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Literature summary extracted from

  • Yeon, Y.; Park, H.; Park, H.; Yoo, Y.
    Effect of His-tag location on the catalytic activity of 3-hydroxybutyrate dehydrogenase (2014), Biotechnol. Bioprocess Eng., 19, 798-802.
No PubMed abstract available

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.1.1.30 expressed in Escherichia coli BL21(DE3) cells Alcaligenes faecalis

Protein Variants

EC Number Protein Variants Comment Organism
1.1.1.30 H144L the mutant shows activity with levulinic acid Alcaligenes faecalis
1.1.1.30 H144L/W187F the mutant shows activity with levulinic acid Alcaligenes faecalis
1.1.1.30 W187F the mutant shows activity with levulinic acid Alcaligenes faecalis

General Stability

EC Number General Stability Organism
1.1.1.30 counterparts Alcaligenes faecalis
1.1.1.30 the enzyme with an N-terminal His-tag has approximately 1200fold higher catalytic efficiency than its C-terminal counterpart and exhibits higher catalytic efficiency for levulinic acid than the C-terminal Alcaligenes faecalis

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.1.1.30 0.17
-
 acetoacetate recombinant enzyme with N-terminal His-tag, at pH 6.5 and 30°C Alcaligenes faecalis
1.1.1.30 0.63
-
 levulinic acid recombinant mutant enzyme W187F with N-terminal His-tag, at pH 6.5 and 30°C Alcaligenes faecalis
1.1.1.30 0.64
-
 levulinic acid recombinant mutant enzyme H144L/W187F with N-terminal His-tag, at pH 6.5 and 30°C Alcaligenes faecalis
1.1.1.30 0.78
-
 levulinic acid recombinant wild type enzyme with N-terminal His-tag, at pH 6.5 and 30°C Alcaligenes faecalis
1.1.1.30 0.91
-
 levulinic acid recombinant mutant enzyme H144L with N-terminal His-tag, at pH 6.5 and 30°C Alcaligenes faecalis
1.1.1.30 8.25
-
 levulinic acid recombinant mutant enzyme H144L/W187F with C-terminal His-tag, at pH 6.5 and 30°C Alcaligenes faecalis
1.1.1.30 10.47
-
 acetoacetate recombinant enzyme with C-terminal His-tag, at pH 6.5 and 30°C Alcaligenes faecalis
1.1.1.30 16.98
-
 levulinic acid recombinant mutant enzyme H144L with C-terminal His-tag, at pH 6.5 and 30°C Alcaligenes faecalis

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.1.1.30 acetoacetate + NADH + H+ Alcaligenes faecalis
-
 (R)-3-hydroxybutanoate + NAD+
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
1.1.1.30 Alcaligenes faecalis D0VWQ0
-
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.1.1.30 acetoacetate + NADH + H+
-
 Alcaligenes faecalis (R)-3-hydroxybutanoate + NAD+
-
 ?
1.1.1.30 levulinic acid + NADH + H+ only the recombinant wild type enzyme with N-terminal His-tag shows activity with levulinic acid Alcaligenes faecalis 4-hydroxyvaleric acid + NAD+
-
 ?

Synonyms

EC Number Synonyms Comment Organism
1.1.1.30 3HBDH
-
 Alcaligenes faecalis

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.1.1.30 0.006
-
 levulinic acid recombinant wild type enzyme with N-terminal His-tag, at pH 6.5 and 30°C Alcaligenes faecalis
1.1.1.30 0.019
-
 levulinic acid recombinant mutant enzyme W187F with N-terminal His-tag, at pH 6.5 and 30°C Alcaligenes faecalis
1.1.1.30 0.028
-
 levulinic acid recombinant mutant enzyme H144L with C-terminal His-tag, at pH 6.5 and 30°C Alcaligenes faecalis
1.1.1.30 0.06
-
 levulinic acid recombinant mutant enzyme H144L with N-terminal His-tag, at pH 6.5 and 30°C Alcaligenes faecalis
1.1.1.30 0.079
-
 levulinic acid recombinant mutant enzyme H144L/W187F with C-terminal His-tag, at pH 6.5 and 30°C Alcaligenes faecalis
1.1.1.30 0.164
-
 levulinic acid recombinant mutant enzyme H144L/W187F with N-terminal His-tag, at pH 6.5 and 30°C Alcaligenes faecalis
1.1.1.30 4.9
-
 acetoacetate recombinant enzyme with C-terminal His-tag, at pH 6.5 and 30°C Alcaligenes faecalis
1.1.1.30 97.16
-
 acetoacetate recombinant enzyme with N-terminal His-tag, at pH 6.5 and 30°C Alcaligenes faecalis

Cofactor

EC Number Cofactor Comment Organism Structure
1.1.1.30 NADH
-
 Alcaligenes faecalis

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1.1.1.30 0.47
-
 acetoacetate recombinant enzyme with C-terminal His-tag, at pH 6.5 and 30°C Alcaligenes faecalis
1.1.1.30 1.62
-
 levulinic acid recombinant mutant enzyme H144L with C-terminal His-tag, at pH 6.5 and 30°C Alcaligenes faecalis
1.1.1.30 7.68
-
 levulinic acid recombinant wild type enzyme with N-terminal His-tag, at pH 6.5 and 30°C Alcaligenes faecalis
1.1.1.30 9.63
-
 levulinic acid recombinant mutant enzyme H144L/W187F with C-terminal His-tag, at pH 6.5 and 30°C Alcaligenes faecalis
1.1.1.30 29.69
-
 levulinic acid recombinant mutant enzyme W187F with N-terminal His-tag, at pH 6.5 and 30°C Alcaligenes faecalis
1.1.1.30 65.94
-
 levulinic acid recombinant mutant enzyme H144L with N-terminal His-tag, at pH 6.5 and 30°C Alcaligenes faecalis
1.1.1.30 256.5
-
 levulinic acid recombinant mutant enzyme H144L/W187F with N-terminal His-tag, at pH 6.5 and 30°C Alcaligenes faecalis
1.1.1.30 560.1
-
 acetoacetate recombinant enzyme with N-terminal His-tag, at pH 6.5 and 30°C Alcaligenes faecalis