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Literature summary extracted from

  • Busby, R.W.; Chang, M.D.T.; Busby, R.C.; Wimp, J.; Townsend, C.A.
    Expression and purification of two isoenzymes of clavaminate synthase and initial characterization of the iron binding site. General error analysis in polymerase chain reaction amplification (1995), J. Biol. Chem., 270, 4262-4269.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.14.11.21 isozymes CS1 and CS2, overexpression in Escherichia coli JM101 Streptomyces clavuligerus

General Stability

EC Number General Stability Organism
1.14.11.21 Co2+ completely inhibits the enzyme, competitive to Fe2+, but also stabilizes the enzyme against self-inactivation in absence of proclavaminate Streptomyces clavuligerus

Inhibitors

EC Number Inhibitors Comment Organism Structure
1.14.11.21 Co2+ complete inhibition competitive to Fe2+, but stabilizes the enzyme against self-inactivation in absence of proclavaminate Streptomyces clavuligerus
1.14.11.21 diethyl dicarbonate isozyme CS2, rapid inactivation at pH 6.0, 25°C, t1/2: 1.1 min Streptomyces clavuligerus
1.14.11.21 N-ethylmaleimide inactivation Streptomyces clavuligerus

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.14.11.21 0.13
-
proclavaminate isozyme CS2, pH 7.0, 22°C Streptomyces clavuligerus
1.14.11.21 0.22
-
proclavaminate isozyme CS1, 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 35360
-
recombinant enzyme Streptomyces clavuligerus
1.14.11.21 35360
-
CS1, mass spectrometry Streptomyces clavuligerus
1.14.11.21 35750
-
recombinant enzyme Streptomyces clavuligerus
1.14.11.21 35750
-
CS2, mass spectrometry 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 deoxyguanidinoproclavaminate + 2-oxoglutarate + O2 Streptomyces clavuligerus
-
guanidinoproclavaminate + succinate + CO2 + H2O
-
?

Organism

EC Number Organism UniProt Comment Textmining
1.14.11.21 Streptomyces clavuligerus
-
2 isozymes CS1 and CS2
-

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 recombinant isozymes CS1 and CS2 to homogeneity, 10fold Streptomyces clavuligerus

Reaction

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

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 0.8
-
purified recombinant isozyme CS2 Streptomyces clavuligerus
1.14.11.21 0.86
-
purified recombinant isozyme CS1 Streptomyces clavuligerus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.14.11.21 deoxyguanidinoproclavaminate + 2-oxoglutarate + O2
-
Streptomyces clavuligerus guanidinoproclavaminate + succinate + CO2 + H2O
-
?
1.14.11.21 deoxyguanidinoproclavaminate + 2-oxoglutarate + O2 hydroxylation Streptomyces clavuligerus guanidinoproclavaminate + succinate + CO2 + H2O
-
?
1.14.11.21 dihydroclavaminate + 2-oxoglutarate + O2 cyclization Streptomyces clavuligerus clavaminate + succinate + CO2 + H2O
-
?
1.14.11.21 additional information 1 or more histidine as well as cysteine residue located at or near the active site or in the metal binding site, critical for catalysis Streptomyces clavuligerus ?
-
?
1.14.11.21 additional information dependent on O2 and 2-oxoglutarate Streptomyces clavuligerus ?
-
?
1.14.11.21 proclavaminate + 2-oxoglutarate + O2 saturation Streptomyces clavuligerus dihydroclavaminate + succinate + CO2 + H2O
-
?

Synonyms

EC Number Synonyms Comment Organism
1.14.11.21 CS
-
Streptomyces clavuligerus

pH Optimum

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