype K30 in biosynthesis of K30 capsular polysaccharide Escherichia coli
R324A site-directed mutagenesis, the mutant purifies in much lower amounts relative to wild-type and is prone to degradation and has negligible activity Escherichia coli
Y71F site-directed mutagenesis, the mutant shows unaltered catalytic activity Escherichia coli

Inhibitors

Inhibitors Comment Organism Structure
additional information the enzyme from serotype K30 is not inhibited by NAD+, in contrast to Escherichia coli K-12 Escherichia coli
NAD+ substrate inhibition, reversible by the addition of a nucleotide triphosphate, e.g. ATP, in the absence of kinase. Mutations in a previously identified UDP-glucuronic acid allosteric binding site decrease the binding affinity of the nucleotide triphosphate Escherichia coli
UDP-alpha-D-glucuronate slight product inhibition Escherichia coli

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
additional information Michaelis-Menten kinetics Escherichia coli
0.31
-
NAD+ pH 9.5, 37°C, recombinant wild-type enzyme Escherichia coli
0.35 0.62 NAD+ pH 9.5, 37°C, recombinant wild-type enzyme, from different strains Escherichia coli
0.35 0.62 NAD+ pH 9.5, 37°C, recombinant wild-type enzyme, with 0.025-1.0 mM ATP Escherichia coli
0.43
-
NAD+ pH 9.5, 37°C, recombinant mutant K323A Escherichia coli
0.5
-
NAD+ pH 9.5, 37°C, recombinant mutant Y71F Escherichia coli

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
UDP-alpha-D-glucose + 2 NAD+ + H2O Escherichia coli
-
UDP-alpha-D-glucuronate + 2 NADH + 2 H+
-
?

Organism

Organism UniProt Comment Textmining
Escherichia coli
-
several strains
-
Escherichia coli
-
serotype K30, several strains
-

Posttranslational Modification

Posttranslational Modification Comment Organism
phosphoprotein possible role of Ugd phosphorylation in the production of a constitutively expressed capsular polysaccharide, no role for phosphorylation in modulating activity of Ugd. Escherichia coli might contain two bacterial tyrosine kinases, analysis of kinase deletion mutants of Escherichia coli strains, overview Escherichia coli
phosphoprotein tyrosine phosphorylation in the activation of Ugd of Escherichia coli K12. Possible role of Ugd phosphorylation in the production of a constitutively expressed capsular polysaccharide (CPS), no role for phosphorylation in modulating activity of Ugd. Escherichia coli might contain two bacterial tyrosine kinases, analysis of kinase deletion mutants of Escherichia coli strains, overview Escherichia coli

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
UDP-alpha-D-glucose + 2 NAD+ + H2O
-
Escherichia coli UDP-alpha-D-glucuronate + 2 NADH + 2 H+
-
?

Subunits

Subunits Comment Organism
More structural modelling of the enzyme in complex with NAD and uridine 5'-monophosphate, using structure of Ugd from Klebsiella pneumoniae in complex with UDPGA, PDB ID 3PJG, as template Escherichia coli

Synonyms

Synonyms Comment Organism
UDP-glucose dehydrogenase
-
Escherichia coli
Ugd
-
Escherichia coli

Temperature Optimum [°C]

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

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.7
-
NAD+ pH 9.5, 37°C, recombinant mutant K323A Escherichia coli
4.2
-
NAD+ pH 9.5, 37°C, recombinant wild-type enzyme Escherichia coli
5.4 7.1 NAD+ pH 9.5, 37°C, recombinant wild-type enzyme, from different strains Escherichia coli
7.3
-
NAD+ pH 9.5, 37°C, recombinant mutant Y71F Escherichia coli
56.2
-
NAD+ pH 9.5, 37°C, recombinant wild-type enzyme, with 0.025-1.0 mM ATP Escherichia coli

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
9.5
-
assay at Escherichia coli

Cofactor

Cofactor Comment Organism Structure
NAD+
-
Escherichia coli

General Information

General Information Comment Organism
additional information enzyme Ugd from Escherichia coli K-12 can functionally replace enzyme Ugd from Escherichia coli serotype K30 in biosynthesis of K30 capsular polysaccharide Escherichia coli
physiological function in Escherichia coli K-12, Ugd is important for biosynthesis of the environmentally regulated exopolysaccharide known as colanic acid, whereas in other Escherichia coli isolates, the same enzyme is required for production of the constitutive group 1 capsular polysaccharides, which act as virulence determinants Escherichia coli
physiological function in Escherichia coli serotype K30, the enzyme is required for production of the constitutive group 1 capsular polysaccharides, which act as virulence determinants Escherichia coli