Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary for 1.1.1.103 extracted from

  • Ueatrongchit, T.; Asano, Y.
    Highly selective L-threonine 3-dehydrogenase from Cupriavidus necator and its use in determination of L-threonine (2011), Anal. Biochem., 410, 44-56.
    View publication on PubMed

Activating Compound

Activating Compound Comment Organism Structure
2-mercaptoethanol 14% activation at 10 mM Cupriavidus necator
DTT 25% activation at 10 mM Cupriavidus necator

Cloned(Commentary)

Cloned (Comment) Organism
DNA and amino acid seuence determination and analysis, sequence comparisons, phylogenetic tree, expression of His-tagged enzyme in Escherichia coli strains JM109 and BL21(DE3) Cupriavidus necator

Inhibitors

Inhibitors Comment Organism Structure
4-chloromercuribenzonic acid 94% inhibition at 10 mM Cupriavidus necator
calcium pantothenate slight inhibition Cupriavidus necator
FeCl2 32% inhibition at 1 mM after 60 min Cupriavidus necator
FeCl3 73% inhibition at 1 mM after 60 min Cupriavidus necator
HgCl2 77% inhibition at 1 mM after 60 min Cupriavidus necator
iodoacetamide 99% inhibition at 10 mM Cupriavidus necator
iodoacetic acid 52% inhibition at 10 mM Cupriavidus necator
K3[Fe(CN)6] 25% inhibition at 10 mM Cupriavidus necator
additional information poor inhibition by K[Fe(CN)6], no inhibition by ethylene diamine tetraacetic acid and ethylene glycol tetraacetic acid, and by trypsin inhibitor T-9378 Cupriavidus necator
N-ethylmaleimide complete inhibition Cupriavidus necator
NaN3 88% inhibition at 10 mM Cupriavidus necator
phenazinemethosulfate complete inhibition Cupriavidus necator
phenylmethanesulfonyl fluoride 63% inhibition at 10 mM Cupriavidus necator
SnCl2 16% inhibition at 1 mM after 60 min Cupriavidus necator

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.1
-
NAD+ pH 10.0, 30°C, native enzyme Cupriavidus necator
11.6
-
L-threonine pH 10.0, 30°C, native enzyme Cupriavidus necator

Metals/Ions

Metals/Ions Comment Organism Structure
AlCl3 18% activation at 1 mM Cupriavidus necator
BaCl2 13% activation at 1 mM Cupriavidus necator
CoCl2 15% activation at 1 mM Cupriavidus necator
CrCl3 10% activation at 1 mM Cupriavidus necator
CsCl 17% activation at 1 mM Cupriavidus necator
CuSO4 15% activation at 1 mM Cupriavidus necator
MgSO4 12% activation at 1 mM Cupriavidus necator
MnSO4 12% activation at 1 mM Cupriavidus necator
additional information no effect at 1 mM by AgNO3, MgCl2, and 10 mM EDTA and ethylene glycol tetraacetic acid Cupriavidus necator
Na2MoO4 13% activation at 1 mM Cupriavidus necator
NiCl2 19% activation at 1 mM Cupriavidus necator
PbCl2 18% activation at 1 mM Cupriavidus necator
ZnSO4 9% activation at 1 mM Cupriavidus necator

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
34628
-
2 x 37200, native enzyme, SDS-PAGE, x * 34628, sequence calculation Cupriavidus necator
79400
-
native enzyme, gel filtration Cupriavidus necator

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
L-threonine + NAD+ Cupriavidus necator
-
(2S)-2-amino-3-oxobutanoate + NADH + H+
-
?
L-threonine + NAD+ Cupriavidus necator NBRC 102504
-
(2S)-2-amino-3-oxobutanoate + NADH + H+
-
?

Organism

Organism UniProt Comment Textmining
Cupriavidus necator E5RQ20
-
-
Cupriavidus necator NBRC 102504 E5RQ20
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant His-tagged enzyme 4.2fold from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, native enzyme 75.4fold by anion exchange and hydrophobic interaction chromatography, dialysis, hydroxyapatite chromatography, followed by gel filtration Cupriavidus necator

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
42.2
-
purified native enzyme, pH 10.0, 30°C Cupriavidus necator
65
-
purified recombinant enzyme, pH 10.0, 30°C Cupriavidus necator

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
DL-2-amino-3-hydroxyvalerate + NAD+
-
Cupriavidus necator DL-2-amino-3-oxopentanoate + NADH + H+
-
?
DL-2-amino-3-hydroxyvalerate + NAD+
-
Cupriavidus necator NBRC 102504 DL-2-amino-3-oxopentanoate + NADH + H+
-
?
L-threonine + NAD+
-
Cupriavidus necator (2S)-2-amino-3-oxobutanoate + NADH + H+
-
?
L-threonine + NAD+
-
Cupriavidus necator NBRC 102504 (2S)-2-amino-3-oxobutanoate + NADH + H+
-
?
additional information L-threonine and DL-2-amino-3-hydroxyvalerate are the only substrates for ThrDH among other L-amino acids, alcohols, and amino alcohols, substrate specificity, overview Cupriavidus necator ?
-
?
additional information L-threonine and DL-2-amino-3-hydroxyvalerate are the only substrates for ThrDH among other L-amino acids, alcohols, and amino alcohols, substrate specificity, overview Cupriavidus necator NBRC 102504 ?
-
?

Subunits

Subunits Comment Organism
dimer 2 x 37200, native enzyme, SDS-PAGE, x * 34628, sequence calculation Cupriavidus necator

Synonyms

Synonyms Comment Organism
ThrDH
-
Cupriavidus necator

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
75
-
-
Cupriavidus necator

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
25 80 activity range, profile overview Cupriavidus necator

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
25 75 quite stable at, rapid loss of activity above 80°C Cupriavidus necator

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
38
-
NAD+ pH 10.0, 30°C, native enzyme Cupriavidus necator
43.8
-
L-threonine pH 10.0, 30°C, native enzyme Cupriavidus necator

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
10
-
glycine-KOH buffer Cupriavidus necator

pH Range

pH Minimum pH Maximum Comment Organism
6 12 activity range, pH-dependent activity varies in different buffer systems, profile overview Cupriavidus necator

pH Stability

pH Stability pH Stability Maximum Comment Organism
4 11 the purified enzyme is very stable from pH 6.0 to pH 11.0 in the glycine-KOH system, the enzyme is unstable in sodium acetate buffer, pH 4.0-5.0, and Na2CO3-NaHCO3 buffer, pH 10.0-11.0 Cupriavidus necator

Cofactor

Cofactor Comment Organism Structure
NAD+ dependent on Cupriavidus necator

General Information

General Information Comment Organism
evolution the enzyme belongs to the extended short-chain alcohol dehydrogenase superfamily Cupriavidus necator
additional information the enzyme possesses a glycine-rich NAD+-binding domain at the N terminus and conserved catalytic triad of YxxxK residues Cupriavidus necator