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Literature summary 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

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.3.1.98 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

EC Number Protein Variants Comment Organism
1.3.1.98 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
1.3.1.98 G67A site-directed mutagenesis, functional complementation of murB-deficient, temperature-sensitive Staphylcoccus aureus mutant strain TS2901 Staphylococcus aureus
1.3.1.98 G69A site-directed mutagenesis, functional complementation of murB-deficient, temperature-sensitive Staphylcoccus aureus mutant strain TS2901 Staphylococcus aureus
1.3.1.98 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
1.3.1.98 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
1.3.1.98 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
1.3.1.98 R213A site-directed mutagenesis, nearly inactive mutant, no complementation of murB-deficient, temperature-sensitive Staphylcoccus aureus mutant strain TS2901 Staphylococcus aureus
1.3.1.98 R230A site-directed mutagenesis, functional complementation of murB-deficient, temperature-sensitive Staphylcoccus aureus mutant strain TS2901 Staphylococcus aureus
1.3.1.98 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
1.3.1.98 S70A site-directed mutagenesis, functional complementation of murB-deficient, temperature-sensitive Staphylcoccus aureus mutant strain TS2901 Staphylococcus aureus
1.3.1.98 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]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.3.1.98 additional information
-
 additional information kinetics of recombinant wild-type and mutant enzymes Staphylococcus aureus
1.3.1.98 0.0033
-
 NADPH pH 8.0, 22°C, recombinant mutant S226A Staphylococcus aureus
1.3.1.98 0.008
-
 NADPH pH 8.0, 22°C, recombinant mutant N71A Staphylococcus aureus
1.3.1.98 0.0095
-
 NADPH pH 8.0, 22°C, recombinant mutant Y175F Staphylococcus aureus
1.3.1.98 0.014
-
 NADPH pH 8.0, 22°C, recombinant mutant E296A Staphylococcus aureus
1.3.1.98 0.023
-
 NADPH pH 8.0, 22°C, recombinant mutant R176A Staphylococcus aureus
1.3.1.98 0.024
-
 NADPH pH 8.0, 22°C, recombinant mutant H259A Staphylococcus aureus
1.3.1.98 0.036
-
 NADPH pH 8.0, 22°C, recombinant mutant R213A Staphylococcus aureus
1.3.1.98 0.038
-
 NADPH pH 8.0, 22°C, recombinant wild-type enzyme Staphylococcus aureus

Natural Substrates/ Products (Substrates)

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

Organism

EC Number Organism UniProt Comment Textmining
1.3.1.98 Staphylococcus aureus P61431 gene murB
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.3.1.98 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

EC Number Reaction Comment Organism Reaction ID
1.3.1.98 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]

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

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.3.1.98 UDP-N-acetylenolpyruvylglucosamine + NADPH
-
 Staphylococcus aureus UDP-N-acetylmuramic acid + NADP+
-
 ?
1.3.1.98 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

EC Number Synonyms Comment Organism
1.3.1.98 MurB
-
 Staphylococcus aureus
1.3.1.98 UDP-N-acetylenolpyruvylglucosamine reductase
-
 Staphylococcus aureus

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.3.1.98 22
-
 assay at room temperature Staphylococcus aureus

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.3.1.98 8
-
 assay at Staphylococcus aureus

Cofactor

EC Number Cofactor Comment Organism Structure
1.3.1.98 FAD bound Staphylococcus aureus
1.3.1.98 NADPH dependent on Staphylococcus aureus