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Literature summary for 1.3.1.98 extracted from

  • Nishida, S.; Kurokawa, K.; Matsuo, M.; Sakamoto, K.; Ueno, K.; Kita, K.; Sekimizu, K.
    Identification and characterization of amino acid residues essential for the active site of UDP-N-acetylenolpyruvylglucosamine reductase (MurB) from Staphylococcus aureus (2006), J. Biol. Chem., 281, 1714-1724.
    View publication on PubMed
Search for more literature for 1.3.1.98

Cloned(Commentary)

Cloned (Comment) Organism
gene murB, overexpression of wild-type and mutant enzymes in Escherichia coli strain BL21(DE3), functional complementation of murB-deficient, temperature-sensitive Staphylcoccus aureus mutant strain TS2901 by the wild-type enzyme, and enzyme mutants G67A, S70A, G69A, and R230A Staphylococcus aureus

Protein Variants

Protein Variants Comment Organism
E296A site-directed mutagenesis, highly reduced activity compared to the wild-type enzyme, no complementation of murB-deficient, temperature-sensitive Staphylcoccus aureus mutant strain TS2901 Staphylococcus aureus
G67A site-directed mutagenesis, functional complementation of murB-deficient, temperature-sensitive Staphylcoccus aureus mutant strain TS2901 Staphylococcus aureus
G69A site-directed mutagenesis, functional complementation of murB-deficient, temperature-sensitive Staphylcoccus aureus mutant strain TS2901 Staphylococcus aureus
H259A site-directed mutagenesis, highly reduced activity compared to the wild-type enzyme, no complementation of murB-deficient, temperature-sensitive Staphylcoccus aureus mutant strain TS2901 Staphylococcus aureus
N71A site-directed mutagenesis, highly reduced activity compared to the wild-type enzyme, no complementation of murB-deficient, temperature-sensitive Staphylcoccus aureus mutant strain TS2901 Staphylococcus aureus
R176A site-directed mutagenesis, highly reduced activity compared to the wild-type enzyme, no complementation of murB-deficient, temperature-sensitive Staphylcoccus aureus mutant strain TS2901 Staphylococcus aureus
R213A site-directed mutagenesis, nearly inactive mutant, no complementation of murB-deficient, temperature-sensitive Staphylcoccus aureus mutant strain TS2901 Staphylococcus aureus
R230A site-directed mutagenesis, functional complementation of murB-deficient, temperature-sensitive Staphylcoccus aureus mutant strain TS2901 Staphylococcus aureus
S226A site-directed mutagenesis, highly reduced activity compared to the wild-type enzyme, no complementation of murB-deficient, temperature-sensitive Staphylcoccus aureus mutant strain TS2901 Staphylococcus aureus
S70A site-directed mutagenesis, functional complementation of murB-deficient, temperature-sensitive Staphylcoccus aureus mutant strain TS2901 Staphylococcus aureus
Y175F site-directed mutagenesis, highly reduced activity compared to the wild-type enzyme, poor functional complementation of murB-deficient, temperature-sensitive Staphylcoccus aureus mutant strain TS2901 Staphylococcus aureus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
 additional information kinetics of recombinant wild-type and mutant enzymes Staphylococcus aureus
0.0033
-
 NADPH pH 8.0, 22°C, recombinant mutant S226A Staphylococcus aureus
0.008
-
 NADPH pH 8.0, 22°C, recombinant mutant N71A Staphylococcus aureus
0.0095
-
 NADPH pH 8.0, 22°C, recombinant mutant Y175F Staphylococcus aureus
0.014
-
 NADPH pH 8.0, 22°C, recombinant mutant E296A Staphylococcus aureus
0.023
-
 NADPH pH 8.0, 22°C, recombinant mutant R176A Staphylococcus aureus
0.024
-
 NADPH pH 8.0, 22°C, recombinant mutant H259A Staphylococcus aureus
0.036
-
 NADPH pH 8.0, 22°C, recombinant mutant R213A Staphylococcus aureus
0.038
-
 NADPH pH 8.0, 22°C, recombinant wild-type enzyme Staphylococcus aureus

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
UDP-N-acetylenolpyruvylglucosamine + NADPH Staphylococcus aureus
-
 UDP-N-acetylmuramic acid + NADP+
-
 ?

Organism

Organism UniProt Comment Textmining
Staphylococcus aureus P61431 gene murB
-

Purification (Commentary)

Purification (Comment) Organism
recombinant wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) to homogeneity, native wild-type enzyme from Staphylococcus aureus 10fold by ammonium sulfate precipitation and two steps of anion exchange chromatography Staphylococcus aureus

Reaction

Reaction Comment Organism Reaction ID
UDP-N-acetyl-alpha-D-muramate + NADP+ = UDP-N-acetyl-3-O-(1-carboxyvinyl)-alpha-D-glucosamine + NADPH + H+ reaction mechanism, detailed scheme of the second half Staphylococcus aureus
a

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
additional information
-
-
 Staphylococcus aureus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
UDP-N-acetylenolpyruvylglucosamine + NADPH
-
 Staphylococcus aureus UDP-N-acetylmuramic acid + NADP+
-
 ?
UDP-N-acetylenolpyruvylglucosamine + NADPH residues Arg213, Arg176, His259, Asn71, Tyr175, Ser226, and Glu296 are involved in catalysis Staphylococcus aureus UDP-N-acetylmuramic acid + NADP+
-
 ?

Synonyms

Synonyms Comment Organism
MurB
-
 Staphylococcus aureus
UDP-N-acetylenolpyruvylglucosamine reductase
-
 Staphylococcus aureus

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
22
-
 assay at room temperature Staphylococcus aureus

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8
-
 assay at Staphylococcus aureus

Cofactor

Cofactor Comment Organism Structure
FAD bound Staphylococcus aureus
NADPH dependent on Staphylococcus aureus