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Literature summary for 1.1.1.86 extracted from

  • Dumas, R.; Biou, V.; Douce, R.
    Purification and characterization of a fusion protein of plant acetohydroxy acid synthase and acetohydroxy acid isomeroreductase (1997), FEBS Lett., 408, 156-160.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
the nucleotide sequence coding for the Arabidopsis thaliana acetohydroxy acid synthase is genetically fused in frame with the nucleotide sequence coding for the Spinacia oleracea acetohydroxy acid isomeroreductase and expressed in Escherichia coli Spinacia oleracea

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.007
-
NADPH pH 8.2, 30°C, chimeric enzyme Arabidopsis thaliana acetohydroxy acid synthase is genetically fused in frame with the nucleotide sequence coding for the Spinacia oleracea acetohydroxy acid isomeroreductase and expressed in E. coli Spinacia oleracea
0.01
-
2-aceto-2-hydroxybutanoate pH 8.2, 30°C, chimeric enzyme Aabidopsis thaliana acetohydroxy acid synthase genetically fused in frame with the nucleotide sequence coding for the Spinacia oleracea acetohydroxy acid isomeroreductase and expressed in E. coli Spinacia oleracea

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+ required Spinacia oleracea
Mg2+ Km: 0.5 mM Spinacia oleracea

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
120000
-
2 * 120000, Arabidopsis thaliana acetohydroxy acid synthase genetically fused in frame with the nucleotide sequence coding for the Spinacia oleracea acetohydroxy acid isomeroreductase and expressed in Escherichia coli, SDS-PAGE Spinacia oleracea
240000
-
Arabidopsis thaliana acetohydroxy acid synthase genetically fused in frame with the nucleotide sequence coding for the Spinacia oleracea acetohydroxy acid isomeroreductase and expressed in E. coli, gel filtration Spinacia oleracea

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2-acetolactate + NADPH Spinacia oleracea enzyme of branched chain amino acid synthesis 2,3-dihydroxy-3-methylbutanoate + NADP+
-
r

Organism

Organism UniProt Comment Textmining
Spinacia oleracea
-
overexpressed in Escherichia coli
-

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.9
-
Arabidopsis thaliana acetohydroxy acid synthase is genetically fused in frame with the nucleotide sequence coding for the Spinacia oleracea acetohydroxy acid isomeroreductase and expressed in E. coli Spinacia oleracea

Storage Stability

Storage Stability Organism
-80°, chimeric enzyme composed of Arabidopsis thaliana acetohydroxy acid synthase genetically fused in frame with the nucleotide sequence coding for the Spinacia oleracea acetohydroxy acid isomeroreductase and expressed in E. coli, several months, no loss of activity Spinacia oleracea

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2-acetolactate + NADPH enzyme of branched chain amino acid synthesis Spinacia oleracea 2,3-dihydroxy-3-methylbutanoate + NADP+
-
r
NADPH + 2-aceto-2-hydroxybutyrate
-
Spinacia oleracea NADP+ + ?
-
?
NADPH + 2-acetolactate
-
Spinacia oleracea NADP+ + 3-hydroxy-3-methyl-2-oxobutyrate
-
?

Subunits

Subunits Comment Organism
dimer 2 * 120000, Arabidopsis thaliana acetohydroxy acid synthase genetically fused in frame with the nucleotide sequence coding for the Spinacia oleracea acetohydroxy acid isomeroreductase and expressed in Escherichia coli, SDS-PAGE Spinacia oleracea

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8.2
-
isomeroreductase activity of the chimeric enzyme composed of Arabidopsis thaliana acetohydroxy acid synthase genetically fused in frame with the nucleotide sequence coding for the Spinacia oleracea acetohydroxy acid isomeroreductase and expressed in Escherichia coli Spinacia oleracea

Cofactor

Cofactor Comment Organism Structure
NADPH
-
Spinacia oleracea