6.3.2.9: UDP-N-acetylmuramoyl-L-alanine-D-glutamate ligase
This is an abbreviated version!
For detailed information about UDP-N-acetylmuramoyl-L-alanine-D-glutamate ligase, go to the full flat file.
Reaction
Synonyms
D-Glutamate-adding enzyme, D-glutamic acid adding enzyme, D-Glutamic acid-adding enzyme, MurD, MurD cell wall enzyme, MurD ligase, Synthetase, uridine diphospho-N-acetylmuramoylalanyl-D-glutamate, UDP-Mur-Nac-L-Ala:L-Glu ligase, UDP-N-acetylmuramoyl-L-alanine:D-glutamate ligase, UDP-N-acetylmuramoyl-L-alanine:D-glutamateligase, UDP-N-acetylmuramoyl-L-alanyl-D-glutamate synthetase, UDPMurNAc-L-alanine:D-glutamate ligase (ADP-forming), Uridine diphosphate N-acetylmuramoyl-L-alanine:D-glutamate ligase, Uridine diphospho-N-acetylmuramoylalanyl-D-glutamate synthetase
ECTree
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Purification
Purification on EC 6.3.2.9 - UDP-N-acetylmuramoyl-L-alanine-D-glutamate ligase
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method that includes DEAE-Trisacryl M and hydroxyapatite-Ultrogel chromatography. Large-scale purification using 2-mercaptoethanol as a reducing agent, results in 200 mg of purified enzyme from 54 l of culture, adducts with one or two molecules of 2-mercaptoethanol are formed. Replacement of 2-mercaptoethanol by dithiothreitol gave 162 mg of enzyme from 36 l of culture yielding no clusters with the reducing agent.
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recombinant His6-tagged enzyme from Escherichia coli strain DH5alpha by nickel affinity chromatography and dialysis
recombinant recombinant GST-tagged MurD from Escherichia coli by glutathione affinity chromatography, His-tagged MurD from Mycobacterium smegmatis by nickel affinity chromatography