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Literature summary for 1.3.1.98 extracted from

  • Harris, M.S.; Herberg, J.T.; Cialdella, J.I.; Martin, J.T., Jr.; Benson, T.E.; Choi, G.H.; Baldwin, E.T.
    Crystallisation and preliminary X-ray analysis of UDP-N-acetylenolpyruvylglucosamine reductase (MurB) from Staphylococcus aureus (2001), Acta Crystallogr. Sect. D, 57, 1032-1035.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
gene murB, expression of His-tagged enzyme in Escherichia coli K12S Staphylococcus aureus

Crystallization (Commentary)

Crystallization (Comment) Organism
by sitting-drop vapour diffusion Staphylococcus aureus
purified recombinant His-tagged enzyme, 20 mg/ml protein in 20 mM HEPES, pH 7.5 5 mM 2-mercaptoethanol or DTT, 20°C, hanging drop vapour diffusion method, mixing with reservoir solution containing 0.1 M sodium cacodylate, pH 6.5, 15% PEG 8000, and 0.5 M (NH4)2SO4, 4 days, yellow plate-shaped crystals cryoprotected in 10% PEG 8000, 0.55 M (NH4)2SO4, 0.1 M sodium cacodylate, pH 6.5, 5 mM BME or DTT with 15% 2-methyl 2,4-pentanediol, X-ray diffraction structure determination and analysis at 2.3 A resolution, altered crystallization conditions were optimized using the purified selenomethionine-labeled MurB at 20 mg/ml, in complex with 10 mM enolpyruvyl-UDP-N-acetylglucosamine, in 1-2 weeks on grease-coated microbridges by mixing 5 ml Se-Met MurB and 5 ml reservoir solution containing 0.1 M sodium cacodylate, 9.75% PEG 8000, 0.55 M (NH4)2SO4, 20% DMSO, and 5 mM 2-mercaptoethanol and producing larger crystals, overview Staphylococcus aureus

General Stability

General Stability Organism
repeated freeze-thaw cycles inactivate the purified recombinant His-tagged 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 biosynthetic MurB pathway, overview UDP-N-acetylmuramic acid + NADP+
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Organism

Organism UniProt Comment Textmining
Staphylococcus aureus
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-
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Staphylococcus aureus
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gene murB
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Purification (Commentary)

Purification (Comment) Organism
recombinant His-tagged enzyme from Escherichia coli K12S by nickel affinity chromatography Staphylococcus aureus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
UDP-N-acetylenolpyruvylglucosamine + NADPH
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Staphylococcus aureus UDP-N-acetylmuramic acid + NADP+
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?
UDP-N-acetylenolpyruvylglucosamine + NADPH biosynthetic MurB pathway, overview Staphylococcus aureus UDP-N-acetylmuramic acid + NADP+
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?

Synonyms

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

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
4
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the purified recombinant His-tagged enzyme is unstable at 4°C Staphylococcus aureus

Cofactor

Cofactor Comment Organism Structure
NADPH
-
Staphylococcus aureus