BRENDA - Enzyme Database show
show all sequences of 1.14.11.21

Purification and characterization of clavaminate synthase from Streptomyces clavuligerus: an unusual oxidative enzyme in natural product biosynthesis

Salowe, S.P.; Marsh, E.N.; Townsend, C.A.; Biochemistry 29, 6499-6508 (1990)

Data extracted from this reference:

Inhibitors
Inhibitors
Commentary
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
Commentary
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
Commentary
Organism
Structure
Fe2+
required
Streptomyces clavuligerus
Molecular Weight [Da]
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
38000
-
CS1, gel filtration
Streptomyces clavuligerus
Natural Substrates/ Products (Substrates)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
dihydroclavaminate + 2-oxoglutarate + O2
Streptomyces clavuligerus
-
clavaminate + succinate + CO2 + H2O
-
Streptomyces clavuligerus
?
additional information
Streptomyces clavuligerus
biosynthesic pathway of clavulanic acid, a beta-lactamase inhibitor
?
-
-
-
proclavaminate + 2-oxoglutarate + O2
Streptomyces clavuligerus
-
dihydroclavaminate + succinate + CO2 + H2O
-
Streptomyces clavuligerus
?
Organism
Organism
Primary Accession No. (UniProt)
Commentary
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)
Commentary
Organism
92fold
Streptomyces clavuligerus
Reaction
Reaction
Commentary
Organism
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]
Commentary
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
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
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
639270
Streptomyces clavuligerus
? + H2O2
-
-
-
?
dihydroclavaminate + 2-oxoglutarate + O2
-
639270
Streptomyces clavuligerus
clavaminate + succinate + CO2 + H2O
-
639270
Streptomyces clavuligerus
?
dihydroclavaminate + 2-oxoglutarate + O2
cyclization
639270
Streptomyces clavuligerus
clavaminate + succinate + CO2 + H2O
-
639270
Streptomyces clavuligerus
?
additional information
specific for 2-oxoglutarate
639270
Streptomyces clavuligerus
?
-
-
-
-
additional information
stereospecificity
639270
Streptomyces clavuligerus
?
-
-
-
-
additional information
biosynthesic pathway of clavulanic acid, a beta-lactamase inhibitor
639270
Streptomyces clavuligerus
?
-
-
-
-
proclavaminate + 2-oxoglutarate + O2
-
639270
Streptomyces clavuligerus
dihydroclavaminate + succinate + CO2 + H2O
-
639270
Streptomyces clavuligerus
?
proclavaminate + 2-oxoglutarate + O2
saturation
639270
Streptomyces clavuligerus
dihydroclavaminate + succinate + CO2 + H2O
-
639270
Streptomyces clavuligerus
?
proclavaminate + 2-oxoglutarate + O2
no activity with the racemic erythro isomers (2S,3S) and (2R,3R)
639270
Streptomyces clavuligerus
dihydroclavaminate + succinate + CO2 + H2O
-
639270
Streptomyces clavuligerus
?
proclavaminate + 2-oxoglutarate + O2
specific for the threo-(2S,3R) enantiomer
639270
Streptomyces clavuligerus
dihydroclavaminate + succinate + CO2 + H2O
-
639270
Streptomyces clavuligerus
?
Subunits
Subunits
Commentary
Organism
monomer
1* 46000, enzyme form 2, SDS-PAGE; 1* 47000, enzyme form 1, SDS-PAGE
Streptomyces clavuligerus
Turnover Number [1/s]
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
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
Commentary
Organism
7
-
assay at
Streptomyces clavuligerus
Inhibitors (protein specific)
Inhibitors
Commentary
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] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
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 (protein specific)
Metals/Ions
Commentary
Organism
Structure
Fe2+
required
Streptomyces clavuligerus
Molecular Weight [Da] (protein specific)
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
38000
-
CS1, gel filtration
Streptomyces clavuligerus
Natural Substrates/ Products (Substrates) (protein specific)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
dihydroclavaminate + 2-oxoglutarate + O2
Streptomyces clavuligerus
-
clavaminate + succinate + CO2 + H2O
-
Streptomyces clavuligerus
?
additional information
Streptomyces clavuligerus
biosynthesic pathway of clavulanic acid, a beta-lactamase inhibitor
?
-
-
-
proclavaminate + 2-oxoglutarate + O2
Streptomyces clavuligerus
-
dihydroclavaminate + succinate + CO2 + H2O
-
Streptomyces clavuligerus
?
Oxidation Stability (protein specific)
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) (protein specific)
Commentary
Organism
92fold
Streptomyces clavuligerus
Specific Activity [micromol/min/mg] (protein specific)
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
0.57
-
purified enzyme, 40°C
Streptomyces clavuligerus
Storage Stability (protein specific)
Storage Stability
Organism
-70°C, purified enzyme, frozen in liquid N2, stable for at least several months
Streptomyces clavuligerus
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
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
639270
Streptomyces clavuligerus
? + H2O2
-
-
-
?
dihydroclavaminate + 2-oxoglutarate + O2
-
639270
Streptomyces clavuligerus
clavaminate + succinate + CO2 + H2O
-
639270
Streptomyces clavuligerus
?
dihydroclavaminate + 2-oxoglutarate + O2
cyclization
639270
Streptomyces clavuligerus
clavaminate + succinate + CO2 + H2O
-
639270
Streptomyces clavuligerus
?
additional information
specific for 2-oxoglutarate
639270
Streptomyces clavuligerus
?
-
-
-
-
additional information
stereospecificity
639270
Streptomyces clavuligerus
?
-
-
-
-
additional information
biosynthesic pathway of clavulanic acid, a beta-lactamase inhibitor
639270
Streptomyces clavuligerus
?
-
-
-
-
proclavaminate + 2-oxoglutarate + O2
-
639270
Streptomyces clavuligerus
dihydroclavaminate + succinate + CO2 + H2O
-
639270
Streptomyces clavuligerus
?
proclavaminate + 2-oxoglutarate + O2
saturation
639270
Streptomyces clavuligerus
dihydroclavaminate + succinate + CO2 + H2O
-
639270
Streptomyces clavuligerus
?
proclavaminate + 2-oxoglutarate + O2
no activity with the racemic erythro isomers (2S,3S) and (2R,3R)
639270
Streptomyces clavuligerus
dihydroclavaminate + succinate + CO2 + H2O
-
639270
Streptomyces clavuligerus
?
proclavaminate + 2-oxoglutarate + O2
specific for the threo-(2S,3R) enantiomer
639270
Streptomyces clavuligerus
dihydroclavaminate + succinate + CO2 + H2O
-
639270
Streptomyces clavuligerus
?
Subunits (protein specific)
Subunits
Commentary
Organism
monomer
1* 46000, enzyme form 2, SDS-PAGE; 1* 47000, enzyme form 1, SDS-PAGE
Streptomyces clavuligerus
Turnover Number [1/s] (protein specific)
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
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 (protein specific)
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7
-
assay at
Streptomyces clavuligerus
Other publictions for EC 1.14.11.21
No.
1st author
Pub Med
title
organims
journal
volume
pages
year
Activating Compound
Application
Cloned(Commentary)
Crystallization (Commentary)
Engineering
General Stability
Inhibitors
KM Value [mM]
Localization
Metals/Ions
Molecular Weight [Da]
Natural Substrates/ Products (Substrates)
Organic Solvent Stability
Organism
Oxidation Stability
Posttranslational Modification
Purification (Commentary)
Reaction
Renatured (Commentary)
Source Tissue
Specific Activity [micromol/min/mg]
Storage Stability
Substrates and Products (Substrate)
Subunits
Temperature Optimum [°C]
Temperature Range [°C]
Temperature Stability [°C]
Turnover Number [1/s]
pH Optimum
pH Range
pH Stability
Cofactor
Ki Value [mM]
pI Value
IC50 Value
Activating Compound (protein specific)
Application (protein specific)
Cloned(Commentary) (protein specific)
Cofactor (protein specific)
Crystallization (Commentary) (protein specific)
Engineering (protein specific)
General Stability (protein specific)
IC50 Value (protein specific)
Inhibitors (protein specific)
Ki Value [mM] (protein specific)
KM Value [mM] (protein specific)
Localization (protein specific)
Metals/Ions (protein specific)
Molecular Weight [Da] (protein specific)
Natural Substrates/ Products (Substrates) (protein specific)
Organic Solvent Stability (protein specific)
Oxidation Stability (protein specific)
Posttranslational Modification (protein specific)
Purification (Commentary) (protein specific)
Renatured (Commentary) (protein specific)
Source Tissue (protein specific)
Specific Activity [micromol/min/mg] (protein specific)
Storage Stability (protein specific)
Substrates and Products (Substrate) (protein specific)
Subunits (protein specific)
Temperature Optimum [°C] (protein specific)
Temperature Range [°C] (protein specific)
Temperature Stability [°C] (protein specific)
Turnover Number [1/s] (protein specific)
pH Optimum (protein specific)
pH Range (protein specific)
pH Stability (protein specific)
pI Value (protein specific)
Expression
General Information
General Information (protein specific)
Expression (protein specific)
KCat/KM [mM/s]
KCat/KM [mM/s] (protein specific)
742159
Shrestha
-
Heterologous production of cl ...
Streptomyces clavuligerus
Biotechnol. Bioprocess Eng.
22
359-365
2017
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723902
Chin
-
Predicting the catalytic sites ...
Streptomyces clavuligerus
Afr. J. Microbiol. Res.
5
3357-3366
2011
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685142
Borowski
Mechanism for cyclization reac ...
Streptomyces clavuligerus
Biochemistry
46
3682-3691
2007
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639036
Townsend
New reactions in clavulanic ac ...
Streptomyces clavuligerus
Curr. Opin. Chem. Biol.
6
583-589
2002
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639286
Zhang
Crystal structure of a clavami ...
Streptomyces clavuligerus
FEBS Lett.
517
7-12
2002
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639284
Zhou
Spectroscopic studies of subst ...
Streptomyces clavuligerus
J. Am. Chem. Soc.
123
7388-7398
2001
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11
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639281
Zhang
Structural origins of the sele ...
Streptomyces clavuligerus
Nat. Struct. Biol.
7
127-133
2000
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639282
Khaleeli
Site-directed mutagenesis and ...
Streptomyces clavuligerus
Biochemistry
39
8666-8673
2000
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639283
Doan
Mutagenesis studies on the iro ...
Streptomyces clavuligerus
Biochem. Biophys. Res. Commun.
279
240-244
2000
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15
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639280
Lloyd
-
Product-substrate engineering ...
Streptomyces clavuligerus
Tetrahedron
55
10201-10220
1999
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639279
Zhou
-
Substrate binding to the alpha ...
Streptomyces clavuligerus
J. Am. Chem. Soc.
120
13539-13540
1998
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639278
Baldwin
-
Chemo-enzymic synthesis of bic ...
Streptomyces clavuligerus
Tetrahedron
53
7011-7020
1997
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639277
Busby
A single monomeric iron center ...
Streptomyces clavuligerus
Bioorg. Med. Chem.
4
1059-1064
1996
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11
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10
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