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Literature summary for 4.1.2.13 extracted from
- One-step purification and characterization of a fully active histidine-tagged Class II fructose-1,6-bisphosphate aldolase from Mycobacterium tuberculosis (2008), Enzyme Microb. Technol., 43, 500-506.
No PubMed abstract available
Activating Compound
| Activating Compound | Comment | Organism | Structure |
|---|---|---|---|
| 2-mercaptoethanol | sulfhydryl compound such as 2-mercaptoethanol, activates the C-His-rMtFBA activity slightly, 15% | Mycobacterium tuberculosis |  |
| dithiothreitol | sulfhydryl compound such as dithiothreitol, activates the C-His-rMtFBA activity slightly, 25% | Mycobacterium tuberculosis | |
| additional information | 1 mM PMSF has no effect on the enzyme activity | Mycobacterium tuberculosis | |
| Triton X-100 | increased enzyme activity by 33% | Mycobacterium tuberculosis | |
Application
| Application | Comment | Organism |
|---|---|---|
| medicine | potential drug target | Mycobacterium tuberculosis |
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| expressed C-terminal histidine-tagged Class II FBA is produced in Escherichia coli BL21 | Mycobacterium tuberculosis |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| C-His-rMtFBA | C-terminal histidine-tagged Class II FBA | Mycobacterium tuberculosis |
General Stability
| General Stability | Organism |
|---|---|
| stored in the 50 mM Tris-HCl buffer, pH 7.4 containing 1 mM 2-mercaptoethanol and 100 microM ZnCl2, stable at 4°C for over 10 months without significant loss of activity. | Mycobacterium tuberculosis |
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| 2-propanol | at 12.5% (v/v) remaining activity, 69.99%, and 25% (v/v) remaining activity, 3.72% | Mycobacterium tuberculosis | |
| 5-chloro-8-hydroxyquinoline | non-competitive (mixed type) | Mycobacterium tuberculosis | |
| 8-hydroxyquinoline | - |
Mycobacterium tuberculosis | |
| Benzene | at 12.5% (v/v) remaining activity, 88.94%, and 25% (v/v) remaining activity, 88.94% | Mycobacterium tuberculosis | |
| dimethylsulfoxid | at 12.5% (v/v) remaining activity, 131.26%, and 25% (v/v) remaining activity, 133.2% | Mycobacterium tuberculosis | |
| EDTA | - |
Mycobacterium tuberculosis | |
| ethanol | at 12.5% (v/v) remaining activity, 79.19%, and 25% (v/v) remaining activity, 33.41% | Mycobacterium tuberculosis | |
| ethyl acetate | at 12.5% (v/v) remaining activity, 71.96%, and 25% (v/v) remaining activity, 41.62% | Mycobacterium tuberculosis | |
| glycerol | at 20% (v/v) reduced enzyme activity by 65% | Mycobacterium tuberculosis | |
| hexane | at 12.5% (v/v) remaining activity, 98.25%, and 25% (v/v) remaining activity, 92.22% | Mycobacterium tuberculosis | |
| additional information | all methyl 4-oxo-2-butenoates, 3-hydroxy-2-pyrrolones, 1,4-bezoxazines and compounds containing 4-quinolone fragment does not inhibit rMtFBA | Mycobacterium tuberculosis | |
| N,N-dimethylmethanamide | at 12.5% (v/v) remaining activity, 118.40%, and 25% (v/v) remaining activity, 73.93% | Mycobacterium tuberculosis | |
| Toluene | at 12.5% (v/v) remaining activity, 89.70%, and 25% (v/v) remaining activity, 91.02% | Mycobacterium tuberculosis | |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.01503 | - |
D-fructose-1,6-bisphosphate | purified C-His-rMtFBA, indicates that the extra five histidine residues in C-His-rMtFBA has negligible effects on the kinetic properties of the enzyme | Mycobacterium tuberculosis | |
| 0.01598 | - |
D-fructose-1,6-bisphosphate | purified native-rMtFBA | Mycobacterium tuberculosis | |
Metals/Ions
| Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|
| Co2+ | addition of only Co2+ and Zn2+ can recover the activity to higher than half of the original | Mycobacterium tuberculosis | |
| Cu2+ | addition of Cu2+, Fe2+, Ni2+, Mg2+, and Mn2+ leads to only a small level of recovery of activity | Mycobacterium tuberculosis | |
| Fe2+ | addition of Cu2+, Fe2+, Ni2+, Mg2+, and Mn2+ leads to only a small level of recovery of activity | Mycobacterium tuberculosis | |
| Mg2+ | addition of Cu2+, Fe2+, Ni2+, Mg2+, and Mn2+ leads to only a small level of recovery of activity | Mycobacterium tuberculosis | |
| Mn2+ | addition of Cu2+, Fe2+, Ni2+, Mg2+, and Mn2+ leads to only a small level of recovery of activity | Mycobacterium tuberculosis | |
| Ni2+ | addition of Cu2+, Fe2+, Ni2+, Mg2+, and Mn2+ leads to only a small level of recovery of activity | Mycobacterium tuberculosis | |
| Zn2+ | addition of only Co2+ and Zn2+ can recover the activity to higher than half of the original | Mycobacterium tuberculosis | |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| 37000 | - |
purified native-rMtFBA | Mycobacterium tuberculosis |
| 38000 | - |
purified C-His-rMtFBA | Mycobacterium tuberculosis |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Mycobacterium tuberculosis | P9WQA3 | - |
- |
| Mycobacterium tuberculosis H37Rv | P9WQA3 | - |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| immobilized metal affinity chromatography, Ni2+-IDA affinity chromatography | Mycobacterium tuberculosis |
Specific Activity [micromol/min/mg]
| Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|
| 34.85 | - |
purified native-rMtFBA | Mycobacterium tuberculosis |
| 35.05 | - |
purified C-His-rMtFBA | Mycobacterium tuberculosis |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| D-fructose 1,6-bisphosphate | - |
Mycobacterium tuberculosis | glycerone phosphate + D-glyceraldehyde 3-phosphate | - |
? | |
| D-fructose 1,6-bisphosphate | - |
Mycobacterium tuberculosis H37Rv | glycerone phosphate + D-glyceraldehyde 3-phosphate | - |
? | |
| additional information | similar to other Class II FBA, neither fructose 1-phosphate nor fructose 6-phosphate (up to 50 mM each) act as substrates for both rMtFBA | Mycobacterium tuberculosis | ? | - |
? | |
| additional information | similar to other Class II FBA, neither fructose 1-phosphate nor fructose 6-phosphate (up to 50 mM each) act as substrates for both rMtFBA | Mycobacterium tuberculosis H37Rv | ? | - |
? | |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| class II fructose-1,6-bisphosphate aldolase | - |
Mycobacterium tuberculosis |
| FBA | - |
Mycobacterium tuberculosis |
| MtFBA | - |
Mycobacterium tuberculosis |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 55 | - |
- |
Mycobacterium tuberculosis |
Temperature Stability [°C]
| Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 30 | 55 | activity lost substantially at higher temperature | Mycobacterium tuberculosis |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 7.8 | - |
- |
Mycobacterium tuberculosis |
pH Stability
| pH Stability | pH Stability Maximum | Comment | Organism |
|---|---|---|---|
| 7 | 10 | C-His-rMtFBA and native-rMtFBA are stable in a wider pH range of 7.0-10.0 | Mycobacterium tuberculosis |
Ki Value [mM]
| Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.1824 | - |
5-chloro-8-hydroxyquinoline | native-rMtFBA, Ki = 182.4 microM and KI = 219.5 microM | Mycobacterium tuberculosis | |
| 0.1934 | - |
5-chloro-8-hydroxyquinoline | C-His-rMtFBA, Ki = 193.4 microM and KI = 220.9 microM. This indicates a possible use of C-His-rMtFBA for screening of MtFBA inhibitors | Mycobacterium tuberculosis | |
pI Value
| Organism | Comment | pI Value Maximum | pI Value |
|---|---|---|---|
| Mycobacterium tuberculosis | native-rMtFBA, agreeing with the calculation from their sequence, 5.42 | - |
5.6 |
| Mycobacterium tuberculosis | C-His-rMtFBA, agreeing with the calculation from their sequence, 5.82 | - |
5.9 |
IC50 Value
| IC50 Value | IC50 Value Maximum | Comment | Organism | Inhibitor | Structure |
|---|---|---|---|---|---|
| 0.33 | - |
NADH-linked enzymatic assay | Mycobacterium tuberculosis | 5-chloro-8-hydroxyquinoline | |
| 0.496 | - |
NADH-linked enzymatic assay | Mycobacterium tuberculosis | 8-hydroxyquinoline | |
| 0.5 | - |
determined by the colorimetric assay | Mycobacterium tuberculosis | 8-hydroxyquinoline | |
| 0.533 | - |
determined by the colorimetric assay | Mycobacterium tuberculosis | 5-chloro-8-hydroxyquinoline | |
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