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Literature summary extracted from

  • Nagegowda, D.A.; Bach, T.J.; Chye, M.L.
    Brassica juncea 3-hydroxy-3-methylglutaryl (HMG)-CoA synthase 1: expression and characterization of recombinant wild-type and mutant enzymes (2004), Biochem. J., 383, 517-527.
    View publication on PubMedView publication on EuropePMC

Activating Compound

EC Number Activating Compound Comment Organism Structure
2.3.3.10 dithioerythritol 5 mM, activates Brassica juncea

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.3.3.10 His6-tagged protein from Escherichia coli,His6-BjHMGS1 Brassica juncea

Protein Variants

EC Number Protein Variants Comment Organism
2.3.3.10 C212C mutation causes 6fold decreased maximal velocity Brassica juncea
2.3.3.10 H188N mutation causes 8fold decreased maximal velocity, mutant enzyme does not display substrate inhibition by acetoacetyl-CoA Brassica juncea
2.3.3.10 H188N/S359A 10fold increased specific activity, no inhibition by acetoacetyl-CoA Brassica juncea
2.3.3.10 N115A KI value for acetoacetyl-CoA is 2.3fold higher than the wild-type value, the turnover number for acetyl-CoA is 17.7fold lower than wild-type value, KM-value is 1.6fold lower than wild-type value Brassica juncea
2.3.3.10 R157A mutation causes 14fold decreased maximal velocity Brassica juncea
2.3.3.10 S356A KI value for acetoacetyl-CoA is 5.2fold higher than the wild-type value, the turnover number for acetyl-CoA is 25.8fold lower than wild-type value, KM-value is 1.6fold lower than wild-type value Brassica juncea
2.3.3.10 S359A 10fold increased specific activity Brassica juncea
2.3.3.10 S89A KI value for acetoacetyl-CoA is 1.2fold higher than the wild-type value, the turnover number for acetyl-CoA is 54.6fold lower than wild-type value, KM-value is 2.3fold lower than wild-type value Brassica juncea

Inhibitors

EC Number Inhibitors Comment Organism Structure
2.3.3.10 (S)-3-hydroxy-3-methylglutaryl-CoA
-
 Brassica juncea
2.3.3.10 acetoacetyl-CoA inhibition of wild-type enzyme, no inhibition of mutant enzyme H188N and H188N/S259A Brassica juncea
2.3.3.10 Co2+
-
 Brassica juncea
2.3.3.10 CoA
-
 Brassica juncea
2.3.3.10 F-244 IC50: 35 nM Brassica juncea
2.3.3.10 Mn2+
-
 Brassica juncea
2.3.3.10 Zn2+
-
 Brassica juncea

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.3.3.10 0.0029
-
 acetoacetyl-CoA pH 7.5, 30°C, mutant enzyme H188N/S359A Brassica juncea
2.3.3.10 0.012
-
 acetoacetyl-CoA pH 7.5, 30°C, mutant enzyme H188N Brassica juncea
2.3.3.10 0.018
-
 acetyl-CoA pH 7.5, 30°C, mutant enzyme S89A Brassica juncea
2.3.3.10 0.024
-
 acetyl-CoA pH 7.5, 30°C, mutant enzyme S359A Brassica juncea
2.3.3.10 0.026
-
 acetyl-CoA pH 7.5, 30°C, mutant enzyme S356A Brassica juncea
2.3.3.10 0.027
-
 acetyl-CoA pH 7.5, 30°C, mutant enzyme N115A Brassica juncea
2.3.3.10 0.029
-
 acetyl-CoA pH 7.5, 30°C, mutant enzyme C212S Brassica juncea
2.3.3.10 0.043
-
 acetyl-CoA pH 7.5, 30°C, His6-BjHMGS1 wild-type enzyme Brassica juncea
2.3.3.10 0.052
-
 acetyl-CoA pH 7.5, 30°C, mutant enzyme R157A Brassica juncea
2.3.3.10 0.055
-
 acetyl-CoA pH 7.5, 30°C, mutant enzyme H188N Brassica juncea
2.3.3.10 0.12
-
 acetyl-CoA pH 7.5, 30°C, mutant enzyme H188N/S359A Brassica juncea

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
2.3.3.10 additional information no stimulation by Mg2+, Mn2+, Zn2+ and Co2+ Brassica juncea

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.3.3.10 acetyl-CoA + acetoacetyl-CoA + H2O Brassica juncea second enzyme in the cytoplasmic mevalonate pathway of isoprenoid biosynthesis (S)-3-hydroxy-3-methylglutaryl-CoA + CoA
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
2.3.3.10 Brassica juncea
-
-
-

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
2.3.3.10 0.1
-
-
 Brassica juncea

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.3.3.10 acetyl-CoA + acetoacetyl-CoA + H2O
-
 Brassica juncea (S)-3-hydroxy-3-methylglutaryl-CoA + CoA
-
 ?
2.3.3.10 acetyl-CoA + acetoacetyl-CoA + H2O second enzyme in the cytoplasmic mevalonate pathway of isoprenoid biosynthesis Brassica juncea (S)-3-hydroxy-3-methylglutaryl-CoA + CoA
-
 ?

Subunits

EC Number Subunits Comment Organism
2.3.3.10 dimer
-
 Brassica juncea

Synonyms

EC Number Synonyms Comment Organism
2.3.3.10 3-hydroxy-3-methylglutaryl-CoA synthase 1
-
 Brassica juncea
2.3.3.10 HMG-CoA synthase 1
-
 Brassica juncea

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.3.3.10 35
-
-
 Brassica juncea

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
2.3.3.10 30 45 about 70% of maximal activity at 30°C and at 45°C Brassica juncea

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.3.3.10 0.0017
-
 acetyl-CoA pH 7.5, 30°C, mutant enzyme R157A Brassica juncea
2.3.3.10 0.0029
-
 acetyl-CoA pH 7.5, 30°C, mutant enzyme H188N Brassica juncea
2.3.3.10 0.0041
-
 acetyl-CoA pH 7.5, 30°C, mutant enzyme C212S Brassica juncea
2.3.3.10 0.0076
-
 acetyl-CoA pH 7.5, 30°C, mutant enzyme S89A Brassica juncea
2.3.3.10 0.0161
-
 acetyl-CoA pH 7.5, 30°C, mutant enzyme S356A Brassica juncea
2.3.3.10 0.0234
-
 acetyl-CoA pH 7.5, 30°C, mutant enzyme N115A Brassica juncea
2.3.3.10 0.214
-
 acetyl-CoA pH 7.5, 30°C, mutant enzyme S359A Brassica juncea
2.3.3.10 0.415
-
 acetyl-CoA pH 7.5, 30°C, His6-BjHMGS1 wild-type enzyme Brassica juncea

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.3.3.10 8.5
-
-
 Brassica juncea

pH Range

EC Number pH Minimum pH Maximum Comment Organism
2.3.3.10 7.5 9.5 pH 7.5: about 60% of maximal activity, pH 9.5: about 30% of maximal activity Brassica juncea

Ki Value [mM]

EC Number Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
2.3.3.10 0.009
-
 (S)-3-hydroxy-3-methylglutaryl-CoA pH 7.5, 30°C, wild-type enzyme Brassica juncea
2.3.3.10 0.028
-
 acetoacetyl-CoA pH 7.5, 30°C, mutant enzyme S359A Brassica juncea
2.3.3.10 0.034
-
 (S)-3-hydroxy-3-methylglutaryl-CoA pH 7.5, 30°C, mutant enzyme H188N Brassica juncea
2.3.3.10 0.038
-
 acetoacetyl-CoA pH 7.5, 30°C, wild-type enzyme Brassica juncea
2.3.3.10 0.0389
-
 (S)-3-hydroxy-3-methylglutaryl-CoA pH 7.5, 30°C, mutant enzyme H188N/S359A Brassica juncea
2.3.3.10 0.039
-
 acetoacetyl-CoA pH 7.5, 30°C, mutant enzyme C212S Brassica juncea
2.3.3.10 0.046
-
 acetoacetyl-CoA pH 7.5, 30°C, mutant enzyme S89A Brassica juncea
2.3.3.10 0.08
-
 CoA pH 7.5, 30°C, wild-type enzyme Brassica juncea
2.3.3.10 0.091
-
 acetoacetyl-CoA pH 7.5, 30°C, mutant enzyme N115A Brassica juncea
2.3.3.10 0.199
-
 acetoacetyl-CoA pH 7.5, 30°C, mutant enzyme S356A Brassica juncea
2.3.3.10 0.242
-
 acetoacetyl-CoA pH 7.5, 30°C, mutant enzyme R157A Brassica juncea

IC50 Value

EC Number IC50 Value IC50 Value Maximum Comment Organism Inhibitor Structure
2.3.3.10 0.000035
-
 IC50: 35 nM Brassica juncea F-244