EC Number | Cloned (Comment) | Organism |
---|---|---|
5.2.1.1 | overexpression of N- and C-terminally His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) | Pseudomonas putida |
EC Number | Crystallization (Comment) | Organism |
---|---|---|
5.2.1.1 | purified recombinant wild-type alone, and mutant enzyme C200A in complex with substrate maleate, hanging-drop vapour-diffusion method, mixing of 15-20 mg/ml protein in 25 mM Tris-HCl, pH 8.0, 150 mM NaCl, and 2 mM DTT, with a reservoir solution containing 25% w/v PEG3,350 and 0.1 M Tris, pH 8.5, for the substrate-enzyme mutant complex, and 1.4 M (NH4)2SO4, 120 mM NaCl, and 0.1 M HEPES, pH 7.5, for the free wild-type enzyme, 14°C, X-ray diffraction structure determination and analysis at 2.95 A and 2.10 A resolution, respectively | Pseudomonas putida |
EC Number | Protein Variants | Comment | Organism |
---|---|---|---|
5.2.1.1 | C200A | site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme, substrate-binding structure: maleate can be found to exist inside a cavity of the enzyme, totally surrounded by the protein | Pseudomonas putida |
5.2.1.1 | C84A | site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type enzyme | Pseudomonas putida |
EC Number | Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
5.2.1.1 | Maleate | Pseudomonas putida | - |
Fumarate | - |
r | |
5.2.1.1 | Maleate | Pseudomonas putida S16 | - |
Fumarate | - |
r |
EC Number | Organism | UniProt | Comment | Textmining |
---|---|---|---|---|
5.2.1.1 | Pseudomonas putida | F8G0M3 | - |
- |
5.2.1.1 | Pseudomonas putida S16 | F8G0M3 | - |
- |
EC Number | Purification (Comment) | Organism |
---|---|---|
5.2.1.1 | recombinant N- and C-terminally His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and gel filtration | Pseudomonas putida |
EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
5.2.1.1 | Maleate | - |
Pseudomonas putida | Fumarate | - |
r | |
5.2.1.1 | Maleate | Asn17 and Asn169 play critical roles in recognizing maleate | Pseudomonas putida | Fumarate | - |
r | |
5.2.1.1 | Maleate | - |
Pseudomonas putida S16 | Fumarate | - |
r | |
5.2.1.1 | Maleate | Asn17 and Asn169 play critical roles in recognizing maleate | Pseudomonas putida S16 | Fumarate | - |
r |
EC Number | Synonyms | Comment | Organism |
---|---|---|---|
5.2.1.1 | Pp-Iso | - |
Pseudomonas putida |
EC Number | Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|---|
5.2.1.1 | 30 | - |
assay at | Pseudomonas putida |
EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|---|
5.2.1.1 | 8.4 | - |
assay at | Pseudomonas putida |
EC Number | General Information | Comment | Organism |
---|---|---|---|
5.2.1.1 | metabolism | maleate isomerase catalyses the last step in nicotine degradation, the cis-trans isomerization of maleate to fumarate | Pseudomonas putida |
5.2.1.1 | additional information | residues C84 and C200 are essential for catalysis, active site structure, overview. The beta2-alpha2 loop and the beta6-alpha7 loop of Pp-Iso have a breathing motion revealed by the molecular dynamics simulation | Pseudomonas putida |