Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli BL21(DE3) cells | Alcaligenes faecalis |
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 | 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 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 | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
acetoacetate + NADH + H+ | Alcaligenes faecalis | - |
(R)-3-hydroxybutanoate + NAD+ | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Alcaligenes faecalis | D0VWQ0 | - |
- |
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 | Comment | Organism |
---|---|---|
3HBDH | - |
Alcaligenes faecalis |
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 | Comment | Organism | Structure |
---|---|---|---|
NADH | - |
Alcaligenes faecalis |
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 |