EC Number | Cloned (Comment) | Organism |
---|---|---|
5.1.1.1 | gene alr, recombinant His-tagged enzyme in Escherichia coli strain BL21(DE3) | Bacillus subtilis |
5.1.1.1 | gene alr, recombinant His-tagged enzyme in Escherichia coli strain BL21(DE3) | Mycobacterium tuberculosis |
5.1.1.3 | gene murI, recombinant His-tagged enzyme in Escherichia coli strain BL21(DE3) | Bacillus subtilis |
5.1.1.3 | gene murI, recombinant His-tagged enzyme in Escherichia coli strain BL21(DE3) | Mycobacterium tuberculosis |
EC Number | Inhibitors | Comment | Organism | Structure |
---|---|---|---|---|
5.1.1.1 | beta-Chloro-D-alanine | BCDA, its primary target is glutamate racemase, poor activity oagainst alanine racemase activity, potent antituberculosis activity. BCDA does not inhibit the D-alanine pathway in intact cells, consistent with its poor in vitro activity, it is instead an irreversible mechanism-based inactivator of glutamate racemase (MurI), an upstream enzyme in the same early stage of peptidoglycan biosynthesis. Inhibition kinetics, overview. Glutamate racemase (MurI) is a pyridoxal 5'-phosphate-independent racemase and is therefore unable to undergo the same mechanism of inhibition as Alr with BCDA | Bacillus subtilis | |
5.1.1.1 | beta-Chloro-D-alanine | BCDA, the compund is a very poor inhibitor of recombinant Mycobacterium tuberculosis Alr, despite having potent antituberculosis activity, its primary target is glutamate racemase, poor activity oagainst alanine racemase activity, potent antituberculosis activity. BCDA does not inhibit the D-alanine pathway in intact cells, consistent with its poor in vitro activity, it is instead an irreversible mechanism-based inactivator of glutamate racemase (MurI), an upstream enzyme in the same early stage of peptidoglycan biosynthesis. Inhibition kinetics, overview. Glutamate racemase (MurI) is a pyridoxal 5'-phosphate-independent racemase and is therefore unable to undergo the same mechanism of inhibition as Alr with BCDA | Mycobacterium tuberculosis | |
5.1.1.3 | beta-Chloro-D-alanine | BCDA, its primary target is glutamate racemase, poor activity oagainst alanine racemase activity, potent antituberculosis activity. BCDA does not inhibit the D-alanine pathway in intact cells, consistent with its poor in vitro activity, it is instead an irreversible mechanism-based inactivator of glutamate racemase (MurI), an upstream enzyme in the same early stage of peptidoglycan biosynthesis. Inhibition kinetics, overview. BCDA-treated BsMurI has a single cysteine residue, C185, that is the sole site of modification in over 95% of the inactivated protein. BCDA inhibition of MurI is mechanism-based as opposed to arising from nonspecific interaction with suitably configured cysteine thiols | Bacillus subtilis | |
5.1.1.3 | beta-Chloro-D-alanine | BCDA, its primary target is glutamate racemase, poor activity oagainst alanine racemase activity, potent antituberculosis activity. BCDA does not inhibit the D-alanine pathway in intact cells, consistent with its poor in vitro activity, it is instead an irreversible mechanism-based inactivator of glutamate racemase (MurI), an upstream enzyme in the same early stage of peptidoglycan biosynthesis. Inhibition kinetics, overview. BCDA modifies MtMurI in vitro and in vivo | Mycobacterium tuberculosis | |
5.1.1.3 | beta-chloro-L-alanine | BCLA, modifies BsMurI predominantly at C74, the cysteine residue responsible for deprotonation of an incoming D-Glu substrate | Bacillus subtilis | |
5.1.1.3 | additional information | no inhibition by beta-fluoroalanine (BFA, racemic mixture) and O-acetyl-D-serine (OADS) | Bacillus subtilis | |
5.1.1.3 | additional information | no inhibition by beta-fluoroalanine (BFA, racemic mixture) and O-acetyl-D-serine (OADS). MtMurI adopts a unique oligomeric configuration and contains distinct active-site architectural differences from other MurI orthologues and that dimer interface mutations are required to introduce catalytic capability. Conventional MurI inhibitors are inactive against this recombinant form of mycobacterial MurI, the selective inhibitory activity of BCDA against MtMurI may arise from unique protein-ligand interactions unseen in orthologous MurIs. MsMurI results demonstrate these same structural features and yet BCDA does not appear to act via MurI in this bacterium | Mycobacterium tuberculosis |
EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
5.1.1.1 | 3.7 | - |
L-alanine | pH 7.6, 37°C | Mycobacterium tuberculosis | |
5.1.1.1 | 5.9 | - |
L-alanine | pH 7.6, 37°C | Bacillus subtilis | |
5.1.1.3 | 1.5 | - |
D-glutamate | pH 7.6, 37°C | Mycobacterium tuberculosis | |
5.1.1.3 | 37 | - |
L-glutamate | pH 7.6, 37°C | Mycobacterium tuberculosis |
EC Number | Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
5.1.1.1 | L-alanine | Bacillus subtilis | - |
D-alanine | - |
r | |
5.1.1.1 | L-alanine | Mycobacterium tuberculosis | - |
D-alanine | - |
r | |
5.1.1.1 | L-alanine | Bacillus subtilis 168 | - |
D-alanine | - |
r | |
5.1.1.1 | L-alanine | Mycobacterium tuberculosis ATCC 25618 / H37Rv | - |
D-alanine | - |
r | |
5.1.1.3 | L-glutamate | Bacillus subtilis | - |
D-glutamate | - |
r | |
5.1.1.3 | L-glutamate | Mycobacterium tuberculosis | - |
D-glutamate | - |
r | |
5.1.1.3 | L-glutamate | Bacillus subtilis 168 | - |
D-glutamate | - |
r | |
5.1.1.3 | L-glutamate | Mycobacterium tuberculosis ATCC 25618 / H37Rv | - |
D-glutamate | - |
r |
EC Number | Organism | UniProt | Comment | Textmining |
---|---|---|---|---|
5.1.1.1 | Bacillus subtilis | P94556 | - |
- |
5.1.1.1 | Bacillus subtilis 168 | P94556 | - |
- |
5.1.1.1 | Mycobacterium tuberculosis | P9WPW9 | - |
- |
5.1.1.1 | Mycobacterium tuberculosis ATCC 25618 / H37Rv | P9WPW9 | - |
- |
5.1.1.3 | Bacillus subtilis | P94556 | - |
- |
5.1.1.3 | Bacillus subtilis 168 | P94556 | - |
- |
5.1.1.3 | Mycobacterium tuberculosis | P9WPW9 | - |
- |
5.1.1.3 | Mycobacterium tuberculosis ATCC 25618 / H37Rv | P9WPW9 | - |
- |
EC Number | Purification (Comment) | Organism |
---|---|---|
5.1.1.1 | recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, tag cleavage | Bacillus subtilis |
5.1.1.1 | recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, tag cleavage | Mycobacterium tuberculosis |
5.1.1.3 | recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, tag cleavage | Bacillus subtilis |
5.1.1.3 | recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, tag cleavage | Mycobacterium tuberculosis |
EC Number | Reaction | Comment | Organism | Reaction ID |
---|---|---|---|---|
5.1.1.3 | L-glutamate = D-glutamate | MurI is a member of the two-base mechanism (dual-cysteine) racemase family, where two essential active-site cysteine residues act as catalytic base and acid to stereospecifically de- and reprotonate, respectively, the alpha position of glutamate in order to enact substrate racemization | Mycobacterium tuberculosis | |
5.1.1.3 | L-glutamate = D-glutamate | MurI is a member of the two-base mechanism (dual-cysteine) racemase family, where two essential active-site cysteine residues act as catalytic base and acid to stereospecifically de- and reprotonate, respectively, the alpha position of glutamate in order to enact substrate racemization. C185 of BsMurI corresponds to the essential catalytic cysteine residue that deprotonates an incoming L-glutamate substrate (or reprotonates a carbanionic intermediate to form the D-stereoconfiguration) | Bacillus subtilis |
EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
5.1.1.1 | L-alanine | - |
Bacillus subtilis | D-alanine | - |
r | |
5.1.1.1 | L-alanine | - |
Mycobacterium tuberculosis | D-alanine | - |
r | |
5.1.1.1 | L-alanine | - |
Bacillus subtilis 168 | D-alanine | - |
r | |
5.1.1.1 | L-alanine | - |
Mycobacterium tuberculosis ATCC 25618 / H37Rv | D-alanine | - |
r | |
5.1.1.3 | D-glutamate | - |
Mycobacterium tuberculosis | L-glutamate | - |
r | |
5.1.1.3 | D-glutamate | - |
Mycobacterium tuberculosis ATCC 25618 / H37Rv | L-glutamate | - |
r | |
5.1.1.3 | L-glutamate | - |
Bacillus subtilis | D-glutamate | - |
r | |
5.1.1.3 | L-glutamate | - |
Mycobacterium tuberculosis | D-glutamate | - |
r | |
5.1.1.3 | L-glutamate | - |
Bacillus subtilis 168 | D-glutamate | - |
r | |
5.1.1.3 | L-glutamate | - |
Mycobacterium tuberculosis ATCC 25618 / H37Rv | D-glutamate | - |
r |
EC Number | Synonyms | Comment | Organism |
---|---|---|---|
5.1.1.1 | MurI | - |
Bacillus subtilis |
5.1.1.1 | MurI | - |
Mycobacterium tuberculosis |
5.1.1.3 | MurI | - |
Bacillus subtilis |
5.1.1.3 | MurI | - |
Mycobacterium tuberculosis |
EC Number | Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|---|
5.1.1.1 | 37 | - |
assay at | Bacillus subtilis |
5.1.1.1 | 37 | - |
assay at | Mycobacterium tuberculosis |
5.1.1.3 | 37 | - |
assay at | Bacillus subtilis |
5.1.1.3 | 37 | - |
assay at | Mycobacterium tuberculosis |
EC Number | Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
5.1.1.1 | 37 | - |
L-alanine | pH 7.6, 37°C | Mycobacterium tuberculosis | |
5.1.1.1 | 1190 | - |
L-alanine | pH 7.6, 37°C | Bacillus subtilis | |
5.1.1.3 | 12.6 | - |
D-glutamate | pH 7.6, 37°C | Mycobacterium tuberculosis | |
5.1.1.3 | 226.4 | - |
L-glutamate | pH 7.6, 37°C | Mycobacterium tuberculosis |
EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|---|
5.1.1.1 | 7.6 | - |
assay at | Bacillus subtilis |
5.1.1.1 | 7.6 | - |
assay at | Mycobacterium tuberculosis |
5.1.1.3 | 7.6 | - |
assay at | Bacillus subtilis |
5.1.1.3 | 7.6 | - |
assay at | Mycobacterium tuberculosis |
EC Number | Cofactor | Comment | Organism | Structure |
---|---|---|---|---|
5.1.1.1 | pyridoxal 5'-phosphate | - |
Mycobacterium tuberculosis | |
5.1.1.3 | additional information | glutamate racemase (MurI) is a pyridoxal 5'-phosphate-independent racemase | Bacillus subtilis | |
5.1.1.3 | additional information | glutamate racemase (MurI) is a pyridoxal 5'-phosphate-independent racemase | Mycobacterium tuberculosis |
EC Number | Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|---|
5.1.1.1 | 0.13 | - |
beta-Chloro-D-alanine | pH 7.6, 37°C | Mycobacterium tuberculosis | |
5.1.1.1 | 0.23 | - |
beta-Chloro-D-alanine | pH 7.6, 37°C | Bacillus subtilis | |
5.1.1.3 | 0.13 | - |
beta-Chloro-D-alanine | pH 7.6, 37°C | Mycobacterium tuberculosis | |
5.1.1.3 | 0.23 | - |
beta-Chloro-D-alanine | pH 7.6, 37°C | Bacillus subtilis |
EC Number | kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
5.1.1.1 | 10 | - |
L-alanine | pH 7.6, 37°C | Mycobacterium tuberculosis | |
5.1.1.1 | 201.7 | - |
L-alanine | pH 7.6, 37°C | Bacillus subtilis | |
5.1.1.3 | 6.12 | - |
L-glutamate | pH 7.6, 37°C | Mycobacterium tuberculosis | |
5.1.1.3 | 8.4 | - |
D-glutamate | pH 7.6, 37°C | Mycobacterium tuberculosis |