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Literature summary extracted from

  • Chen, D.; Tang, H.; Lv, Y.; Zhang, Z.; Shen, K.; Lin, K.; Zhao, Y.L.; Wu, G.; Xu, P.
    Structural and computational studies of the maleate isomerase from Pseudomonas putida S16 reveal a breathing motion wrapping the substrate inside (2013), Mol. Microbiol., 87, 1237-1244.
    View publication on PubMed

Cloned(Commentary)

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

Crystallization (Commentary)

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

Protein Variants

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

Natural Substrates/ Products (Substrates)

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

Organism

EC Number Organism UniProt Comment Textmining
5.2.1.1 Pseudomonas putida F8G0M3
-
-
5.2.1.1 Pseudomonas putida S16 F8G0M3
-
-

Purification (Commentary)

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

Substrates and Products (Substrate)

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

Synonyms

EC Number Synonyms Comment Organism
5.2.1.1 Pp-Iso
-
 Pseudomonas putida

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
5.2.1.1 30
-
 assay at Pseudomonas putida

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
5.2.1.1 8.4
-
 assay at Pseudomonas putida

General Information

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