Application | Comment | Organism |
---|---|---|
drug development | acetohydroxyacid synthase (AHAS) of Mycobacterium tuberculosis is a promising target for the development of anti-tuberculosis agents | Mycobacterium tuberculosis |
Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli BL21(DE3) cells | Mycobacterium tuberculosis |
sequence comparisoons, recombinant expression of soluble His-tagged enzyme mutants in Escherichia coli strain BL21(DE3) | Mycobacterium tuberculosis |
Protein Variants | Comment | Organism |
---|---|---|
P126A | site-directed mutagenesis, the mutant exhibits similar kinetics but significantly lower activity compared to the wild-type enzyme | Mycobacterium tuberculosis |
P126A, | the mutant exhibits significantly lower activity than the wild type enzyme | Mycobacterium tuberculosis |
P126E | inactive | Mycobacterium tuberculosis |
P126E | site-directed mutagenesis, inactive mutant | Mycobacterium tuberculosis |
P126T | site-directed mutagenesis, the mutant exhibits significantly lower activity than wild-type enzyme and a significantly decreased preference toward thiamine diphosphate as cofactor | Mycobacterium tuberculosis |
P126T | the mutant exhibits significantly lower activity than the wild type enzyme | Mycobacterium tuberculosis |
P126V | site-directed mutagenesis, the mutant exhibits significantly lower activity than wild-type enzyme and a significantly decreased preference toward pyruvate as substrate | Mycobacterium tuberculosis |
P126V | the mutant exhibits significantly lower activity than the wild type enzyme | Mycobacterium tuberculosis |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
chlorimuron ethyl | a sulfonylurea derivative herbicide | Mycobacterium tuberculosis |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
2.76 | - |
pyruvate | pH 7.5, 37°C, recombinant wild-type enzyme | Mycobacterium tuberculosis | |
2.76 | - |
pyruvate | wild type enzyme, at pH 7.5 and 37°C | Mycobacterium tuberculosis | |
6.04 | - |
pyruvate | pH 7.5, 37°c, recombinant mutant P126A | Mycobacterium tuberculosis | |
6.04 | - |
pyruvate | mutant enzyme P126A, at pH 7.5 and 37°C | Mycobacterium tuberculosis | |
466.7 | - |
pyruvate | pH 7.5, 37°c, recombinant mutant P126V | Mycobacterium tuberculosis | |
466.7 | - |
pyruvate | mutant enzyme P126V, at pH 7.5 and 37°C | Mycobacterium tuberculosis | |
807.7 | - |
pyruvate | pH 7.5, 37°c, recombinant mutant P126T | Mycobacterium tuberculosis | |
807.7 | - |
pyruvate | mutant enzyme P126T, at pH 7.5 and 37°C | Mycobacterium tuberculosis |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | required | Mycobacterium tuberculosis | |
Mg2+ | the enzyme requires a divalent metal ion such as Mg2+ to anchor thiamine diphosphate | Mycobacterium tuberculosis |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
68000 | - |
x * 68000, SDS-PAGE | Mycobacterium tuberculosis |
68000 | - |
x * 68000, about, wild-type and mutant enzyme variants, SDS-PAGE | Mycobacterium tuberculosis |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
2 pyruvate | Mycobacterium tuberculosis | - |
2-acetolactate + CO2 | - |
ir | |
2 pyruvate | Mycobacterium tuberculosis | - |
2-acetolactate + CO2 | - |
? | |
2 pyruvate | Mycobacterium tuberculosis H37Rv | - |
2-acetolactate + CO2 | - |
ir | |
2 pyruvate | Mycobacterium tuberculosis H37Rv | - |
2-acetolactate + CO2 | - |
? | |
pyruvate + 2-oxobutyrate | Mycobacterium tuberculosis | - |
2-aceto-2-hydroxybutyrate + CO2 | - |
ir | |
pyruvate + 2-oxobutyrate | Mycobacterium tuberculosis H37Rv | - |
2-aceto-2-hydroxybutyrate + CO2 | - |
ir |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Mycobacterium tuberculosis | - |
- |
- |
Mycobacterium tuberculosis | P9WG39 | - |
- |
Mycobacterium tuberculosis H37Rv | - |
- |
- |
Mycobacterium tuberculosis H37Rv | P9WG39 | - |
- |
Purification (Comment) | Organism |
---|---|
- |
Mycobacterium tuberculosis |
recombinant soluble His-tagged enzyme mutants from Escherichia coli strain BL21(DE3) to near homogeneity by nickel affinity chromatography, gel filtration, and anion exchange chromatography | Mycobacterium tuberculosis |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
2 pyruvate | - |
Mycobacterium tuberculosis | 2-acetolactate + CO2 | - |
ir | |
2 pyruvate | - |
Mycobacterium tuberculosis | 2-acetolactate + CO2 | - |
? | |
2 pyruvate | - |
Mycobacterium tuberculosis H37Rv | 2-acetolactate + CO2 | - |
ir | |
2 pyruvate | - |
Mycobacterium tuberculosis H37Rv | 2-acetolactate + CO2 | - |
? | |
pyruvate + 2-oxobutyrate | - |
Mycobacterium tuberculosis | 2-aceto-2-hydroxybutyrate + CO2 | - |
ir | |
pyruvate + 2-oxobutyrate | - |
Mycobacterium tuberculosis H37Rv | 2-aceto-2-hydroxybutyrate + CO2 | - |
ir |
Subunits | Comment | Organism |
---|---|---|
? | x * 68000, SDS-PAGE | Mycobacterium tuberculosis |
? | x * 68000, about, wild-type and mutant enzyme variants, SDS-PAGE | Mycobacterium tuberculosis |
Synonyms | Comment | Organism |
---|---|---|
acetohydroxyacid synthase | - |
Mycobacterium tuberculosis |
AHAS | - |
Mycobacterium tuberculosis |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Mycobacterium tuberculosis |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.164 | - |
pyruvate | pH 7.5, 37°c, recombinant mutant P126V | Mycobacterium tuberculosis | |
0.164 | - |
pyruvate | mutant enzyme P126V, at pH 7.5 and 37°C | Mycobacterium tuberculosis | |
0.165 | - |
pyruvate | pH 7.5, 37°c, recombinant mutant P126A | Mycobacterium tuberculosis | |
0.165 | - |
pyruvate | mutant enzyme P126A, at pH 7.5 and 37°C | Mycobacterium tuberculosis | |
0.166 | - |
pyruvate | pH 7.5, 37°c, recombinant mutant P126T | Mycobacterium tuberculosis | |
0.166 | - |
pyruvate | mutant enzyme P126T, at pH 7.5 and 37°C | Mycobacterium tuberculosis | |
3.196 | - |
pyruvate | pH 7.5, 37°C, recombinant wild-type enzyme | Mycobacterium tuberculosis | |
3.196 | - |
pyruvate | wild type enzyme, at pH 7.5 and 37°C | Mycobacterium tuberculosis |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
assay at | Mycobacterium tuberculosis |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
FAD | dependent on | Mycobacterium tuberculosis | |
thiamine diphosphate | - |
Mycobacterium tuberculosis | |
thiamine diphosphate | dependent on, the cofactor plays a key role in catalysis. The thiamine diphosphate binding pocket contains the highly conserved proline 126 residue, binding pocket structure, overview. Thiamine diphosphate is located centrally in the active site of AHAS with a unique V-conformation at the dimer interface. In the dimeric structure, one subunit is in contact with the diphosphate moiety of thiamine diphosphate, and the other subunit is in contact with the aminopyrimidine moiety | Mycobacterium tuberculosis |
General Information | Comment | Organism |
---|---|---|
metabolism | the enzyme is involved in the branched chain amino acid biosynthesis | Mycobacterium tuberculosis |
additional information | structure homology modeling | Mycobacterium tuberculosis |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.205 | - |
pyruvate | pH 7.5, 37°c, recombinant mutant P126T | Mycobacterium tuberculosis | |
0.352 | - |
pyruvate | pH 7.5, 37°c, recombinant mutant P126V | Mycobacterium tuberculosis | |
27.22 | - |
pyruvate | pH 7.5, 37°c, recombinant mutant P126A | Mycobacterium tuberculosis | |
1158 | - |
pyruvate | pH 7.5, 37°C, recombinant wild-type enzyme | Mycobacterium tuberculosis |