Cloned (Comment) | Organism |
---|---|
mutant HpaI enzymes expressed in Escherichia coli Bl21(lambda DE3) cells | Escherichia coli |
Protein Variants | Comment | Organism |
---|---|---|
H45A | by site-directed mutagenesis, mutant enzyme has 24fold higher dissociation constant for Co2+ compared to the wild-type enzyme, apparent Km value for Co2+ increases by about 800fold compared to the wild-type enzyme, different kcat and km values | Escherichia coli |
H45Q | by site-directed mutagenesis, mutant enzyme has 69fold higher dissociation constant for Co2+ compared to the wild-type enzyme, apparent Km value for Co2+ increases by about 800fold compared to the wild-type enzyme, different kcat and km values | Escherichia coli |
General Stability | Organism |
---|---|
the engineering results show that His45 has a structural and catalytic role. It is important for metal cofactor binding, possibly by proper positioning of a metal cofactor water ligand and is also involved in base catalysis. | Escherichia coli |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | apparent Km values for Co2+ increase in the two mutants H45A and H45Q by about 800fold compared to the wild-type enzyme | Escherichia coli | |
0.38 | - |
4-hydroxy-2-oxopentanoic acid | wild-type, pH 8.0, 25°C | Escherichia coli | |
16 | - |
4-hydroxy-2-oxopentanoic acid | H45Q mutant enzyme, pH 8.0, 25°C | Escherichia coli | |
38 | - |
4-hydroxy-2-oxopentanoic acid | H45A mutant enzyme, pH 8.0, 25°C | Escherichia coli |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
additional information | H45A and H45Q mutant enzymes have 24fold and 69fold higher dissociation constants for Co2+ compared to the wild-type enzyme | Escherichia coli |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
additional information | - |
sizes of the native mutant enzymes are identical to that of the wild-type enzyme as determined by gel filtration | Escherichia coli |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | Q47098 | - |
- |
Purification (Comment) | Organism |
---|---|
by anion exchange, hydrophobic interaction, and gel filtration chormatography | Escherichia coli |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
4-hydroxy-2-oxopentanoic acid | - |
Escherichia coli | pyruvate + ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
HpaI | - |
Escherichia coli |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.17 | - |
4-hydroxy-2-oxopentanoic acid | H45Q mutant enzyme, pH 8.0, 25°C | Escherichia coli | |
4.5 | - |
4-hydroxy-2-oxopentanoic acid | H45A mutant enzyme, pH 8.0, 25°C | Escherichia coli | |
350 | - |
4-hydroxy-2-oxopentanoic acid | wild-type, pH 8.0, 25°C | Escherichia coli |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
5 | 9 | - |
Escherichia coli |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.011 | - |
4-hydroxy-2-oxopentanoic acid | H45Q mutant enzyme, pH 8.0, 25°C | Escherichia coli | |
0.12 | - |
4-hydroxy-2-oxopentanoic acid | H45A mutant enzyme, pH 8.0, 25°C | Escherichia coli | |
930 | - |
4-hydroxy-2-oxopentanoic acid | wild-type, pH 8.0, 25°C | Escherichia coli |