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Literature summary extracted from

  • Drevland, R.M.; Jia, Y.; Palmer, D.R.; Graham, D.E.
    Methanogen homoaconitase catalyzes both hydrolyase reactions in coenzyme B biosynthesis (2008), J. Biol. Chem., 283, 28888-28896.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
4.2.1.114 DNA and amino acid sequence determination and analysis, coexpression of MJ1003 and MJ1271 proteins in Escherichia coli strain BL21(DE3) Methanocaldococcus jannaschii

Inhibitors

EC Number Inhibitors Comment Organism Structure
4.2.1.114 (S)-homocitrate complete inhibition at 0.1 mM Methanocaldococcus jannaschii
4.2.1.114 cis-aconitate 50% inhibition of the hydrolase activity at 0.025 mM Methanocaldococcus jannaschii
4.2.1.114 cis-homoaconitate/(R)-homocitrate mixtures of 0.1 mM cis-homoaconitate and 0.05 mM (R)-homocitrate reduce the HACN hydrolyase activity by 50% Methanocaldococcus jannaschii
4.2.1.114 additional information no inhibition by citraconate up to 0.2 mM Methanocaldococcus jannaschii
4.2.1.114 trans-(homo)3aconitate 50% inhibition of the hydrolase activity at 0.4 mM Methanocaldococcus jannaschii
4.2.1.114 trans-aconitate 50% inhibition of the hydrolase activity at 0.1 mM Methanocaldococcus jannaschii

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
4.2.1.114 additional information
-
 additional information steady-state kinetics Methanocaldococcus jannaschii
4.2.1.114 0.022
-
 cis-homoaconitate pH 9.0, 60°C, recombinant enzyme, hydrolase reaction Methanocaldococcus jannaschii
4.2.1.114 0.03
-
 cis-homo2aconitate pH 9.0, 60°C, recombinant enzyme, hydrolase reaction Methanocaldococcus jannaschii
4.2.1.114 0.036
-
 cis-homo3aconitate pH 9.0, 60°C, recombinant enzyme, hydrolase reaction Methanocaldococcus jannaschii
4.2.1.114 0.175
-
 cis-homo4aconitate pH 9.0, 60°C, recombinant enzyme, hydrolase reaction Methanocaldococcus jannaschii
4.2.1.114 0.33
-
 Maleate pH 9.0, 60°C, recombinant enzyme, hydrolase reaction Methanocaldococcus jannaschii
4.2.1.114 1.5
-
 (R)-homocitrate pH 8.5, 60°C, recombinant enzyme, dehydratase reaction Methanocaldococcus jannaschii

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
4.2.1.114 Fe2+ the enzyme contains an iron-sulfur center, reconstitution of the purified recombinant apoenzyme with iron-sulfur, overview Methanocaldococcus jannaschii
4.2.1.114 Mg2+ required for the hydratase reaction, not for the hydrolase reaction Methanocaldococcus jannaschii

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
4.2.1.114 cis-(homo)2aconitate + H2O Methanocaldococcus jannaschii
-
 (-)-threo-iso(homo)2citrate
-
 ?
4.2.1.114 cis-(homo)3aconitate + H2O Methanocaldococcus jannaschii
-
 (-)-threo-iso(homo)3citrate
-
 ?
4.2.1.114 additional information Methanocaldococcus jannaschii homoaconitase enzymes catalyze hydrolyase reactions in the alpha-aminoadipate pathway for lysine biosynthesis or the 2-oxosuberate pathway for methanogenic coenzyme B biosynthesis ?
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
4.2.1.114 Methanocaldococcus jannaschii
-
-
-
4.2.1.114 Methanocaldococcus jannaschii Q58991 the sequence contains the alteration C229A resulting in Q77K amino acid exchange compared to the MJ1596 sequence reported by the genome sequencing project, cf. EC 1.1.1.87; proteins MJ1003 and MJ1271
-

Purification (Commentary)

EC Number Purification (Comment) Organism
4.2.1.114 recombinant MJ1003 and MJ1271 proteins from Escherichia coli strain BL21(DE3) by heat treatment of the cell lysate, followed by anion exchange chromatography, dialysis, and ultrafiltration Methanocaldococcus jannaschii

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4.2.1.114 (R)-2-hydroxybutane-1,2,4-tricarboxylate i.e. (R)-homocitrate, overall reaction, the enzyme catalyzes both the dehydration of (R)-homocitrate to form cis-homoaconitate, and hydration producing homoisocitrate Methanocaldococcus jannaschii (1R,2S)-1-hydroxybutane-1,2,4-tricarboxylate i.e. homoisocitrate r
4.2.1.114 (R)-2-hydroxybutane-1,2,4-tricarboxylate i.e. (R)-homocitrate, first half-reaction Methanocaldococcus jannaschii (Z)-but-1-ene-1,2,4-tricarboxylate + H2O i.e. cis-homoaconitate, the activity in the reverse direction is similar as with (R)-homocitrate r
4.2.1.114 (Z)-but-1-ene-1,2,4-tricarboxylate + H2O second half-reaction Methanocaldococcus jannaschii (1R,2S)-1-hydroxybutane-1,2,4-tricarboxylate i.e. homoisocitrate r
4.2.1.114 cis-(homo)2aconitate + H2O the activity is similar as with (R)-homocitrate Methanocaldococcus jannaschii ?
-
 ?
4.2.1.114 cis-(homo)2aconitate + H2O
-
 Methanocaldococcus jannaschii (-)-threo-iso(homo)2citrate
-
 ?
4.2.1.114 cis-(homo)3aconitate + H2O the activity is similar as with (R)-homocitrate Methanocaldococcus jannaschii ?
-
 ?
4.2.1.114 cis-(homo)3aconitate + H2O
-
 Methanocaldococcus jannaschii (-)-threo-iso(homo)3citrate
-
 ?
4.2.1.114 cis-(homo)4aconitate + H2O the activity is similar as with (R)-homocitrate Methanocaldococcus jannaschii ?
-
 ?
4.2.1.114 maleate + H2O
-
 Methanocaldococcus jannaschii ?
-
 ?
4.2.1.114 additional information homoaconitase enzymes catalyze hydrolyase reactions in the alpha-aminoadipate pathway for lysine biosynthesis or the 2-oxosuberate pathway for methanogenic coenzyme B biosynthesis Methanocaldococcus jannaschii ?
-
 ?
4.2.1.114 additional information homoaconitase together with the endogenous homoisocitrate dehydrogenase, EC 1.1.1.87, catalyze all of the isomerization and oxidative decarboxylation reactions required to form 2-oxoadipate, 2-oxopimelate, and 2-oxosuberate, completing three iterations of the 2-oxoacid elongation pathway. Methanogenic archaeal homoaconitases and fungal homoaconitases evolved in parallel in the aconitase superfamily, LC-MS analysis of enzyme reaction products, overview Methanocaldococcus jannaschii ?
-
 ?

Subunits

EC Number Subunits Comment Organism
4.2.1.114 tetramer heterotetramer, MJ1003 and MJ1271 proteins form an active homoaconitase enzyme Methanocaldococcus jannaschii

Synonyms

EC Number Synonyms Comment Organism
4.2.1.114 HACN
-
 Methanocaldococcus jannaschii
4.2.1.114 Homoaconitase
-
 Methanocaldococcus jannaschii
4.2.1.114 More the enzyme belongs to the aconitase superfamily Methanocaldococcus jannaschii

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
4.2.1.114 60
-
 hydrolase and hydratase assay at Methanocaldococcus jannaschii

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
4.2.1.114 0.37
-
 (R)-homocitrate pH 8.5, 60°C, recombinant enzyme, dehydratase reaction Methanocaldococcus jannaschii
4.2.1.114 0.66
-
 cis-homo2aconitate pH 9.0, 60°C, recombinant enzyme, hydrolase reaction Methanocaldococcus jannaschii
4.2.1.114 0.75
-
 cis-homoaconitate pH 9.0, 60°C, recombinant enzyme, hydrolase reaction Methanocaldococcus jannaschii
4.2.1.114 2.5
-
 cis-homo3aconitate pH 9.0, 60°C, recombinant enzyme, hydrolase reaction Methanocaldococcus jannaschii
4.2.1.114 5.6
-
 cis-homo4aconitate pH 9.0, 60°C, recombinant enzyme, hydrolase reaction Methanocaldococcus jannaschii
4.2.1.114 6
-
 Maleate pH 9.0, 60°C, recombinant enzyme, hydrolase reaction Methanocaldococcus jannaschii

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
4.2.1.114 8.5
-
 hydratase assay at Methanocaldococcus jannaschii
4.2.1.114 9
-
 hydrolase assay at Methanocaldococcus jannaschii