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Literature summary for 1.14.20.1 extracted from

  • Lee, H.J.; Lloyd, M.D.; Harlos, K.; Clifton, I.J.; Baldwin, J.E.; Schofield, C.J.
    Kinetic and crystallographic studies on deacetoxycephalosporin C synthase (DAOCS) (2001), J. Mol. Biol., 308, 937-948.
    View publication on PubMed

Crystallization (Commentary)

Crystallization (Comment) Organism
crystallization of wild-type enzyme and of the DELTAR306 mutant complexed with iron(II) and 2-oxoglutarate to 2.1 A and the DELTAR306A mutant complexed with iron(II), succinate and unhydrated carbon dioxide to 1.96 A Streptomyces clavuligerus

Protein Variants

Protein Variants Comment Organism
DELTAI305-310 mutant truncated by six residues, reduced turnover number for penicillin N compared to wild-type enzyme, lower KM-value for penicillin N compared to wild-type enzyme Streptomyces clavuligerus
DELTAK310-314 mutant truncated by five residues, increased turnover number for penicillin N compared to wild-type enzyme, lower KM-value for penicillin N compared to wild-type enzyme, lower Km-value for penicillin G compared to wild-type enzyme, slightly higher turnover number for penicillin G compared to wild-type enzyme Streptomyces clavuligerus
DELTAR306-310 mutant truncated by five residues, reduced turnover number for penicillin N compared to wild-type enzyme, lower KM-value for penicillin N compared to wild-type enzyme Streptomyces clavuligerus
DELTAR307-310 mutant in which residues 307-310 are excised, reduced turnover number for penicillin N compared to wild-type enzyme, higher KM-value for penicillin N compared to wild-type enzyme, increased KM-value for penicillin G compared to wild-type enzyme, increased turnover-number for penicillin G compared to wild-type enzyme Streptomyces clavuligerus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.025
-
Penicillin N mutant enzyme DELTAK310-314 Streptomyces clavuligerus
0.026
-
Penicillin N mutant enzyme DELTAR306-310 Streptomyces clavuligerus
0.034
-
Penicillin N mutant enzyme DELTAI305-310 Streptomyces clavuligerus
0.036
-
Penicillin N wild-type enzyme Streptomyces clavuligerus
0.044
-
Penicillin N mutant enzyme DELTAR307-310 Streptomyces clavuligerus
0.5
-
penicillin G mutant enzyme DELTAK310-314 Streptomyces clavuligerus
0.7
-
penicillin G wild-type enzyme Streptomyces clavuligerus
1.6
-
penicillin G mutant enzyme DELTAR307-310 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 the enzyme catalyzes the committed step in the biosynthesis of cephalosporin antibiotics deacetoxycephalosporin C + succinate + CO2 + H2O
-
?

Organism

Organism UniProt Comment Textmining
Streptomyces clavuligerus P18548
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant enzyme expressed in Escherichia coli Streptomyces clavuligerus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
penicillin G + 2-oxo-4-methylpentanoate + O2
-
Streptomyces clavuligerus phenylacetyl-7-aminodeacetoxy cephalosporanic acid + ?
-
?
penicillin G + 2-oxoglutarate + O2
-
Streptomyces clavuligerus phenylacetyl-7-aminodeacetoxy cephalosporanic acid + succinate + CO2 + H2O
-
?
penicillin G + 2-oxohexanoate + O2
-
Streptomyces clavuligerus phenylacetyl-7-aminodeacetoxy cephalosporanic acid + ?
-
?
penicillin N + 2-oxoglutarate + O2
-
Streptomyces clavuligerus deacetoxycephalosporin C + succinate + CO2 + H2O
-
?
penicillin N + 2-oxoglutarate + O2 the enzyme catalyzes the committed step in the biosynthesis of cephalosporin antibiotics Streptomyces clavuligerus deacetoxycephalosporin C + succinate + CO2 + H2O
-
?

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.02
-
Penicillin N mutant enzyme DELTAI305-310, DELTAR306-310 and mutant DELTAR307-310 Streptomyces clavuligerus
0.05
-
penicillin G wild-type enzyme Streptomyces clavuligerus
0.05
-
Penicillin N wild-type enzyme Streptomyces clavuligerus
0.06
-
penicillin G mutant enzyme DELTAK310-314 Streptomyces clavuligerus
0.08
-
penicillin G mutant enzyme DELTAR307-310 Streptomyces clavuligerus
0.08
-
Penicillin N mutant enzyme DELTAI310-314 Streptomyces clavuligerus