Cloned (Comment) | Organism |
---|---|
gene murB, expression of His-tagged enzyme in Escherichia coli K12S | Staphylococcus aureus |
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 | Organism |
---|---|
repeated freeze-thaw cycles inactivate the purified recombinant His-tagged enzyme | Staphylococcus aureus |
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+ | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Staphylococcus aureus | - |
- |
- |
Staphylococcus aureus | - |
gene murB | - |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged enzyme from Escherichia coli K12S by nickel affinity chromatography | Staphylococcus aureus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
UDP-N-acetylenolpyruvylglucosamine + NADPH | - |
Staphylococcus aureus | UDP-N-acetylmuramic acid + NADP+ | - |
? | |
UDP-N-acetylenolpyruvylglucosamine + NADPH | biosynthetic MurB pathway, overview | Staphylococcus aureus | UDP-N-acetylmuramic acid + NADP+ | - |
? |
Synonyms | Comment | Organism |
---|---|---|
MurB | - |
Staphylococcus aureus |
UDP-N-acetylenolpyruvylglucosamine reductase | - |
Staphylococcus aureus |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
4 | - |
the purified recombinant His-tagged enzyme is unstable at 4°C | Staphylococcus aureus |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NADPH | - |
Staphylococcus aureus |