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Literature summary for 1.1.1.30 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)

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

Protein Variants

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

General Stability

General Stability Organism
counterparts Alcaligenes faecalis
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]

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

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

Organism

Organism UniProt Comment Textmining
Alcaligenes faecalis D0VWQ0
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
acetoacetate + NADH + H+
-
Alcaligenes faecalis (R)-3-hydroxybutanoate + NAD+
-
?
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

Synonyms Comment Organism
3HBDH
-
Alcaligenes faecalis

Turnover Number [1/s]

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

Cofactor

Cofactor Comment Organism Structure
NADH
-
Alcaligenes faecalis

kcat/KM [mM/s]

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