Cloned (Comment) | Organism |
---|---|
gene leuB, expression of wild-type and mutant enzymes in Escherichia coli strain MA153 | Sulfurisphaera tokodaii |
Protein Variants | Comment | Organism |
---|---|---|
F145S | random mutagenesis, the mutation alters the Km values for NAD+ and/or the kcat value, the mutant shows reduced thermostability compared to the wild-type enzyme | Sulfurisphaera tokodaii |
F3S | random mutagenesis, the mutation alters the Km values for NAD+ and/or the kcat value, the mutant shows reduced thermostability compared to the wild-type enzyme | Sulfurisphaera tokodaii |
F3S/L288P | random mutagenesis, the mutation alters the Km values for NAD+ and/or the kcat value, the mutant shows reduced thermostability compared to the wild-type enzyme. The MA153 strain containing a doubly mutated leuB gene grows more rapidly than does Escherichia coli MA153 containing the wild-type leuB gene in leucine-free M9 minimal liquid medium | Sulfurisphaera tokodaii |
F3S/S232G | random mutagenesis, the mutation alters the Km values for NAD+ and/or the kcat value, the mutant shows reduced thermostability compared to the wild-type enzyme. The MA153 strain containing a doubly mutated leuB gene grows more rapidly than does Escherichia coli MA153 containing the wild-type leuB gene in leucine-free M9 minimal liquid medium | Sulfurisphaera tokodaii |
I68L/I142T/K335E | random mutagenesis, the mutation alters the Km values for NAD+ and/or the kcat value, the mutant shows reduced thermostability compared to the wild-type enzyme | Sulfurisphaera tokodaii |
L288P | random mutagenesis, the mutation alters the Km values for NAD+ and/or the kcat value, the mutant shows reduced thermostability compared to the wild-type enzyme | Sulfurisphaera tokodaii |
L328S | random mutagenesis, the mutation alters the Km values for NAD+ and/or the kcat value, the mutant shows reduced thermostability compared to the wild-type enzyme | Sulfurisphaera tokodaii |
L336P | random mutagenesis, the mutation alters the Km values for NAD+ and/or the kcat value, the mutant shows reduced thermostability compared to the wild-type enzyme | Sulfurisphaera tokodaii |
R284G | random mutagenesis, the mutation alters the Km values for NAD+ and/or the kcat value, the mutant shows reduced thermostability compared to the wild-type enzyme | Sulfurisphaera tokodaii |
Y309H | random mutagenesis, the mutation alters the Km values for NAD+ and/or the kcat value, the mutant shows reduced thermostability compared to the wild-type enzyme | Sulfurisphaera tokodaii |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | Michaelis-Menten kinetics of recombinant wild-type and mutant enzymes at different temperatures, overview | Sulfurisphaera tokodaii | |
0.0039 | - |
3-isopropylmalate | pH 7.5, 70°C, recombinant mutant F3S/S232G | Sulfurisphaera tokodaii | |
0.0044 | - |
3-isopropylmalate | pH 7.5, 70°C, recombinant mutant L288P | Sulfurisphaera tokodaii | |
0.0055 | - |
3-isopropylmalate | pH 7.5, 70°C, recombinant wild-type enzyme | Sulfurisphaera tokodaii | |
0.0067 | - |
3-isopropylmalate | pH 7.5, 70°C, recombinant mutant F3S/L288P | Sulfurisphaera tokodaii | |
0.0076 | - |
3-isopropylmalate | pH 7.5, 70°C, recombinant mutant F3S | Sulfurisphaera tokodaii | |
0.008 | - |
3-isopropylmalate | pH 7.5, 70°C, recombinant mutant R284G | Sulfurisphaera tokodaii | |
0.22 | - |
NAD+ | pH 7.5, 70°C, recombinant mutant F3S/S232G | Sulfurisphaera tokodaii | |
0.25 | - |
NAD+ | pH 7.5, 70°C, recombinant mutant R284G | Sulfurisphaera tokodaii | |
0.37 | - |
NAD+ | pH 7.5, 70°C, recombinant mutant L288P | Sulfurisphaera tokodaii | |
0.39 | - |
NAD+ | pH 7.5, 70°C, recombinant wild-type enzyme and mutant F3S | Sulfurisphaera tokodaii | |
0.4 | - |
NAD+ | pH 7.5, 70°C, recombinant mutant F3S/L288P | Sulfurisphaera tokodaii |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | - |
Sulfurisphaera tokodaii |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
(2R,3S)-3-isopropylmalate + NAD+ | Sulfurisphaera tokodaii | the enzyme is enzyme involved in leucine biosynthesis | ? | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Sulfurisphaera tokodaii | P50455 | gene leuB | - |
Purification (Comment) | Organism |
---|---|
recombinant wild-type and mutant enzymes from Escherichia coli strain MA153 by anion exchange and hydrophobic interaction chromatography | Sulfurisphaera tokodaii |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
(2R,3S)-3-isopropylmalate + NAD+ | - |
Sulfurisphaera tokodaii | ? | - |
? | |
(2R,3S)-3-isopropylmalate + NAD+ | the enzyme is enzyme involved in leucine biosynthesis | Sulfurisphaera tokodaii | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
tetramer | - |
Sulfurisphaera tokodaii |
Synonyms | Comment | Organism |
---|---|---|
IPMDH | - |
Sulfurisphaera tokodaii |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
95 | - |
- |
Sulfurisphaera tokodaii |
Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | 95 | 2% of maximal activity at 25°C, maximal activity at 95°C | Sulfurisphaera tokodaii |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
3.8 | - |
3-isopropylmalate | pH 7.5, 70°C, recombinant mutant F3S/S232G | Sulfurisphaera tokodaii | |
6 | - |
3-isopropylmalate | pH 7.5, 70°C, recombinant mutant R284G | Sulfurisphaera tokodaii | |
6.8 | - |
3-isopropylmalate | pH 7.5, 70°C, recombinant mutant F3S | Sulfurisphaera tokodaii | |
6.9 | - |
3-isopropylmalate | pH 7.5, 70°C, recombinant wild-type enzyme | Sulfurisphaera tokodaii | |
7.5 | - |
3-isopropylmalate | pH 7.5, 70°C, recombinant mutant L288P | Sulfurisphaera tokodaii | |
7.8 | - |
3-isopropylmalate | pH 7.5, 70°C, recombinant mutant F3S/L288P | Sulfurisphaera tokodaii |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
assay at | Sulfurisphaera tokodaii |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NAD+ | - |
Sulfurisphaera tokodaii |