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Literature summary extracted from

  • Salowe, S.P.; Marsh, E.N.; Townsend, C.A.
    Purification and characterization of clavaminate synthase from Streptomyces clavuligerus: an unusual oxidative enzyme in natural product biosynthesis (1990), Biochemistry, 29, 6499-6508.
    View publication on PubMed

Inhibitors

EC Number Inhibitors Comment Organism Structure
1.14.11.21 ascorbate inactivation due to release of peroxide from reaction of ascorbate and O2 in absence of proclavaminate, but in presence of Fe2+, t1/2: 50 min, catalase protects Streptomyces clavuligerus

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.14.11.21 additional information
-
 additional information steady-state kinetics Streptomyces clavuligerus
1.14.11.21 0.0019
-
 Fe2+ pH 7.0, 22°C Streptomyces clavuligerus
1.14.11.21 0.042
-
 2-oxoglutarate pH 7.0, 22°C Streptomyces clavuligerus
1.14.11.21 0.19
-
 (2S,3R)-proclavaminate pH 7.0, 22°C Streptomyces clavuligerus
1.14.11.21 0.38
-
 rac-proclavaminate pH 7.0, 22°C Streptomyces clavuligerus

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
1.14.11.21 Fe2+ required Streptomyces clavuligerus

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.14.11.21 38000
-
 CS1, gel filtration Streptomyces clavuligerus

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.14.11.21 dihydroclavaminate + 2-oxoglutarate + O2 Streptomyces clavuligerus
-
 clavaminate + succinate + CO2 + H2O
-
 ?
1.14.11.21 additional information Streptomyces clavuligerus biosynthesic pathway of clavulanic acid, a beta-lactamase inhibitor ?
-
 ?
1.14.11.21 proclavaminate + 2-oxoglutarate + O2 Streptomyces clavuligerus
-
 dihydroclavaminate + succinate + CO2 + H2O
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
1.14.11.21 Streptomyces clavuligerus
-
 strain ATCC 27064
-

Oxidation Stability

EC Number Oxidation Stability Organism
1.14.11.21 inactivation due to release of peroxide from reaction of 2-oxoglutarate and O2 in absence of proclavaminate, in presence of Fe2+, t1/2: 5 min, with ascorbate instead of 2-oxoglutarate t1/2: 50 min, catalase protects Streptomyces clavuligerus

Purification (Commentary)

EC Number Purification (Comment) Organism
1.14.11.21 92fold Streptomyces clavuligerus

Reaction

EC Number Reaction Comment Organism Reaction ID
1.14.11.21 dihydroclavaminate + 2-oxoglutarate + O2 = clavaminate + succinate + CO2 + H2O kinetic mechanism Streptomyces clavuligerus
a
1.14.11.21 proclavaminate + 2-oxoglutarate + O2 = dihydroclavaminate + succinate + CO2 + H2O kinetic mechanism Streptomyces clavuligerus
a

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.14.11.21 0.57
-
 purified enzyme, 40°C Streptomyces clavuligerus

Storage Stability

EC Number Storage Stability Organism
1.14.11.21 -70°C, purified enzyme, frozen in liquid N2, stable for at least several months Streptomyces clavuligerus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.14.11.21 (2S,3R)-proclavaminate + 2-oxoglutarate + O2
-
 Streptomyces clavuligerus clavaminate + succinate + CO2 + H2O
-
 r
1.14.11.21 2-oxoglutarate + O2 side reaction in absence of proclavaminate, oxidative decarboxylation of 2-oxoglutarate in presence of Fe2+ and formation of peroxide, which inactivates the enzyme Streptomyces clavuligerus ? + H2O2
-
 ?
1.14.11.21 dihydroclavaminate + 2-oxoglutarate + O2
-
 Streptomyces clavuligerus clavaminate + succinate + CO2 + H2O
-
 ?
1.14.11.21 dihydroclavaminate + 2-oxoglutarate + O2 cyclization Streptomyces clavuligerus clavaminate + succinate + CO2 + H2O
-
 ?
1.14.11.21 additional information specific for 2-oxoglutarate Streptomyces clavuligerus ?
-
 ?
1.14.11.21 additional information stereospecificity Streptomyces clavuligerus ?
-
 ?
1.14.11.21 additional information biosynthesic pathway of clavulanic acid, a beta-lactamase inhibitor Streptomyces clavuligerus ?
-
 ?
1.14.11.21 proclavaminate + 2-oxoglutarate + O2
-
 Streptomyces clavuligerus dihydroclavaminate + succinate + CO2 + H2O
-
 ?
1.14.11.21 proclavaminate + 2-oxoglutarate + O2 saturation Streptomyces clavuligerus dihydroclavaminate + succinate + CO2 + H2O
-
 ?
1.14.11.21 proclavaminate + 2-oxoglutarate + O2 no activity with the racemic erythro isomers (2S,3S) and (2R,3R) Streptomyces clavuligerus dihydroclavaminate + succinate + CO2 + H2O
-
 ?
1.14.11.21 proclavaminate + 2-oxoglutarate + O2 specific for the threo-(2S,3R) enantiomer Streptomyces clavuligerus dihydroclavaminate + succinate + CO2 + H2O
-
 ?

Subunits

EC Number Subunits Comment Organism
1.14.11.21 monomer 1* 47000, enzyme form 1, SDS-PAGE Streptomyces clavuligerus
1.14.11.21 monomer 1* 46000, enzyme form 2, SDS-PAGE Streptomyces clavuligerus

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.14.11.21 0.725
-
 rac-proclavaminate pH 7.0, 40°C Streptomyces clavuligerus
1.14.11.21 0.765
-
 (2S,3R)-proclavaminate pH 7.0, 40°C Streptomyces clavuligerus

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.14.11.21 7
-
 assay at Streptomyces clavuligerus