Cloned (Comment) | Organism |
---|---|
gene mca or NCgl0948, quantitative real-time PCR enzyme expression analysis, recombinant wild-type and mutant enzyme expression in Escherichia coli strain BL21(DE3) | Corynebacterium glutamicum |
Protein Variants | Comment | Organism |
---|---|---|
D132A | site-directed mutagenesis, the mutant shows slightly decreased activity compared to the wild-type enzyme | Corynebacterium glutamicum |
D141A | site-directed mutagenesis, the mutant shows decreased activity compared to the wild-type enzyme | Corynebacterium glutamicum |
D14A | site-directed mutagenesis, the mutant shows slightly decreased activity compared to the wild-type enzyme | Corynebacterium glutamicum |
D15A | site-directed mutagenesis, the mutant shows increased activity compared to the wild-type enzyme | Corynebacterium glutamicum |
E16A | site-directed mutagenesis, the mutant shows slightly decreased activity compared to the wild-type enzyme | Corynebacterium glutamicum |
E43A | site-directed mutagenesis, the mutant shows increased activity compared to the wild-type enzyme | Corynebacterium glutamicum |
H10A | site-directed mutagenesis, the mutant shows decreased activity compared to the wild-type enzyme | Corynebacterium glutamicum |
H12A | site-directed mutagenesis, the mutant shows decreased activity compared to the wild-type enzyme | Corynebacterium glutamicum |
H139A | site-directed mutagenesis, the mutant shows decreased activity compared to the wild-type enzyme | Corynebacterium glutamicum |
H142A | site-directed mutagenesis, the mutant shows slightly decreased activity compared to the wild-type enzyme | Corynebacterium glutamicum |
additional information | construction of a mca deletion mutant | Corynebacterium glutamicum |
Y137A | site-directed mutagenesis, the mutant shows decreased activity compared to the wild-type enzyme | Corynebacterium glutamicum |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
Cd2+ | - |
Corynebacterium glutamicum | |
Cr2+ | - |
Corynebacterium glutamicum | |
Cu2+ | - |
Corynebacterium glutamicum | |
additional information | heavy metal ions, including Cd2+, Ni2+, Cr2+ and Cu2+, markedly inhibit growth of the mca mutant relative to the wild type strain | Corynebacterium glutamicum | |
Ni2+ | - |
Corynebacterium glutamicum |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | Michaelis-Menten kinetics of wild-type and mutant enzymes with substrate N-acetyl-D-glucosamine, overview | Corynebacterium glutamicum | |
92.34 | - |
mycothiol bimane | pH 7.5, 30°C, recombinant wild-type enzyme | Corynebacterium glutamicum | |
93.37 | - |
mycothiol bimane | pH 7.5, 30°C, recombinant mutant E43A | Corynebacterium glutamicum | |
99.92 | - |
mycothiol bimane | pH 7.5, 30°C, recombinant mutant D132A | Corynebacterium glutamicum | |
107.5 | - |
mycothiol bimane | pH 7.5, 30°C, recombinant mutant E16A | Corynebacterium glutamicum | |
108.4 | - |
mycothiol bimane | pH 7.5, 30°C, recombinant mutant H10A | Corynebacterium glutamicum | |
111.6 | - |
mycothiol bimane | pH 7.5, 30°C, recombinant mutant H12A | Corynebacterium glutamicum | |
115.3 | - |
mycothiol bimane | pH 7.5, 30°C, recombinant mutant H142A | Corynebacterium glutamicum | |
128.8 | - |
mycothiol bimane | pH 7.5, 30°C, recombinant mutant D15A | Corynebacterium glutamicum | |
132.3 | - |
mycothiol bimane | pH 7.5, 30°C, recombinant mutant D14A | Corynebacterium glutamicum | |
138 | - |
mycothiol bimane | pH 7.5, 30°C, recombinant mutant D141A | Corynebacterium glutamicum | |
469.6 | - |
mycothiol bimane | pH 7.5, 30°C, recombinant mutant H139A | Corynebacterium glutamicum | |
542.4 | - |
mycothiol bimane | pH 7.5, 30°C, recombinant mutant Y137A | Corynebacterium glutamicum |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Zn2+ | metal ion requirement, activates | Corynebacterium glutamicum |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
35000 | - |
recombinant enzyme, gel filtration | Corynebacterium glutamicum |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | Corynebacterium glutamicum | the wild-type enzyme is active on a mycothiol S-conjugate of monobromobimane (MSmB) in vivo | ? | - |
? | |
additional information | Corynebacterium glutamicum DSM 20300 | the wild-type enzyme is active on a mycothiol S-conjugate of monobromobimane (MSmB) in vivo | ? | - |
? | |
mycothiol bimane + H2O | Corynebacterium glutamicum | - |
mycothiol + bimane | - |
? | |
mycothiol bimane + H2O | Corynebacterium glutamicum DSM 20300 | - |
mycothiol + bimane | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Corynebacterium glutamicum | Q8NRQ7 | RES167, gene mca or NCgl0948 | - |
Corynebacterium glutamicum DSM 20300 | Q8NRQ7 | RES167, gene mca or NCgl0948 | - |
Purification (Comment) | Organism |
---|---|
recombinant wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) to homogeneity | Corynebacterium glutamicum |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | the wild-type enzyme is active on a mycothiol S-conjugate of monobromobimane (MSmB) in vivo | Corynebacterium glutamicum | ? | - |
? | |
additional information | substrate specificity, overview. The enzyme also deacetylates N-acetyl-D-glucosamine | Corynebacterium glutamicum | ? | - |
? | |
additional information | the wild-type enzyme is active on a mycothiol S-conjugate of monobromobimane (MSmB) in vivo | Corynebacterium glutamicum DSM 20300 | ? | - |
? | |
additional information | substrate specificity, overview. The enzyme also deacetylates N-acetyl-D-glucosamine | Corynebacterium glutamicum DSM 20300 | ? | - |
? | |
mycothiol bimane + H2O | - |
Corynebacterium glutamicum | mycothiol + bimane | - |
? | |
mycothiol bimane + H2O | - |
Corynebacterium glutamicum DSM 20300 | mycothiol + bimane | - |
? |
Subunits | Comment | Organism |
---|---|---|
monomer | 1 * 35000, recombinant enzyme, SDS-PAGE | Corynebacterium glutamicum |
Synonyms | Comment | Organism |
---|---|---|
MCA | - |
Corynebacterium glutamicum |
mycothiol S-conjugate amidase | - |
Corynebacterium glutamicum |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
assay at | Corynebacterium glutamicum |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0048 | - |
mycothiol bimane | pH 7.5, 30°C, recombinant mutant H139A | Corynebacterium glutamicum | |
0.005 | - |
mycothiol bimane | pH 7.5, 30°C, recombinant mutant Y137A | Corynebacterium glutamicum | |
0.036 | - |
mycothiol bimane | pH 7.5, 30°C, recombinant mutant D14A | Corynebacterium glutamicum | |
0.046 | - |
mycothiol bimane | pH 7.5, 30°C, recombinant mutant D141A | Corynebacterium glutamicum | |
0.058 | - |
mycothiol bimane | pH 7.5, 30°C, recombinant mutant E43A | Corynebacterium glutamicum | |
0.059 | - |
mycothiol bimane | pH 7.5, 30°C, recombinant wild-type enzyme | Corynebacterium glutamicum | |
0.065 | - |
mycothiol bimane | pH 7.5, 30°C, recombinant mutant D132A | Corynebacterium glutamicum | |
0.0655 | - |
mycothiol bimane | pH 7.5, 30°C, recombinant mutant H142A | Corynebacterium glutamicum | |
0.066 | - |
mycothiol bimane | pH 7.5, 30°C, recombinant mutant H12A | Corynebacterium glutamicum | |
0.068 | - |
mycothiol bimane | pH 7.5, 30°C, recombinant mutant H10A | Corynebacterium glutamicum | |
0.069 | - |
mycothiol bimane | pH 7.5, 30°C, recombinant mutant E16A | Corynebacterium glutamicum | |
0.078 | - |
mycothiol bimane | pH 7.5, 30°C, recombinant mutant D15A | Corynebacterium glutamicum |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | 8.5 | recombinant enzyme | Corynebacterium glutamicum |
Organism | Comment | Expression |
---|---|---|
Corynebacterium glutamicum | induced expression of Mca in Corynebacterium glutamicum under various stress conditions, directly under the control of the stress-responsive extracytoplasmic function-sigma (ECF-s) factor SigH, positive regulation of mca expression by SigH | up |
General Information | Comment | Organism |
---|---|---|
evolution | some Gram-positive bacteria, such as members of Corynebacterium, Mycobacterium, Rhodococcus and Streptomyces, cannot produce GSH but instead synthesize its functional equivalent, mycothiol | Corynebacterium glutamicum |
malfunction | mutants lacking the enzyme and mycothiol are more susceptible to alkylating agents and oxidants, e.g. monobromobimane, iodoacetamide, N-ethylmaleimide, 1-chloro-2,4-dinitrobenzene, and methyllglyoxal, or to some macrolides and beta-lactams, e.g. rifamycin S, than the wild-type enzyme. Heavy metal ions, including Cd2+, Ni2+, Cr2+ and Cu2+, markedly inhibit growth of the mca mutant relative to the wild type strain | Corynebacterium glutamicum |
additional information | residues Asp14, Tyr137, His139 and Asp141 are important for enzyme activity | Corynebacterium glutamicum |
physiological function | mycothiol S-conjugate amidase is a key enzyme involved in MSH-dependent detoxification. In detoxification process, mycothiol directly reacts with electrophilic compounds by its thiol moiety, forming MSH S-conjugates, regulation mechanism, potential roles of the enzyme in resistance to alkylating agents and oxidants and in the survival of Corynebacterium glutamicum by coping with multiple stresses and detoxifying toxins, overview | Corynebacterium glutamicum |