Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary for 2.7.7.12 extracted from

  • Geeganage, S.; Ling, V.W.; Frey, P.A.
    Roles of two conserved amino acid residues in the active site of galactose-1-phosphate uridylyltransferase: an essential serine and a nonessential cysteine (2000), Biochemistry, 39, 5397-5404.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
overexpression of wild-type and mutants in strain BL21(DE3)pLysS Escherichia coli

Protein Variants

Protein Variants Comment Organism
C160A site-directed mutagenesis, 18fold reduced activity Escherichia coli
C160S site-directed mutagenesis, 12fold reduced activity Escherichia coli
H166G active site structure, complex of enzyme with UDP-D-galactose Escherichia coli
S161A site-directed mutagenesis, 7000fold reduced activity compared to wild-type Escherichia coli

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
additional information kinetics at 4°C, wild-type and mutants Escherichia coli
0.062
-
UDP-galactose recombinant mutant S161A, pH 8.5, 27°C Escherichia coli
0.072
-
UDP-glucose recombinant mutant C160A, pH 8.5, 27°C Escherichia coli
0.078
-
UDP-galactose recombinant mutant C160A, pH 8.5, 27°C Escherichia coli
0.082
-
UDP-glucose recombinant mutant C160A, pH 8.5, 27°C Escherichia coli
0.12
-
UDP-galactose recombinant wild-type, pH 8.5, 27°C Escherichia coli
0.125
-
alpha-D-galactose 1-phosphate recombinant mutant C160A, pH 8.5, 27°C Escherichia coli
0.14
-
alpha-D-glucose 1-phosphate recombinant mutant C160A, pH 8.5, 27°C Escherichia coli
0.16
-
alpha-D-glucose 1-phosphate recombinant wild-type, pH 8.5, 27°C Escherichia coli
0.2
-
alpha-D-glucose 1-phosphate recombinant mutant S161A, pH 8.5, 27°C Escherichia coli
0.2
-
UDP-glucose recombinant wild-type, pH 8.5, 27°C Escherichia coli
0.223
-
alpha-D-galactose 1-phosphate recombinant mutant S161A, pH 8.5, 27°C Escherichia coli
0.3
-
alpha-D-galactose 1-phosphate recombinant wild-type, pH 8.5, 27°C Escherichia coli

Metals/Ions

Metals/Ions Comment Organism Structure
Fe2+ 0.7 mol per mol of subunit Escherichia coli
Fe2+ mutant C160A: 0.5 mol per mol of subunit, mutant S161A: 0.4 mol per mol of subunit Escherichia coli
Zn2+ 1.2 mol tightly bound per mol of subunit Escherichia coli
Zn2+ mutant C160A: 1.3 mol per mol of subunit, mutant S161A: 1.2 mol per mol of subunit like the wild-type Escherichia coli

Organism

Organism UniProt Comment Textmining
Escherichia coli P09148 galT gene
-

Purification (Commentary)

Purification (Comment) Organism
recombinant wild-type and mutants from strain BL21(DE3)pLysS to near homogeneity Escherichia coli

Reaction

Reaction Comment Organism Reaction ID
UDP-alpha-D-glucose + alpha-D-galactose 1-phosphate = alpha-D-glucose 1-phosphate + UDP-alpha-D-galactose double-displacement mechanism Escherichia coli
UDP-alpha-D-glucose + alpha-D-galactose 1-phosphate = alpha-D-glucose 1-phosphate + UDP-alpha-D-galactose active site structure, enzyme complexed with UDP-galactose Escherichia coli

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.00093
-
recombinant mutant S161A, cell extract Escherichia coli
0.023
-
purified recombinant mutant S161A Escherichia coli
9.9
-
recombinant mutant C160A, cell extract Escherichia coli
14.9
-
recombinant mutant C160S, cell extract Escherichia coli
16.3
-
recombinant wild-type, cell extract Escherichia coli
79.5
-
purified recombinant mutant C160A Escherichia coli
180
-
purified recombinant wild-type enzyme Escherichia coli

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
UDP-glucose + alpha-D-galactose 1-phosphate
-
Escherichia coli alpha-D-glucose 1-phosphate + UDP-galactose
-
r

Synonyms

Synonyms Comment Organism
GALT
-
Escherichia coli

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
283
-
UDP-galactose recombinant wild-type, reverse reaction, pH 8.5, 27°C Escherichia coli
780
-
alpha-D-galactose 1-phosphate recombinant wild-type, forward reaction, pH 8.5, 27°C Escherichia coli