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Literature summary for 1.14.11.21 extracted from

  • Janc, J.W.; Egan, L.A.; Townsend, C.A.
    Purification and characterization of clavaminate synthase from Streptomyces antibioticus. A multifunctional enzyme of clavam biosynthesis (1995), J. Biol. Chem., 270, 5399-5404.
    View publication on PubMed
Search for more literature for 1.14.11.21

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0032
-
 Fe2+ pH 7.0, 22°C Streptomyces antibioticus
0.039
-
 2-oxoglutarate pH 7.0, 22°C Streptomyces antibioticus
0.38
-
 (2S,3R)-proclavaminate pH 7.0, 22°C Streptomyces antibioticus

Metals/Ions

Metals/Ions Comment Organism Structure
Fe2+ required Streptomyces antibioticus
Fe2+ required Streptomyces clavuligerus

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
35360
-
 CS1, mass spectrometry Streptomyces clavuligerus
35750
-
 CS2, mass spectrometry Streptomyces clavuligerus
35840
-
 CS3, mass spectrometry Streptomyces antibioticus
38000
-
 CS1, gel filtration Streptomyces clavuligerus
41000
-
 CS3, gel filtration Streptomyces antibioticus
42000
-
 CS2, gel filtration Streptomyces clavuligerus

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
deoxyamidinoproclavaminate + 2-oxoglutarate + O2 Streptomyces antibioticus
-
 amidinoproclavaminate + succinate + CO2 + H2O
-
 ?
deoxyamidinoproclavaminate + 2-oxoglutarate + O2 Streptomyces clavuligerus
-
 amidinoproclavaminate + succinate + CO2 + H2O
-
 ?
deoxyguanidinoproclavaminate + 2-oxoglutarate + O2 Streptomyces antibioticus
-
 guanidinoproclavaminate + succinate + CO2 + H2O
-
 ?
deoxyguanidinoproclavaminate + 2-oxoglutarate + O2 Streptomyces clavuligerus
-
 guanidinoproclavaminate + succinate + CO2 + H2O
-
 ?
deoxyguanidinoproclavaminate + 2-oxoglutarate + O2 Streptomyces antibioticus Tu1718
-
 guanidinoproclavaminate + succinate + CO2 + H2O
-
 ?
dihydroclavaminate + 2-oxoglutarate + O2 Streptomyces antibioticus
-
 clavaminate + succinate + CO2 + H2O
-
 ?
dihydroclavaminate + 2-oxoglutarate + O2 Streptomyces clavuligerus
-
 clavaminate + succinate + CO2 + H2O
-
 ?
additional information Streptomyces antibioticus enzyme plays a critical role in the biosynthesis of the clavam metabolites ?
-
 ?
additional information Streptomyces clavuligerus enzyme plays a critical role in the biosynthesis of the clavam metabolites ?
-
 ?
additional information Streptomyces antibioticus biosynthesic pathway of clavulanic acid, a beta-lactamase inhibitor ?
-
 ?
additional information Streptomyces clavuligerus biosynthesic pathway of clavulanic acid, a beta-lactamase inhibitor ?
-
 ?
additional information Streptomyces antibioticus Tu1718 enzyme plays a critical role in the biosynthesis of the clavam metabolites ?
-
 ?
additional information Streptomyces antibioticus Tu1718 biosynthesic pathway of clavulanic acid, a beta-lactamase inhibitor ?
-
 ?
proclavaminate + 2-oxoglutarate + O2 Streptomyces antibioticus
-
 dihydroclavaminate + succinate + CO2 + H2O
-
 ?
proclavaminate + 2-oxoglutarate + O2 Streptomyces clavuligerus
-
 dihydroclavaminate + succinate + CO2 + H2O
-
 ?

Organism

Organism UniProt Comment Textmining
Streptomyces antibioticus
-
 strain Tü 1718, CS3, no production of clavulanic acid
-
Streptomyces antibioticus Tu1718
-
 strain Tü 1718, CS3, no production of clavulanic acid
-
Streptomyces clavuligerus
-
 2 isozymes CS1 and CS2
-

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: 52 s, catalase partially protects Streptomyces clavuligerus

Purification (Commentary)

Purification (Comment) Organism
12.5fold to homogeneity Streptomyces antibioticus

Reaction

Reaction Comment Organism Reaction ID
deoxyamidinoproclavaminate + 2-oxoglutarate + O2 = amidinoproclavaminate + succinate + CO2 reaction mechanism Streptomyces antibioticus
a
deoxyamidinoproclavaminate + 2-oxoglutarate + O2 = amidinoproclavaminate + succinate + CO2 reaction mechanism Streptomyces clavuligerus
a
dihydroclavaminate + 2-oxoglutarate + O2 = clavaminate + succinate + CO2 + H2O reaction mechanism Streptomyces antibioticus
a
dihydroclavaminate + 2-oxoglutarate + O2 = clavaminate + succinate + CO2 + H2O reaction mechanism Streptomyces clavuligerus
a
proclavaminate + 2-oxoglutarate + O2 = dihydroclavaminate + succinate + CO2 + H2O reaction mechanism Streptomyces antibioticus
a
proclavaminate + 2-oxoglutarate + O2 = dihydroclavaminate + succinate + CO2 + H2O reaction mechanism Streptomyces clavuligerus
a

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.1
-
 purified enzyme Streptomyces antibioticus

Storage Stability

Storage Stability Organism
-80°C, purified protein, 50 mM MOPS, pH 7.0, 0.02 mM EDTA, 2 mM DTT, 50% glycerol, stable for at least 1 year Streptomyces antibioticus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
(2S,3R)-proclavaminate + 2-oxoglutarate + O2
-
 Streptomyces antibioticus 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 antibioticus ? + H2O2
-
 ?
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
-
 ?
deoxyamidinoproclavaminate + 2-oxoglutarate + O2
-
 Streptomyces antibioticus amidinoproclavaminate + succinate + CO2 + H2O
-
 ?
deoxyamidinoproclavaminate + 2-oxoglutarate + O2
-
 Streptomyces clavuligerus amidinoproclavaminate + succinate + CO2 + H2O
-
 ?
deoxyguanidinoproclavaminate + 2-oxoglutarate + O2
-
 Streptomyces antibioticus guanidinoproclavaminate + succinate + CO2 + H2O
-
 ?
deoxyguanidinoproclavaminate + 2-oxoglutarate + O2
-
 Streptomyces clavuligerus guanidinoproclavaminate + succinate + CO2 + H2O
-
 ?
deoxyguanidinoproclavaminate + 2-oxoglutarate + O2 hydroxylation Streptomyces antibioticus guanidinoproclavaminate + succinate + CO2 + H2O
-
 ?
deoxyguanidinoproclavaminate + 2-oxoglutarate + O2 hydroxylation Streptomyces clavuligerus guanidinoproclavaminate + succinate + CO2 + H2O
-
 ?
deoxyguanidinoproclavaminate + 2-oxoglutarate + O2
-
 Streptomyces antibioticus Tu1718 guanidinoproclavaminate + succinate + CO2 + H2O
-
 ?
deoxyguanidinoproclavaminate + 2-oxoglutarate + O2 hydroxylation Streptomyces antibioticus Tu1718 guanidinoproclavaminate + succinate + CO2 + H2O
-
 ?
dihydroclavaminate + 2-oxoglutarate + O2
-
 Streptomyces antibioticus clavaminate + succinate + CO2 + H2O
-
 ?
dihydroclavaminate + 2-oxoglutarate + O2
-
 Streptomyces clavuligerus clavaminate + succinate + CO2 + H2O
-
 ?
dihydroclavaminate + 2-oxoglutarate + O2 cyclization Streptomyces antibioticus clavaminate + succinate + CO2 + H2O
-
 ?
dihydroclavaminate + 2-oxoglutarate + O2 cyclization Streptomyces clavuligerus clavaminate + succinate + CO2 + H2O
-
 ?
additional information dependent on O2 and 2-oxoglutarate Streptomyces antibioticus ?
-
 ?
additional information dependent on O2 and 2-oxoglutarate Streptomyces clavuligerus ?
-
 ?
additional information enzyme plays a critical role in the biosynthesis of the clavam metabolites Streptomyces antibioticus ?
-
 ?
additional information enzyme plays a critical role in the biosynthesis of the clavam metabolites Streptomyces clavuligerus ?
-
 ?
additional information biosynthesic pathway of clavulanic acid, a beta-lactamase inhibitor Streptomyces antibioticus ?
-
 ?
additional information biosynthesic pathway of clavulanic acid, a beta-lactamase inhibitor Streptomyces clavuligerus ?
-
 ?
additional information dependent on O2 and 2-oxoglutarate Streptomyces antibioticus Tu1718 ?
-
 ?
additional information enzyme plays a critical role in the biosynthesis of the clavam metabolites Streptomyces antibioticus Tu1718 ?
-
 ?
additional information biosynthesic pathway of clavulanic acid, a beta-lactamase inhibitor Streptomyces antibioticus Tu1718 ?
-
 ?
proclavaminate + 2-oxoglutarate + O2
-
 Streptomyces antibioticus dihydroclavaminate + succinate + CO2 + H2O
-
 ?
proclavaminate + 2-oxoglutarate + O2
-
 Streptomyces clavuligerus dihydroclavaminate + succinate + CO2 + H2O
-
 ?
proclavaminate + 2-oxoglutarate + O2 saturation Streptomyces antibioticus 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 antibioticus 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 antibioticus 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 * 35357-45000, CS1, mass spectrometry and SDS-PAGE Streptomyces clavuligerus
monomer 1 * 35839-38000, CS3, mass spectrometry and SDS-PAGE Streptomyces antibioticus
monomer 1 * 35750-44000, CS2, mass spectrometry and SDS-PAGE Streptomyces clavuligerus

Synonyms

Synonyms Comment Organism
CS
-
 Streptomyces antibioticus
CS
-
 Streptomyces clavuligerus

pH Optimum

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