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Literature summary extracted from

  • Li, H.; Zhu, D.; Hyatt, B.A.; Malik, F.M.; Biehl, E.R.; Hua, L.
    Cloning, protein sequence clarification, and substrate specificity of a leucine dehydrogenase from Bacillus sphaericus ATCC4525 (2008), Appl. Biochem. Biotechnol., 158, 343-351.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.4.1.9 expressed in Escherichia coli Rosetta 2(DE3)pLysS cells Lysinibacillus sphaericus

General Stability

EC Number General Stability Organism
1.4.1.9 the specific activity is increased nearly two times by Ni-NTA agarose gel purification Lysinibacillus sphaericus

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.4.1.9 0.38
-
 3-methyl-2-oxopentanoate in 100 mM glycine-KCl-KOH buffer (pH 10), at 25°C Lysinibacillus sphaericus
1.4.1.9 0.78
-
 L-Ile in 100 mM glycine-KCl-KOH buffer (pH 10), at 25°C Lysinibacillus sphaericus
1.4.1.9 0.98
-
 L-norvaline in 100 mM glycine-KCl-KOH buffer (pH 10), at 25°C Lysinibacillus sphaericus
1.4.1.9 1.08
-
 2-oxopentanoate in 100 mM glycine-KCl-KOH buffer (pH 10), at 25°C Lysinibacillus sphaericus
1.4.1.9 1.12
-
 L-Leu in 100 mM glycine-KCl-KOH buffer (pH 10), at 25°C Lysinibacillus sphaericus
1.4.1.9 1.27
-
 L-norleucine in 100 mM glycine-KCl-KOH buffer (pH 10), at 25°C Lysinibacillus sphaericus
1.4.1.9 1.55
-
 4-methyl-2-oxopentanoate in 100 mM glycine-KCl-KOH buffer (pH 10), at 25°C Lysinibacillus sphaericus

Organism

EC Number Organism UniProt Comment Textmining
1.4.1.9 Lysinibacillus sphaericus
-
 strain ATCC4525
-
1.4.1.9 Lysinibacillus sphaericus ATCC 4525
-
 strain ATCC4525
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.4.1.9 Ni-NTA agarose gel chromatography Lysinibacillus sphaericus

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.4.1.9 6.27
-
 enzyme from cell-free extract, using L-Leu as substrate Lysinibacillus sphaericus
1.4.1.9 12.19
-
 enzyme after Ni-NTA agaose gel purification, using L-Leu as substrate Lysinibacillus sphaericus

Storage Stability

EC Number Storage Stability Organism
1.4.1.9 4°C, purified enzyme without any stabilizer, 1 month, barely changed activity Lysinibacillus sphaericus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.4.1.9 2-oxobutanoate + NH3 + NADH 11% activity 2-oxobutanoate compared to 4-methyl-2-oxopentanoate Lysinibacillus sphaericus L-2-aminobutanoate + H2O + NAD+
-
 r
1.4.1.9 2-oxohexanoate + NH3 + NADH
-
 Lysinibacillus sphaericus L-norleucine + H2O + NAD+
-
 r
1.4.1.9 2-oxopentanoate + NH3 + NADH 25% activity with 2-oxopentanoate compared to 4-methyl-2-oxopentanoate Lysinibacillus sphaericus L-norvaline + H2O + NAD+
-
 r
1.4.1.9 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
1.4.1.9 4-methyl-2-oxopentanoate + NH3 + NADH 100% activity with 4-methyl-2-oxopentanoate Lysinibacillus sphaericus L-Leu + H2O + NAD+
-
 r
1.4.1.9 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
1.4.1.9 L-2-aminobutanoate + H2O + NAD+ 5.7% activity wit L-2-aminobutanoate compared to L-Leu Lysinibacillus sphaericus 2-oxobutanoate + NH3 + NADH + H+
-
 r
1.4.1.9 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
1.4.1.9 L-Ala + H2O + NAD+ 3.4% activity with L-Ala compared to L-Leu Lysinibacillus sphaericus 2-oxopropanoate + NH3 + NADH
-
 r
1.4.1.9 L-Ile + H2O + NAD+ 72.4% activity with L-Ile compared to L-Leu Lysinibacillus sphaericus 3-methyl-2-oxopentanoate + NH3 + NADH
-
 r
1.4.1.9 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
1.4.1.9 L-Leu + H2O + NAD+ 100% activity with L-Leu Lysinibacillus sphaericus 4-methyl-2-oxopentanoate + NH3 + NADH
-
 r
1.4.1.9 L-Leu + H2O + NAD+ 100% activity with L-Leu Lysinibacillus sphaericus ATCC 4525 4-methyl-2-oxopentanoate + NH3 + NADH
-
 r
1.4.1.9 L-Met + H2O + NAD+ 1.2% activity with L-Met compared to L-Leu Lysinibacillus sphaericus 4-methylthio-2-oxobutyrate + NH3 + NADH + H+
-
 r
1.4.1.9 L-norleucine + H2O + NAD+
-
 Lysinibacillus sphaericus 2-oxohexanoate + NH3 + NADH
-
 r
1.4.1.9 L-norvaline + H2O + NAD+ 36.8% activity with L-norvaline compared to L-Leu Lysinibacillus sphaericus 2-oxopentanoate + NH3 + NADH
-
 r
1.4.1.9 L-norvaline + H2O + NAD+ 36.8% activity with L-norvaline compared to L-Leu Lysinibacillus sphaericus ATCC 4525 2-oxopentanoate + NH3 + NADH
-
 r
1.4.1.9 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 ?
-
 ?
1.4.1.9 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

EC Number Synonyms Comment Organism
1.4.1.9 LeuDH
-
 Lysinibacillus sphaericus

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.4.1.9 0.18
-
 L-norleucine in 100 mM glycine-KCl-KOH buffer (pH 10), at 25°C Lysinibacillus sphaericus
1.4.1.9 1.53
-
 L-norvaline in 100 mM glycine-KCl-KOH buffer (pH 10), at 25°C Lysinibacillus sphaericus
1.4.1.9 3.31
-
 L-Ile in 100 mM glycine-KCl-KOH buffer (pH 10), at 25°C Lysinibacillus sphaericus
1.4.1.9 10.83
-
 L-Leu in 100 mM glycine-KCl-KOH buffer (pH 10), at 25°C Lysinibacillus sphaericus
1.4.1.9 27.63
-
 3-methyl-2-oxopentanoate in 100 mM glycine-KCl-KOH buffer (pH 10), at 25°C Lysinibacillus sphaericus
1.4.1.9 47.69
-
 2-oxopentanoate in 100 mM glycine-KCl-KOH buffer (pH 10), at 25°C Lysinibacillus sphaericus
1.4.1.9 99.38
-
 4-methyl-2-oxopentanoate in 100 mM glycine-KCl-KOH buffer (pH 10), at 25°C Lysinibacillus sphaericus

Cofactor

EC Number Cofactor Comment Organism Structure
1.4.1.9 NAD+
-
 Lysinibacillus sphaericus