Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary for 1.14.20.1 extracted from

  • Oester, L.M.; van Scheltinga, A.C.; Valegard, K.; Hose, A.M.; Dubus, A.; Hajdu, J.; Andersson, I.
    Conformational flexibility of the C terminus with implications for substrate binding and catalysis revealed in a new crystal form of deacetoxycephalosporin C synthase (2004), J. Mol. Biol., 343, 157-171.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expression of wild-type and C-terminally truncated mutant enzymes as N-terminally His-tagged proteins in Escherichia coli strain BL21(DE3) Streptomyces clavuligerus

Crystallization (Commentary)

Crystallization (Comment) Organism
recombinant N-terminally His-tagged wild-type and C-terminally truncated mutant enzymes, free enzyme, or complexing with Fe2+, or Fe2+/ampicillin, hanging drop vapour diffusion method, 4°C, precipitant solution: 100 mM HEPES-NaOH, pH 8.0, 0.9-1.1 M ammonium sulfate, the reservoir solution is covered with oil to retard the evaporation, cryoprotection by 30% v/v ethylene glycol in precipitant solution, X-ray structure determination and analysis at 2.3 A resolution, molecular modeling Streptomyces clavuligerus

Protein Variants

Protein Variants Comment Organism
additional information shortening of the C-terminus by more than 4 residues reduces enzyme activity and alters the crystal structure from merohedrally twinned trimeric crystals to a non-trimeric structure with a free C-terminal arm, crystals show no twinning Streptomyces clavuligerus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.89
-
penicillin G wild-type enzyme, pH 7.5, 30°C Streptomyces clavuligerus
20.6
-
penicillin G recombinant His-tagged enzyme, pH 7.5, 30°C Streptomyces clavuligerus

Localization

Localization Comment Organism GeneOntology No. Textmining

Metals/Ions

Metals/Ions Comment Organism Structure
Fe2+ bound to the active site, forms a reactive iron-oxygen reaction intermediate during catalysis, complex structure in native and His-tagged enzyme Streptomyces clavuligerus

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
additional information
-
gel filtration Streptomyces clavuligerus

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
penicillin N + 2-oxoglutarate + O2 Streptomyces clavuligerus
-
deacetoxycephalosporin C + succinate + CO2 + H2O
-
ir

Organism

Organism UniProt Comment Textmining
Streptomyces clavuligerus P18548
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant N-terminally His-tagged wild-type and C-terminally truncated mutant enzymes from Escherichia coli strain BL21(DE3) Streptomyces clavuligerus

Reaction

Reaction Comment Organism Reaction ID
penicillin N + 2-oxoglutarate + O2 = deacetoxycephalosporin C + succinate + CO2 + H2O reaction mechanism Streptomyces clavuligerus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
penicillin G + 2-oxoglutarate + O2
-
Streptomyces clavuligerus phenylacetyl-7-aminodeacetoxycephalosporanic acid + succinate + CO2 + H2O
-
ir
penicillin N + 2-oxoglutarate + O2
-
Streptomyces clavuligerus deacetoxycephalosporin C + succinate + CO2 + H2O
-
ir
penicillin N + 2-oxoglutarate + O2 oxidative ring expansion via reactive iron-oxygen intermediate Streptomyces clavuligerus deacetoxycephalosporin C + succinate + CO2 + H2O
-
ir

Subunits

Subunits Comment Organism
monomer His-tagged enzyme in solution Streptomyces clavuligerus
More active site structure in the apo-enzyme Streptomyces clavuligerus
trimer wild-type enzyme in crystals Streptomyces clavuligerus

Synonyms

Synonyms Comment Organism
DAOCS
-
Streptomyces clavuligerus
deacetoxycephalosporin C synthase
-
Streptomyces clavuligerus

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
30
-
assay at Streptomyces clavuligerus

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.056
-
penicillin G recombinant His-tagged enzyme, pH 7.5, 30°C Streptomyces clavuligerus
0.079
-
penicillin G wild-type enzyme, pH 7.5, 30°C Streptomyces clavuligerus

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
assay at Streptomyces clavuligerus