Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli Rosetta 2(DE3)pLysS cells | Lysinibacillus sphaericus |
General Stability | Organism |
---|---|
the specific activity is increased nearly two times by Ni-NTA agarose gel purification | Lysinibacillus sphaericus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.38 | - |
3-methyl-2-oxopentanoate | in 100 mM glycine-KCl-KOH buffer (pH 10), at 25°C | Lysinibacillus sphaericus | |
0.78 | - |
L-Ile | in 100 mM glycine-KCl-KOH buffer (pH 10), at 25°C | Lysinibacillus sphaericus | |
0.98 | - |
L-norvaline | in 100 mM glycine-KCl-KOH buffer (pH 10), at 25°C | Lysinibacillus sphaericus | |
1.08 | - |
2-oxopentanoate | in 100 mM glycine-KCl-KOH buffer (pH 10), at 25°C | Lysinibacillus sphaericus | |
1.12 | - |
L-Leu | in 100 mM glycine-KCl-KOH buffer (pH 10), at 25°C | Lysinibacillus sphaericus | |
1.27 | - |
L-norleucine | in 100 mM glycine-KCl-KOH buffer (pH 10), at 25°C | Lysinibacillus sphaericus | |
1.55 | - |
4-methyl-2-oxopentanoate | in 100 mM glycine-KCl-KOH buffer (pH 10), at 25°C | Lysinibacillus sphaericus |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Lysinibacillus sphaericus | - |
strain ATCC4525 | - |
Lysinibacillus sphaericus ATCC 4525 | - |
strain ATCC4525 | - |
Purification (Comment) | Organism |
---|---|
Ni-NTA agarose gel chromatography | Lysinibacillus sphaericus |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
6.27 | - |
enzyme from cell-free extract, using L-Leu as substrate | Lysinibacillus sphaericus |
12.19 | - |
enzyme after Ni-NTA agaose gel purification, using L-Leu as substrate | Lysinibacillus sphaericus |
Storage Stability | Organism |
---|---|
4°C, purified enzyme without any stabilizer, 1 month, barely changed activity | Lysinibacillus sphaericus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
2-oxobutanoate + NH3 + NADH | 11% activity 2-oxobutanoate compared to 4-methyl-2-oxopentanoate | Lysinibacillus sphaericus | L-2-aminobutanoate + H2O + NAD+ | - |
r | |
2-oxohexanoate + NH3 + NADH | - |
Lysinibacillus sphaericus | L-norleucine + H2O + NAD+ | - |
r | |
2-oxopentanoate + NH3 + NADH | 25% activity with 2-oxopentanoate compared to 4-methyl-2-oxopentanoate | Lysinibacillus sphaericus | L-norvaline + H2O + NAD+ | - |
r | |
3-methyl-2-oxopentanoate + NH3 + NADH | 64% activity with 3-methyl-2-oxopentanoate compared to 4-methyl-2-oxopentanoate | Lysinibacillus sphaericus | L-Ile + H2O + NAD+ | - |
r | |
4-methyl-2-oxopentanoate + NH3 + NADH | 100% activity with 4-methyl-2-oxopentanoate | Lysinibacillus sphaericus | L-Leu + H2O + NAD+ | - |
r | |
4-methylthio-2-oxobutyrate + NH3 + NADH | 19% activity with 4-methylthio-2-oxobutyrate compared to 4-methyl-2-oxopentanoate | Lysinibacillus sphaericus | L-Met + H2O + NAD+ | - |
r | |
L-2-aminobutanoate + H2O + NAD+ | 5.7% activity wit L-2-aminobutanoate compared to L-Leu | Lysinibacillus sphaericus | 2-oxobutanoate + NH3 + NADH + H+ | - |
r | |
L-2-aminobutanoate + H2O + NAD+ | 5.7% activity wit L-2-aminobutanoate compared to L-Leu | Lysinibacillus sphaericus ATCC 4525 | 2-oxobutanoate + NH3 + NADH + H+ | - |
r | |
L-Ala + H2O + NAD+ | 3.4% activity with L-Ala compared to L-Leu | Lysinibacillus sphaericus | 2-oxopropanoate + NH3 + NADH | - |
r | |
L-Ile + H2O + NAD+ | 72.4% activity with L-Ile compared to L-Leu | Lysinibacillus sphaericus | 3-methyl-2-oxopentanoate + NH3 + NADH | - |
r | |
L-Ile + H2O + NAD+ | 72.4% activity with L-Ile compared to L-Leu | Lysinibacillus sphaericus ATCC 4525 | 3-methyl-2-oxopentanoate + NH3 + NADH | - |
r | |
L-Leu + H2O + NAD+ | 100% activity with L-Leu | Lysinibacillus sphaericus | 4-methyl-2-oxopentanoate + NH3 + NADH | - |
r | |
L-Leu + H2O + NAD+ | 100% activity with L-Leu | Lysinibacillus sphaericus ATCC 4525 | 4-methyl-2-oxopentanoate + NH3 + NADH | - |
r | |
L-Met + H2O + NAD+ | 1.2% activity with L-Met compared to L-Leu | Lysinibacillus sphaericus | 4-methylthio-2-oxobutyrate + NH3 + NADH + H+ | - |
r | |
L-norleucine + H2O + NAD+ | - |
Lysinibacillus sphaericus | 2-oxohexanoate + NH3 + NADH | - |
r | |
L-norvaline + H2O + NAD+ | 36.8% activity with L-norvaline compared to L-Leu | Lysinibacillus sphaericus | 2-oxopentanoate + NH3 + NADH | - |
r | |
L-norvaline + H2O + NAD+ | 36.8% activity with L-norvaline compared to L-Leu | Lysinibacillus sphaericus ATCC 4525 | 2-oxopentanoate + NH3 + NADH | - |
r | |
additional information | no activity with L-Cys, L-Phe, 2-oxo-3-phenylpropionic acid, 3-(2'-nitrophenyl)-2-oxopropionic acid, and 3-(3'-indolyl)-2-oxopropionic acid | Lysinibacillus sphaericus | ? | - |
? | |
additional information | no activity with L-Cys, L-Phe, 2-oxo-3-phenylpropionic acid, 3-(2'-nitrophenyl)-2-oxopropionic acid, and 3-(3'-indolyl)-2-oxopropionic acid | Lysinibacillus sphaericus ATCC 4525 | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
LeuDH | - |
Lysinibacillus sphaericus |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.18 | - |
L-norleucine | in 100 mM glycine-KCl-KOH buffer (pH 10), at 25°C | Lysinibacillus sphaericus | |
1.53 | - |
L-norvaline | in 100 mM glycine-KCl-KOH buffer (pH 10), at 25°C | Lysinibacillus sphaericus | |
3.31 | - |
L-Ile | in 100 mM glycine-KCl-KOH buffer (pH 10), at 25°C | Lysinibacillus sphaericus | |
10.83 | - |
L-Leu | in 100 mM glycine-KCl-KOH buffer (pH 10), at 25°C | Lysinibacillus sphaericus | |
27.63 | - |
3-methyl-2-oxopentanoate | in 100 mM glycine-KCl-KOH buffer (pH 10), at 25°C | Lysinibacillus sphaericus | |
47.69 | - |
2-oxopentanoate | in 100 mM glycine-KCl-KOH buffer (pH 10), at 25°C | Lysinibacillus sphaericus | |
99.38 | - |
4-methyl-2-oxopentanoate | in 100 mM glycine-KCl-KOH buffer (pH 10), at 25°C | Lysinibacillus sphaericus |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NAD+ | - |
Lysinibacillus sphaericus |