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Literature summary extracted from

  • Okada, T.; Tomita, T.; Wulandari, A.P.; Kuzuyama, T.; Nishiyama, M.
    Mechanism of substrate recognition and insight into feedback inhibition of homocitrate synthase from Thermus thermophilus (2010), J. Biol. Chem., 285, 4195-4205.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.3.3.14 three expression systems are designed, TtHCS without tag, with a C-terminal His6 tag and with a N-terminal His8 tag, the vectors pBluescript II SK+ and pET26b+ are used for expression in Escherichia coli cells Thermus thermophilus

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
2.3.3.14 the crystal structures of homocitrate synthase complexed with 2-oxoglutarate, (R)-2-hydroxybutane-1,2,4-tricarboxylate, or lysine are determined Thermus thermophilus

Protein Variants

EC Number Protein Variants Comment Organism
2.3.3.14 H72L replacement of His72 by leucine makes HCS resistant to lysine inhibition, demonstrating the regulatory role of this conserved residue Thermus thermophilus

Inhibitors

EC Number Inhibitors Comment Organism Structure
2.3.3.14 lysine regulated via feedback inhibition by the endproduct Thermus thermophilus

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.3.3.14 0.0054
-
 2-oxoglutarate wild-type enzyme, N-terminal His8 tag Thermus thermophilus
2.3.3.14 0.033
-
 acetyl-CoA wild-type enzyme, N-terminal His8 tag Thermus thermophilus
2.3.3.14 0.037
-
 acetyl-CoA H72L mutant, N-terminal His8 tag Thermus thermophilus
2.3.3.14 0.174
-
 2-oxoglutarate H72L mutant, N-terminal His8 tag Thermus thermophilus

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.3.3.14 acetyl-CoA + H2O + 2-oxoglutarate Thermus thermophilus
-
 (R)-2-hydroxybutane-1,2,4-tricarboxylate + CoA
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
2.3.3.14 Thermus thermophilus
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.3.3.14 a Ni2+-nitrilotriacetic acid column, a gel filtration column, a HiLoad 26/60 phenyl-Sepharose column and a Resource Q anion exchange column are used Thermus thermophilus

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
2.3.3.14
-
-
 no activity without additive Thermus thermophilus
2.3.3.14 0.0003
-
 additive CuCl2 Thermus thermophilus
2.3.3.14 0.0008
-
 additive FeSO4 Thermus thermophilus
2.3.3.14 0.0013
-
 additive ZnSO4 Thermus thermophilus
2.3.3.14 0.0036
-
 additive NiCl2 Thermus thermophilus
2.3.3.14 0.0075
-
 additive CaCl2 Thermus thermophilus
2.3.3.14 0.0333
-
 additive CoCl2 Thermus thermophilus
2.3.3.14 0.0392
-
 additive MnCl2 Thermus thermophilus
2.3.3.14 0.0415
-
 additive MgSO4 Thermus thermophilus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.3.3.14 acetyl-CoA + H2O + 2-oxoglutarate
-
 Thermus thermophilus (R)-2-hydroxybutane-1,2,4-tricarboxylate + CoA
-
 ?

Synonyms

EC Number Synonyms Comment Organism
2.3.3.14 HCS
-
 Thermus thermophilus
2.3.3.14 homocitrate synthase
-
 Thermus thermophilus
2.3.3.14 TtHCS
-
 Thermus thermophilus

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.3.3.14 50
-
 activity assay Thermus thermophilus

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.3.3.14 0.182
-
 2-oxoglutarate wild-type enzyme, N-terminal His8 tag Thermus thermophilus
2.3.3.14 0.248
-
 2-oxoglutarate H72L mutant, N-terminal His8 tag Thermus thermophilus
2.3.3.14 0.517
-
 acetyl-CoA H72L mutant, N-terminal His8 tag Thermus thermophilus
2.3.3.14 0.683
-
 acetyl-CoA wild-type enzyme, N-terminal His8 tag Thermus thermophilus

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.3.3.14 7.5
-
 activity assay Thermus thermophilus

Cofactor

EC Number Cofactor Comment Organism Structure
2.3.3.14 acetyl-CoA
-
 Thermus thermophilus

Ki Value [mM]

EC Number Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
2.3.3.14 0.0707
-
 lysine wild-type enzyme, N-terminal His8 tag Thermus thermophilus
2.3.3.14 2
-
 lysine H72L mutant, N-terminal His8 tag, larger than 2 Thermus thermophilus

General Information

EC Number General Information Comment Organism
2.3.3.14 metabolism reaction is the first step in the lysine biosynthetic pathway through alpha-aminoadipate Thermus thermophilus