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Literature summary extracted from

  • Campos-Garcia, J.; Diaz-Perez, C.; Diaz-Perez, A.L.
    Residues Asn214, Gln211, Glu219 and Gln221 contained in the subfamily 3 catalytic signature of the isocitrate lyase from Pseudomonas aeruginosa are involved in its catalytic and thermal properties (2013), World J. Microbiol. Biotechnol., 29, 991-999.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
4.1.3.1 gene aceA, phylogenetic analysis, expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) Pseudomonas aeruginosa

Protein Variants

EC Number Protein Variants Comment Organism
4.1.3.1 E219A site-directed mutagenesis of the highly conserved residue in the ICL subfamily 3 catalytic pattern, the mutant enzyme shows reduced activity compared to the wild-type enzyme Pseudomonas aeruginosa
4.1.3.1 N214D site-directed mutagenesis of the highly conserved residue in the ICL subfamily 3 catalytic pattern, the mutant enzyme shows 10fold reduced activity compared to the wild-type enzyme Pseudomonas aeruginosa
4.1.3.1 Q211H site-directed mutagenesis of the highly conserved residue in the ICL subfamily 3 catalytic pattern, the mutant enzyme shows reduced activity and elevated temperature optimum compared to the wild-type enzyme Pseudomonas aeruginosa
4.1.3.1 Q221K site-directed mutagenesis of the highly conserved residue in the ICL subfamily 3 catalytic pattern, the mutant enzyme shows reduced activity and elevated temperature optimum compared to the wild-type enzyme Pseudomonas aeruginosa

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
4.1.3.1 additional information
-
 additional information Michaelis-Menten kinetics of wild-type and mutant enzyme's overview Pseudomonas aeruginosa
4.1.3.1 0.25
-
 isocitrate recombinant His-tagged mutant N214D, pH 7.8, 30°C Pseudomonas aeruginosa
4.1.3.1 0.29
-
 isocitrate recombinant His-tagged mutant Q211H, pH 7.8, 30°C Pseudomonas aeruginosa
4.1.3.1 0.44
-
 isocitrate recombinant His-tagged wild-type enzyme, pH 7.8, 30°C Pseudomonas aeruginosa
4.1.3.1 0.47
-
 isocitrate recombinant His-tagged mutant Q221K, pH 7.8, 30°C Pseudomonas aeruginosa
4.1.3.1 0.94
-
 isocitrate recombinant His-tagged mutant E219A, pH 7.8, 30°C Pseudomonas aeruginosa

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
4.1.3.1 isocitrate Pseudomonas aeruginosa
-
 succinate + glyoxylate
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
4.1.3.1 Pseudomonas aeruginosa Q9I0K4 gene aceA
-

Purification (Commentary)

EC Number Purification (Comment) Organism
4.1.3.1 ecombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography Pseudomonas aeruginosa

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4.1.3.1 isocitrate
-
 Pseudomonas aeruginosa succinate + glyoxylate
-
 ?

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
4.1.3.1 30
-
 wild-type enzyme and mutant E219A Pseudomonas aeruginosa
4.1.3.1 36
-
 mutant Q221K Pseudomonas aeruginosa
4.1.3.1 39
-
 mutant Q211H Pseudomonas aeruginosa

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
4.1.3.1 45
-
 purified enzyme, half-life of the wild-type enzyme is 40 min, half-lives of mutants E219A, Q211H, and Q221K are 25 min, 12 min, and 15 min Pseudomonas aeruginosa

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
4.1.3.1 0.56
-
 isocitrate recombinant His-tagged mutant N214D, pH 7.8, 30°C Pseudomonas aeruginosa
4.1.3.1 3.25
-
 isocitrate recombinant His-tagged mutant Q221K, pH 7.8, 30°C Pseudomonas aeruginosa
4.1.3.1 3.46
-
 isocitrate recombinant His-tagged mutant Q211H, pH 7.8, 30°C Pseudomonas aeruginosa
4.1.3.1 4.63
-
 isocitrate recombinant His-tagged mutant E219A, pH 7.8, 30°C Pseudomonas aeruginosa
4.1.3.1 9.81
-
 isocitrate recombinant His-tagged wild-type enzyme, pH 7.8, 30°C Pseudomonas aeruginosa

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
4.1.3.1 7.6
-
-
 Pseudomonas aeruginosa

General Information

EC Number General Information Comment Organism
4.1.3.1 evolution the ICL family includes five subfamilies, and the 2-methylisocitrate lyase (MICL) family. The ICL from Pseudomonas aeruginosa (ICL-Pa) is identified in a different ICL node (subfamily 3) than other Pseudomonas ICL enzymes, phylogenetic analysis Pseudomonas aeruginosa
4.1.3.1 additional information conserved residues in the subfamily 3 signature of ICL-Pa play important roles in catalysis and thermostability and are likely associated with the catalytic loop structural conformation. Three-dimensional structural homology modeling of ICL-Pa wild-type and mutants, structure comparisons, active site modeling, overview. Residue N214 plays an essential role in ICL-Pa catalytic activity Pseudomonas aeruginosa
4.1.3.1 physiological function isocitrate lyase plays an important role in the ability of Pseudomonas aeruginosa to grow on fatty acids, acetate, acyclic terpenes, and amino acids Pseudomonas aeruginosa

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
4.1.3.1 2.8
-
 isocitrate recombinant His-tagged mutant N214D, pH 7.8, 30°C Pseudomonas aeruginosa
4.1.3.1 4.9
-
 isocitrate recombinant His-tagged mutant E219A, pH 7.8, 30°C Pseudomonas aeruginosa
4.1.3.1 6.9
-
 isocitrate recombinant His-tagged mutant Q221K, pH 7.8, 30°C Pseudomonas aeruginosa
4.1.3.1 11.9
-
 isocitrate recombinant His-tagged mutant Q211H, pH 7.8, 30°C Pseudomonas aeruginosa
4.1.3.1 22.3
-
 isocitrate recombinant His-tagged wild-type enzyme, pH 7.8, 30°C Pseudomonas aeruginosa