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Literature summary extracted from

  • Khaleeli, N.; Busby, R.W.; Townsend, C.A.
    Site-directed mutagenesis and biochemical analysis of the endogenous ligands in the ferrous active site of clavaminate synthase. The His-3 variant of the 2-His-1-carboxylate model (2000), Biochemistry, 39, 8666-8673.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.14.11.21 gene cs2, isozyme CS2, wild-type and mutants, overexpression in Escherichia coli JM101 Streptomyces clavuligerus

Protein Variants

EC Number Protein Variants Comment Organism
1.14.11.21 H109Q isozyme CS2, site-directed mutagensis, 83% activity of clavaminate formation compared to the wild-type enzyme, unaltered hydroxylating activity Streptomyces clavuligerus
1.14.11.21 H122Q isozyme CS2, site-directed mutagensis, 96% activity of clavaminate formation compared to the wild-type enzyme, unaltered hydroxylating activity Streptomyces clavuligerus
1.14.11.21 H131Q isozyme CS2, site-directed mutagensis, 16% activity of clavaminate formation compared to the wild-type enzyme, unaltered hydroxylating activity Streptomyces clavuligerus
1.14.11.21 H145Q isozyme CS2, site-directed mutagensis, no activity of clavaminate formation, but slight hydroxylating activity of deoxyguanidinoproclavaminate Streptomyces clavuligerus
1.14.11.21 H151Q isozyme CS2, site-directed mutagensis, 10% activity of clavaminate formation compared to the wild-type enzyme, unaltered hydroxylating activity Streptomyces clavuligerus
1.14.11.21 H167Q isozyme CS2, site-directed mutagensis, 95% activity of clavaminate formation compared to the wild-type enzyme, unaltered hydroxylating activity Streptomyces clavuligerus
1.14.11.21 H280Q isozyme CS2, site-directed mutagensis, no activity of clavaminate formation, but slight hydroxylating activity of deoxyguanidinoproclavaminate Streptomyces clavuligerus
1.14.11.21 H297Q isozyme CS2, site-directed mutagensis, 65% activity of clavaminate formation compared to the wild-type enzyme, unaltered hydroxylating activity Streptomyces clavuligerus

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
1.14.11.21 Fe2+ required Streptomyces clavuligerus
1.14.11.21 Fe2+ isozyme CS2, a single ferrous active site Streptomyces clavuligerus

Organism

EC Number Organism UniProt Comment Textmining
1.14.11.21 Streptomyces clavuligerus
-
isozyme CS2
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.14.11.21 recombinant wild-type and mutants of isozyme CS2 from Escherichia coli JM101 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 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 proclavaminate + 2-oxoglutarate + O2 saturation Streptomyces clavuligerus dihydroclavaminate + succinate + CO2 + H2O
-
?

Synonyms

EC Number Synonyms Comment Organism
1.14.11.21 CS
-
Streptomyces clavuligerus

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.14.11.21 23
-
assay at Streptomyces clavuligerus

pH Optimum

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