Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli BL21(DE3) cells | Mycolicibacterium smegmatis |
Protein Variants | Comment | Organism |
---|---|---|
P1A | for phenylenolpyruvate, the P1A mutant shows a 41fold decrease in kcat and a 14fold decrease in Km, resulting in an about 3fold decrease in kcat/Km. For (4-hydroxyphenyl)enolpyruvate, the P1A mutant shows a 16fold decrease in kcat, whereas the Km is not significantly affected, resulting in a 16fold decrease in kcat/Km. The P1A mutant has no detectable dehalogenase activity toward cis-3-chloroacrylate and trans-3-chloroacrylate, and no detectable activity toward 2-oxo-3-pentynoate (at pH 7.3) | Mycolicibacterium smegmatis |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
2-Oxo-3-pentynoate | at pH 8.5, CCH2 is inactivated by 2-oxo-3-pentynoate due to the covalent modification of Pro-1 | Mycolicibacterium smegmatis |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.11 | - |
phenylenolpyruvate | wild type enzyme cis-CAAD, in 10 mM Na2HPO4 buffer, pH 7.3, at 22°C | Mycolicibacterium smegmatis | |
0.152 | - |
cis-3-chloroacrylate | wild type enzyme cis-CAAD, in 10 mM Na2HPO4 buffer, pH 7.3, at 22°C | Mycolicibacterium smegmatis | |
0.25 | - |
phenylenolpyruvate | mutant enzyme CCH2-P1A, in 10 mM Na2HPO4 buffer, pH 7.3, at 22°C | Mycolicibacterium smegmatis | |
0.79 | - |
(4-hydroxyphenyl)enolpyruvate | wild type enzyme CCH2-P1A, in 10 mM Na2HPO4 buffer, pH 7.3, at 22°C | Mycolicibacterium smegmatis | |
0.89 | - |
(4-hydroxyphenyl)enolpyruvate | wild type enzyme CCH2, in 10 mM Na2HPO4 buffer, pH 7.3, at 22°C | Mycolicibacterium smegmatis | |
3.5 | - |
phenylenolpyruvate | wild type enzyme CCH2, in 10 mM Na2HPO4 buffer, pH 7.3, at 22°C | Mycolicibacterium smegmatis | |
96 | - |
trans-3-chloroacrylate | wild type enzyme CCH2, in in 50 mM Tris-SO4 buffer (pH 8.0), at 22°C | Mycolicibacterium smegmatis |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
16093 | - |
3 * 16093, electrospray ionization mass spectrometry | Mycolicibacterium smegmatis |
16223 | - |
3 * 16223, calculated from amino acid sequence | Mycolicibacterium smegmatis |
48000 | - |
gel filtration | Mycolicibacterium smegmatis |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Mycolicibacterium smegmatis | - |
- |
- |
Mycolicibacterium smegmatis mc(2)155 | - |
- |
- |
Purification (Comment) | Organism |
---|---|
Ni-Sepharose column chromatography | Mycolicibacterium smegmatis |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
(4-hydroxyphenyl)enolpyruvate | - |
Mycolicibacterium smegmatis | (4-hydroxyphenyl)pyruvate | - |
? | |
(4-hydroxyphenyl)enolpyruvate | - |
Mycolicibacterium smegmatis mc(2)155 | (4-hydroxyphenyl)pyruvate | - |
? | |
2-oxo-3-pentynoate | at pH 6.5, CCH2 exhibits hydratase activity and converts 2-oxo-3-pentynoate to acetopyruvate | Mycolicibacterium smegmatis | acetopyruvate + ? | - |
? | |
2-oxo-3-pentynoate | at pH 6.5, CCH2 exhibits hydratase activity and converts 2-oxo-3-pentynoate to acetopyruvate | Mycolicibacterium smegmatis mc(2)155 | acetopyruvate + ? | - |
? | |
cis-3-chloroacrylate | the reaction is performed by cis-CAAD | Mycolicibacterium smegmatis | malonate semialdehyde + HCl | - |
? | |
cis-3-chloroacrylate | the reaction is performed by cis-CAAD | Mycolicibacterium smegmatis mc(2)155 | malonate semialdehyde + HCl | - |
? | |
additional information | CCH2 functions as an efficient PPT and exhibits low-level promiscuous dehalogenase activity, processing both cis- and trans-3-chloroacrylic acid | Mycolicibacterium smegmatis | ? | - |
? | |
additional information | CCH2 functions as an efficient PPT and exhibits low-level promiscuous dehalogenase activity, processing both cis- and trans-3-chloroacrylic acid | Mycolicibacterium smegmatis mc(2)155 | ? | - |
? | |
phenylenolpyruvate | - |
Mycolicibacterium smegmatis | phenylpyruvate | - |
? | |
phenylenolpyruvate | - |
Mycolicibacterium smegmatis mc(2)155 | phenylpyruvate | - |
? | |
trans-3-chloroacrylate | the reaction is performed by CAAD | Mycolicibacterium smegmatis | malonate semialdehyde + HCl | - |
? |
Subunits | Comment | Organism |
---|---|---|
homotrimer | 3 * 16093, electrospray ionization mass spectrometry | Mycolicibacterium smegmatis |
homotrimer | 3 * 16223, calculated from amino acid sequence | Mycolicibacterium smegmatis |
Synonyms | Comment | Organism |
---|---|---|
CaaD | - |
Mycolicibacterium smegmatis |
CCH2 | - |
Mycolicibacterium smegmatis |
cis-3-chloroacrylic acid dehalogenase | - |
Mycolicibacterium smegmatis |
cis-CaaD | - |
Mycolicibacterium smegmatis |
phenyl(enol)pyruvate tautomerase | - |
Mycolicibacterium smegmatis |
PPT | - |
Mycolicibacterium smegmatis |
trans-3-chloroacrylic acid dehalogenase | - |
Mycolicibacterium smegmatis |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0004 | - |
trans-3-chloroacrylate | wild type enzyme CCH2, in in 50 mM Tris-SO4 buffer (pH 8.0), at 22°C | Mycolicibacterium smegmatis | |
0.2 | - |
(4-hydroxyphenyl)enolpyruvate | wild type enzyme CCH2-P1A, in 10 mM Na2HPO4 buffer, pH 7.3, at 22°C | Mycolicibacterium smegmatis | |
0.2 | - |
phenylenolpyruvate | wild type enzyme cis-CAAD, in 10 mM Na2HPO4 buffer, pH 7.3, at 22°C | Mycolicibacterium smegmatis | |
0.8 | - |
phenylenolpyruvate | mutant enzyme CCH2-P1A, in 10 mM Na2HPO4 buffer, pH 7.3, at 22°C | Mycolicibacterium smegmatis | |
3.2 | - |
(4-hydroxyphenyl)enolpyruvate | wild type enzyme CCH2, in 10 mM Na2HPO4 buffer, pH 7.3, at 22°C | Mycolicibacterium smegmatis | |
4.6 | - |
cis-3-chloroacrylate | wild type enzyme cis-CAAD, in 10 mM Na2HPO4 buffer, pH 7.3, at 22°C | Mycolicibacterium smegmatis | |
34 | - |
phenylenolpyruvate | wild type enzyme CCH2, in 10 mM Na2HPO4 buffer, pH 7.3, at 22°C | Mycolicibacterium smegmatis |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
1.6 | - |
cis-3-chloroacrylate | wild type enzyme CCH2, in in 50 mM Tris-SO4 buffer (pH 8.0), at 22°C | Mycolicibacterium smegmatis | |
1.8 | - |
phenylenolpyruvate | wild type enzyme cis-CAAD, in 10 mM Na2HPO4 buffer, pH 7.3, at 22°C | Mycolicibacterium smegmatis | |
2.5 | - |
(4-hydroxyphenyl)enolpyruvate | wild type enzyme CCH2-P1A, in 10 mM Na2HPO4 buffer, pH 7.3, at 22°C | Mycolicibacterium smegmatis | |
3.3 | - |
phenylenolpyruvate | mutant enzyme CCH2-P1A, in 10 mM Na2HPO4 buffer, pH 7.3, at 22°C | Mycolicibacterium smegmatis | |
3.5 | - |
(4-hydroxyphenyl)enolpyruvate | wild type enzyme CCH2, in 10 mM Na2HPO4 buffer, pH 7.3, at 22°C | Mycolicibacterium smegmatis | |
4 | - |
trans-3-chloroacrylate | wild type enzyme CCH2, in in 50 mM Tris-SO4 buffer (pH 8.0), at 22°C | Mycolicibacterium smegmatis | |
9.8 | - |
phenylenolpyruvate | wild type enzyme CCH2, in 10 mM Na2HPO4 buffer, pH 7.3, at 22°C | Mycolicibacterium smegmatis | |
30 | - |
cis-3-chloroacrylate | wild type enzyme cis-CAAD, in 10 mM Na2HPO4 buffer, pH 7.3, at 22°C | Mycolicibacterium smegmatis |