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Literature summary 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)

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

Protein Variants

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

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.1.1.35 0.03
-
 acetoacetyl-CoA pH 5.0, recombinant His-tagged mutant S137C Rattus norvegicus
1.1.1.35 0.044
-
 acetoacetyl-CoA pH 5.0, recombinant His-tagged wild-type enzyme Rattus norvegicus
1.1.1.35 0.184
-
 acetoacetyl-CoA pH 5.0, recombinant His-tagged mutant S137A Rattus norvegicus
1.1.1.35 0.263
-
 acetoacetyl-CoA pH 5.0, recombinant His-tagged mutant S137T Rattus norvegicus

Localization

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

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.1.1.35 3-oxoacyl-CoA + NADH Rattus norvegicus key enzyme involved in fatty acid oxidation 3-hydroxyacyl-CoA + NAD+
-
 r

Organism

EC Number Organism UniProt Comment Textmining
1.1.1.35 Rattus norvegicus
-
-
-

Purification (Commentary)

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

Reaction

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

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.1.1.35 additional information
-
 the N-terminal His-tag does notinfluence enzyme activity Rattus norvegicus
1.1.1.35 452
-
 purified recombinant His-tagged wild-type enzyme Rattus norvegicus

Storage Stability

EC Number Storage Stability Organism
1.1.1.35 -80°C, purified recombinant His-tagged wild-type and mutant enzymes, stable for at least 3 months Rattus norvegicus
1.1.1.35 4°C, purified recombinant His-tagged wild-type enzyme, 1 week, no significant loss of activity Rattus norvegicus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.1.1.35 3-oxoacyl-CoA + NADH key enzyme involved in fatty acid oxidation Rattus norvegicus 3-hydroxyacyl-CoA + NAD+
-
 r
1.1.1.35 acetoacetyl-CoA + NADH + H+
-
 Rattus norvegicus 3-hydroxybutyryl-CoA + NAD+
-
 r

Synonyms

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

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.1.1.35 0.019
-
 acetoacetyl-CoA pH 5.0, recombinant His-tagged mutant S137C Rattus norvegicus
1.1.1.35 0.026
-
 acetoacetyl-CoA pH 5.0, recombinant His-tagged mutant S137T Rattus norvegicus
1.1.1.35 0.12
-
 acetoacetyl-CoA pH 5.0, recombinant His-tagged mutant S137A Rattus norvegicus
1.1.1.35 569
-
 acetoacetyl-CoA pH 5.0, recombinant His-tagged wild-type enzyme Rattus norvegicus

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.1.1.35 5
-
 phosphate buffer Rattus norvegicus

Cofactor

EC Number Cofactor Comment Organism Structure
1.1.1.35 NAD+
-
 Rattus norvegicus
1.1.1.35 NADH
-
 Rattus norvegicus