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Literature summary for 1.14.11.21 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)

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

General Stability

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

Inhibitors

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

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.13
-
proclavaminate isozyme CS2, pH 7.0, 22°C Streptomyces clavuligerus
0.22
-
proclavaminate isozyme CS1, 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
35360
-
recombinant enzyme Streptomyces clavuligerus
35360
-
CS1, mass spectrometry Streptomyces clavuligerus
35750
-
recombinant enzyme Streptomyces clavuligerus
35750
-
CS2, mass spectrometry Streptomyces clavuligerus

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
deoxyguanidinoproclavaminate + 2-oxoglutarate + O2 Streptomyces clavuligerus
-
guanidinoproclavaminate + succinate + CO2 + H2O
-
?

Organism

Organism UniProt Comment Textmining
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: 5 min, with ascorbate instead of 2-oxoglutarate t1/2: 50 min, catalase protects Streptomyces clavuligerus

Purification (Commentary)

Purification (Comment) Organism
recombinant isozymes CS1 and CS2 to homogeneity, 10fold Streptomyces clavuligerus

Reaction

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

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.8
-
purified recombinant isozyme CS2 Streptomyces clavuligerus
0.86
-
purified recombinant isozyme CS1 Streptomyces clavuligerus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
deoxyguanidinoproclavaminate + 2-oxoglutarate + O2
-
Streptomyces clavuligerus guanidinoproclavaminate + succinate + CO2 + H2O
-
?
deoxyguanidinoproclavaminate + 2-oxoglutarate + O2 hydroxylation Streptomyces clavuligerus guanidinoproclavaminate + succinate + CO2 + H2O
-
?
dihydroclavaminate + 2-oxoglutarate + O2 cyclization Streptomyces clavuligerus clavaminate + succinate + CO2 + H2O
-
?
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 ?
-
?
additional information dependent on O2 and 2-oxoglutarate Streptomyces clavuligerus ?
-
?
proclavaminate + 2-oxoglutarate + O2 saturation Streptomyces clavuligerus dihydroclavaminate + succinate + CO2 + H2O
-
?

Synonyms

Synonyms Comment Organism
CS
-
Streptomyces clavuligerus

pH Optimum

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