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Literature summary for 1.14.11.21 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

Inhibitors Comment Organism Structure
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]

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

Metals/Ions

Metals/Ions Comment Organism Structure
Fe2+ required Streptomyces clavuligerus

Molecular Weight [Da]

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

Natural Substrates/ Products (Substrates)

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

Organism

Organism UniProt Comment Textmining
Streptomyces clavuligerus
-
strain ATCC 27064
-

Oxidation Stability

Oxidation Stability Organism
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)

Purification (Comment) Organism
92fold Streptomyces clavuligerus

Reaction

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

Specific Activity [micromol/min/mg]

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

Storage Stability

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

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
(2S,3R)-proclavaminate + 2-oxoglutarate + O2
-
Streptomyces clavuligerus clavaminate + succinate + CO2 + H2O
-
r
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
-
?
dihydroclavaminate + 2-oxoglutarate + O2
-
Streptomyces clavuligerus clavaminate + succinate + CO2 + H2O
-
?
dihydroclavaminate + 2-oxoglutarate + O2 cyclization Streptomyces clavuligerus clavaminate + succinate + CO2 + H2O
-
?
additional information specific for 2-oxoglutarate Streptomyces clavuligerus ?
-
?
additional information stereospecificity Streptomyces clavuligerus ?
-
?
additional information biosynthesic pathway of clavulanic acid, a beta-lactamase inhibitor Streptomyces clavuligerus ?
-
?
proclavaminate + 2-oxoglutarate + O2
-
Streptomyces clavuligerus dihydroclavaminate + succinate + CO2 + H2O
-
?
proclavaminate + 2-oxoglutarate + O2 saturation Streptomyces clavuligerus dihydroclavaminate + succinate + CO2 + H2O
-
?
proclavaminate + 2-oxoglutarate + O2 no activity with the racemic erythro isomers (2S,3S) and (2R,3R) Streptomyces clavuligerus dihydroclavaminate + succinate + CO2 + H2O
-
?
proclavaminate + 2-oxoglutarate + O2 specific for the threo-(2S,3R) enantiomer Streptomyces clavuligerus dihydroclavaminate + succinate + CO2 + H2O
-
?

Subunits

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

Turnover Number [1/s]

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

pH Optimum

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