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Literature summary for 1.1.1.35 extracted from

  • Liu, X.; Chu, X.; Yu, W.; Li, P.; Li, D.
    Expression and purification of His-tagged rat mitochondrial short-chain 3-hydroxyacyl-CoA dehydrogenase wild-type and Ser137 mutant proteins (2004), Protein Expr. Purif., 37, 344-351.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
overexpression of wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) as soluble N-terminally His-tagged enzymes Rattus norvegicus

Protein Variants

Protein Variants Comment Organism
S137A site-directed mutagenesis, highly reduced activity compared to the wild-type enzyme Rattus norvegicus
S137C site-directed mutagenesis, highly reduced activity compared to the wild-type enzyme Rattus norvegicus
S137T site-directed mutagenesis, highly reduced activity compared to the wild-type enzyme Rattus norvegicus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.03
-
acetoacetyl-CoA pH 5.0, recombinant His-tagged mutant S137C Rattus norvegicus
0.044
-
acetoacetyl-CoA pH 5.0, recombinant His-tagged wild-type enzyme Rattus norvegicus
0.184
-
acetoacetyl-CoA pH 5.0, recombinant His-tagged mutant S137A Rattus norvegicus
0.263
-
acetoacetyl-CoA pH 5.0, recombinant His-tagged mutant S137T Rattus norvegicus

Localization

Localization Comment Organism GeneOntology No. Textmining
mitochondrion
-
Rattus norvegicus 5739
-

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
3-oxoacyl-CoA + NADH Rattus norvegicus key enzyme involved in fatty acid oxidation 3-hydroxyacyl-CoA + NAD+
-
r

Organism

Organism UniProt Comment Textmining
Rattus norvegicus
-
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli by nickel affinity chromatography Rattus norvegicus

Reaction

Reaction Comment Organism Reaction ID
(S)-3-hydroxyacyl-CoA + NAD+ = 3-oxoacyl-CoA + NADH + H+ highly conserved residue Ser137 is essential for activity Rattus norvegicus

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
additional information
-
the N-terminal His-tag does notinfluence enzyme activity Rattus norvegicus
452
-
purified recombinant His-tagged wild-type enzyme Rattus norvegicus

Storage Stability

Storage Stability Organism
-80°C, purified recombinant His-tagged wild-type and mutant enzymes, stable for at least 3 months Rattus norvegicus
4°C, purified recombinant His-tagged wild-type enzyme, 1 week, no significant loss of activity Rattus norvegicus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3-oxoacyl-CoA + NADH key enzyme involved in fatty acid oxidation Rattus norvegicus 3-hydroxyacyl-CoA + NAD+
-
r
acetoacetyl-CoA + NADH + H+
-
Rattus norvegicus 3-hydroxybutyryl-CoA + NAD+
-
r

Synonyms

Synonyms Comment Organism
short-chain 3-hydroxyacyl-CoA dehydrogenase
-
Rattus norvegicus

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.019
-
acetoacetyl-CoA pH 5.0, recombinant His-tagged mutant S137C Rattus norvegicus
0.026
-
acetoacetyl-CoA pH 5.0, recombinant His-tagged mutant S137T Rattus norvegicus
0.12
-
acetoacetyl-CoA pH 5.0, recombinant His-tagged mutant S137A Rattus norvegicus
569
-
acetoacetyl-CoA pH 5.0, recombinant His-tagged wild-type enzyme Rattus norvegicus

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
5
-
phosphate buffer Rattus norvegicus

Cofactor

Cofactor Comment Organism Structure
NAD+
-
Rattus norvegicus
NADH
-
Rattus norvegicus