Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli BL21(DE3) cells | Alcaligenes faecalis |
Protein Variants | Comment | Organism |
---|---|---|
H144A | the mutant shows reduced catalytic efficiency with levulinic acid compared to the wild type enzyme | Alcaligenes faecalis |
H144F | the mutant shows increased catalytic efficiency with levulinic acid compared to the wild type enzyme | Alcaligenes faecalis |
H144I | the mutant shows increased catalytic efficiency with levulinic acid compared to the wild type enzyme | Alcaligenes faecalis |
H144L | the mutant shows increased catalytic efficiency with levulinic acid compared to the wild type enzyme | Alcaligenes faecalis |
H144L/W187A | the mutant shows reduced catalytic efficiency with levulinic acid compared to the wild type enzyme | Alcaligenes faecalis |
H144L/W187F | the mutant shows strongly increased catalytic efficiency with levulinic acid compared to the wild type enzyme | Alcaligenes faecalis |
H144L/W187I | the mutant shows reduced catalytic efficiency with levulinic acid compared to the wild type enzyme | Alcaligenes faecalis |
H144L/W187L | the mutant shows strongly increased catalytic efficiency with levulinic acid compared to the wild type enzyme | Alcaligenes faecalis |
H144L/W187V | the mutant shows reduced catalytic efficiency with levulinic acid compared to the wild type enzyme | Alcaligenes faecalis |
H144V | the mutant shows wild type catalytic efficiency with levulinic acid | Alcaligenes faecalis |
W187A | the mutant shows reduced catalytic efficiency with levulinic acid compared to the wild type enzyme | Alcaligenes faecalis |
W187F | the mutant shows increased catalytic efficiency with levulinic acid compared to the wild type enzyme | Alcaligenes faecalis |
W187I | the mutant shows reduced catalytic efficiency with levulinic acid compared to the wild type enzyme | Alcaligenes faecalis |
W187L | the mutant shows reduced catalytic efficiency with levulinic acid compared to the wild type enzyme | Alcaligenes faecalis |
W187V | the mutant shows reduced catalytic efficiency with levulinic acid compared to the wild type enzyme | Alcaligenes faecalis |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.63 | - |
levulinic acid | mutant enzyme H144L/W187L, at pH 6.5 and 30°C | Alcaligenes faecalis | |
0.63 | - |
levulinic acid | mutant enzyme W187F, at pH 6.5 and 30°C | Alcaligenes faecalis | |
0.64 | - |
levulinic acid | mutant enzyme H144L/W187F, at pH 6.5 and 30°C | Alcaligenes faecalis | |
0.78 | - |
levulinic acid | wild type enzyme, at pH 6.5 and 30°C | Alcaligenes faecalis | |
0.91 | - |
levulinic acid | mutant enzyme H144L, at pH 6.5 and 30°C | Alcaligenes faecalis | |
1.72 | - |
levulinic acid | mutant enzyme H144F, at pH 6.5 and 30°C | Alcaligenes faecalis | |
2.81 | - |
levulinic acid | mutant enzyme H144I, at pH 6.5 and 30°C | Alcaligenes faecalis | |
3.84 | - |
levulinic acid | mutant enzyme H144V, at pH 6.5 and 30°C | Alcaligenes faecalis | |
4 | 5.4 | levulinic acid | mutant enzyme W187I, at pH 6.5 and 30°C | Alcaligenes faecalis | |
9.87 | - |
levulinic acid | mutant enzyme H144A, at pH 6.5 and 30°C | Alcaligenes faecalis | |
13.03 | - |
levulinic acid | mutant enzyme H144L/W187A, at pH 6.5 and 30°C | Alcaligenes faecalis | |
13.49 | - |
levulinic acid | mutant enzyme W187L, at pH 6.5 and 30°C | Alcaligenes faecalis | |
25.57 | - |
levulinic acid | mutant enzyme H144L/W187V, at pH 6.5 and 30°C | Alcaligenes faecalis | |
25.76 | - |
levulinic acid | mutant enzyme H144L/W187I, at pH 6.5 and 30°C | Alcaligenes faecalis | |
36.83 | - |
levulinic acid | mutant enzyme W187A, at pH 6.5 and 30°C | Alcaligenes faecalis | |
42.1 | - |
levulinic acid | mutant enzyme W187V, 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 | - |
- |
Purification (Comment) | Organism |
---|---|
Ni-NTA column chromatography | Alcaligenes faecalis |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
acetoacetate + NADH + H+ | - |
Alcaligenes faecalis | (R)-3-hydroxybutanoate + NAD+ | - |
? | |
levulinic acid + NADH + H+ | the wild type enzyme shows low activity with levulinic acid, while it's a good substrate for mutant enzyme H144L/W187F | 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 |
---|---|---|---|---|---|
0.00016 | - |
levulinic acid | mutant enzyme W187A, at pH 6.5 and 30°C | Alcaligenes faecalis | |
0.0003 | - |
levulinic acid | mutant enzyme W187I, at pH 6.5 and 30°C | Alcaligenes faecalis | |
0.0003 | - |
levulinic acid | mutant enzyme W187V, at pH 6.5 and 30°C | Alcaligenes faecalis | |
0.00084 | - |
levulinic acid | mutant enzyme H144A, at pH 6.5 and 30°C | Alcaligenes faecalis | |
0.00122 | - |
levulinic acid | mutant enzyme H144L/W187A, at pH 6.5 and 30°C | Alcaligenes faecalis | |
0.002 | - |
levulinic acid | mutant enzyme H144L/W187V, at pH 6.5 and 30°C | Alcaligenes faecalis | |
0.00289 | - |
levulinic acid | mutant enzyme H144L/W187I, at pH 6.5 and 30°C | Alcaligenes faecalis | |
0.00465 | - |
levulinic acid | mutant enzyme W187L, at pH 6.5 and 30°C | Alcaligenes faecalis | |
0.00766 | - |
levulinic acid | mutant enzyme H144V, at pH 6.5 and 30°C | Alcaligenes faecalis | |
0.00768 | - |
levulinic acid | wild type enzyme, at pH 6.5 and 30°C | Alcaligenes faecalis | |
0.00921 | - |
levulinic acid | mutant enzyme H144F, at pH 6.5 and 30°C | Alcaligenes faecalis | |
0.01069 | - |
levulinic acid | mutant enzyme H144I, at pH 6.5 and 30°C | Alcaligenes faecalis | |
0.02969 | - |
levulinic acid | mutant enzyme W187F, at pH 6.5 and 30°C | Alcaligenes faecalis | |
0.06594 | - |
levulinic acid | mutant enzyme H144L, at pH 6.5 and 30°C | Alcaligenes faecalis | |
0.07107 | - |
levulinic acid | mutant enzyme H144L/W187L, at pH 6.5 and 30°C | Alcaligenes faecalis | |
0.2565 | - |
levulinic acid | mutant enzyme H144L/W187F, 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 |
---|---|---|---|---|---|
0.006 | - |
levulinic acid | wild type enzyme, at pH 6.5 and 30°C | Alcaligenes faecalis | |
0.006 | - |
levulinic acid | mutant enzyme W187A, at pH 6.5 and 30°C | Alcaligenes faecalis | |
0.008 | - |
levulinic acid | mutant enzyme H144A, at pH 6.5 and 30°C | Alcaligenes faecalis | |
0.013 | - |
levulinic acid | mutant enzyme W187V, at pH 6.5 and 30°C | Alcaligenes faecalis | |
0.014 | - |
levulinic acid | mutant enzyme W187I, at pH 6.5 and 30°C | Alcaligenes faecalis | |
0.016 | - |
levulinic acid | mutant enzyme H144F, at pH 6.5 and 30°C | Alcaligenes faecalis | |
0.016 | - |
levulinic acid | mutant enzyme H144L/W187A, at pH 6.5 and 30°C | Alcaligenes faecalis | |
0.019 | - |
levulinic acid | mutant enzyme W187F, at pH 6.5 and 30°C | Alcaligenes faecalis | |
0.03 | - |
levulinic acid | mutant enzyme H144I, at pH 6.5 and 30°C | Alcaligenes faecalis | |
0.03 | - |
levulinic acid | mutant enzyme H144V, at pH 6.5 and 30°C | Alcaligenes faecalis | |
0.045 | - |
levulinic acid | mutant enzyme H144L/W187L, at pH 6.5 and 30°C | Alcaligenes faecalis | |
0.051 | - |
levulinic acid | mutant enzyme H144L/W187V, at pH 6.5 and 30°C | Alcaligenes faecalis | |
0.06 | - |
levulinic acid | mutant enzyme H144L, at pH 6.5 and 30°C | Alcaligenes faecalis | |
0.063 | - |
levulinic acid | mutant enzyme W187L, at pH 6.5 and 30°C | Alcaligenes faecalis | |
0.074 | - |
levulinic acid | mutant enzyme H144L/W187I, at pH 6.5 and 30°C | Alcaligenes faecalis | |
0.164 | - |
levulinic acid | mutant enzyme H144L/W187F, at pH 6.5 and 30°C | Alcaligenes faecalis |