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

BRENDA support

Literature summary extracted from

  • Lloyd, M.D.; Lipscomb, S.J.; Hewitson, K.S.; Hensgens, C.M.H.; Baldwin, J.E.; Schofield, C.J.
    Controlling the substrate selectivity of deacetoxycephalosporin/deacetylcephalosporin C synthase (2004), J. Biol. Chem., 279, 15420-15426.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.14.20.1 expression of wild-type and mutant enzymes in Escherichia coli strain BL21(DE3), subcloning in strain XL-1 Blue Acremonium chrysogenum

Protein Variants

EC Number Protein Variants Comment Organism
1.14.11.26 M306I hydroxylation reaction of deacetoxycephalosporin C, EC 1.14.11.26, is abolished, 59% of wild-type ring expansion activity Acremonium chrysogenum
1.14.11.26 additional information truncation of C-terminus to residue 310, 2fold enhancement of ring expansion reaction of penicillin G. Double mutant with truncation at residue 310 and M306I, selective catalyzation of ring expansion. Triple mutant with truncation at residue 310, M306I and N305L, selective catalization of ring expansion with improved kinetic parameters Acremonium chrysogenum
1.14.11.26 N305L 107% of wild-type ring expansion activity, 85% of wild-type hydroxylation activity Acremonium chrysogenum
1.14.11.26 W82A 5.5% of wild-type ring expansion activity, 71% of wild-type hydroxylation activity Acremonium chrysogenum
1.14.11.26 W82A ring expansion reaction of EC 1.14.20.1 is reduced Acremonium chrysogenum
1.14.11.26 W82S 44% of wild-type ring expansion activity, 18% of wild-type hydroxylation activity Acremonium chrysogenum
1.14.20.1 DELTA310/M306I site-directed mutagenesis, mutant selectively catalyzed the ring expansion with kinetics similar to the wild-type enzyme Acremonium chrysogenum
1.14.20.1 DELTA310/N305L/M306I site-directed mutagenesis, mutant selectively catalyzed the ring expansion, and shows a lower Km value than the wild-type enzyme and about 3fold improved kinetic parameters Acremonium chrysogenum
1.14.20.1 M306I site-directed mutagenesis, mutant shows no hydroxylation of desacetylcephalosporin C Acremonium chrysogenum
1.14.20.1 M306I hydroxylation reaction of deacetoxycephalosporin C, E C1.14.11.26, is abolished, 59% of wild-type ring expansion activity Acremonium chrysogenum
1.14.20.1 additional information DELTA310 is a C-terminally truncated mutant lacking residues 1-309, the mutant shows 2fold enhanced ring expansion activity with the artificial substrate penicillin G Acremonium chrysogenum
1.14.20.1 additional information truncation of C-terminus to residue 310, 2fold enhancement of ring expansion reaction of penicillin G. Double mutant with truncation at residue 310 and M306I, selective catalyzation of ring expansion. Triple mutant with truncation at residue 310, M306I and N305L, selective catalization of ring expansion with improved kinetic parameters Acremonium chrysogenum
1.14.20.1 N305L 107% of wild-type ring expansion activity, 85% of wild-type hydroxylation activity Acremonium chrysogenum
1.14.20.1 W82A site-directed mutagenesis, mutant shows reduced ring expansion activity with artificial substrate penicillin G Acremonium chrysogenum
1.14.20.1 W82A 5.5% of wild-type ring expansion activity, 71% of wild-type hydroxylation activity Acremonium chrysogenum
1.14.20.1 W82A ring expansion reaction of EC 1.14.20.1 is reduced Acremonium chrysogenum
1.14.20.1 W82S 44% of wild-type ring expansion activity, 18% of wild-type hydroxylation activity Acremonium chrysogenum

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.14.20.1 2
-
 penicillin G pH 7.5, 30°C, recombinant mutant DELTA310/N305L/M306I Acremonium chrysogenum
1.14.20.1 2
-
 penicillin G 30°C, triple mutant with truncation at residue 310, M306I and N305L Acremonium chrysogenum
1.14.20.1 2.41
-
 penicillin G 30°C, mutant N305L Acremonium chrysogenum
1.14.20.1 4.79
-
 penicillin G 30°C, mutant M306I Acremonium chrysogenum
1.14.20.1 6.02
-
 penicillin G pH 7.5, 30°C, recombinant wild-type enzyme Acremonium chrysogenum
1.14.20.1 6.04
-
 penicillin G 30°C, wild-type Acremonium chrysogenum
1.14.20.1 6.42
-
 penicillin G 30°C, double mutant with truncation at residue 310 and M306I Acremonium chrysogenum

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
1.14.20.1 Fe2+ dependent on, iron cofactor is bound by D184, H186, and H244 Acremonium chrysogenum

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.14.20.1 36348
-
 x * 36348, wild-type enzyme, electrospray mass spectrometry Acremonium chrysogenum

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.14.20.1 deacetoxycephalosporin C + 2-oxoglutarate + O2 Acremonium chrysogenum
-
 deacetylcephalosporin C + succinate + CO2
-
 ir
1.14.20.1 penicillin N + 2-oxoglutarate + O2 Acremonium chrysogenum
-
 deacetoxycephalosporin C + succinate + CO2 + H2O
-
 ir

Organism

EC Number Organism UniProt Comment Textmining
1.14.11.26 Acremonium chrysogenum
-
 catalyzes both reaction of EC 1.14.20.1 and of EC 1.14.11.26
-
1.14.20.1 Acremonium chrysogenum
-
 catalyzes both reaction of EC 1.14.20.1 and of EC 1.14.11.26
-
1.14.20.1 Acremonium chrysogenum P11935 bifunctional enzyme
-

Reaction

EC Number Reaction Comment Organism Reaction ID
1.14.20.1 penicillin N + 2-oxoglutarate + O2 = deacetoxycephalosporin C + succinate + CO2 + H2O bifunctional enzyme, residues W82, N305 and M306 are involved in catalysis, overview Acremonium chrysogenum
a

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.14.20.1 additional information
-
 activities of wild-type enzyme in ring-expansion and hydroxylation, cosubstrate and substrate conversion rate Acremonium chrysogenum
1.14.20.1 0.156
-
 purified recombinant wild-type enzyme, substrate 6-alpha-methylpenicillin N, substrate conversion Acremonium chrysogenum

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.14.11.26 7-aminodeacetoxycephalosporanic acid + 2-oxoglutarate + O2
-
 Acremonium chrysogenum 7-aminodeacetylcephalosporin + succinate + CO2
-
 ?
1.14.11.26 cephalexin + 2-oxoglutarate + O2
-
 Acremonium chrysogenum ? + succinate + CO2
-
 ?
1.14.11.26 deacetoxycephalosporin C + 2-oxoglutarate + O2
-
 Acremonium chrysogenum deacetylcephalosporin C + succinate + CO2
-
 ?
1.14.11.26 phenylacetyl-7-aminodeacetoxycephalosporanic acid + 2-oxoglutarate + O2
-
 Acremonium chrysogenum phenylacetyl-7-aminodeacetylcephalosporin + succinate + CO2
-
 ?
1.14.20.1 6-alpha-methylpenicillin N + 2-oxoglutarate + O2
-
 Acremonium chrysogenum ? + succinate + CO2 + H2O
-
 ?
1.14.20.1 6-alpha-methylpenicillin N + 2-oxoglutarate + O2 best substrate in the ring expansion reaction Acremonium chrysogenum ? + succinate + CO2 + H2O
-
 ir
1.14.20.1 6-aminopenicillanic acid + 2-oxoglutarate + O2
-
 Acremonium chrysogenum ? + succinate + CO2 + H2O
-
 ?
1.14.20.1 6-aminopenicillanic acid + 2-oxoglutarate + O2 low activity, ring expansion reaction Acremonium chrysogenum ? + succinate + CO2 + H2O
-
 ir
1.14.20.1 7-aminodeacetoxycephalosporanic acid + 2-oxoglutarate + O2 low activity, hydroxylation reaction Acremonium chrysogenum ? + succinate + CO2 + H2O
-
 ir
1.14.20.1 acetyl-6-aminopenicillanic acid + 2-oxoglutarate + O2
-
 Acremonium chrysogenum ? + succinate + CO2 + H2O
-
 ?
1.14.20.1 acetyl-6-aminopenicillanic acid + 2-oxoglutarate + O2 ring expansion reaction Acremonium chrysogenum ? + succinate + CO2 + H2O
-
 ir
1.14.20.1 adipyl-6-aminopenicillanic acid + 2-oxoglutarate + O2
-
 Acremonium chrysogenum ? + succinate + CO2 + H2O
-
 ?
1.14.20.1 adipyl-6-aminopenicillanic acid + 2-oxoglutarate + O2 ring expansion reaction Acremonium chrysogenum ? + succinate + CO2 + H2O
-
 ir
1.14.20.1 amoxicillin + 2-oxoglutarate + O2
-
 Acremonium chrysogenum ? + succinate + CO2 + H2O
-
 ?
1.14.20.1 amoxicillin + 2-oxoglutarate + O2 low activity, ring expansion reaction Acremonium chrysogenum ? + succinate + CO2 + H2O
-
 ir
1.14.20.1 ampicillin + 2-oxoglutarate + O2
-
 Acremonium chrysogenum ? + succinate + CO2 + H2O
-
 ?
1.14.20.1 ampicillin + 2-oxoglutarate + O2 ring expansion reaction Acremonium chrysogenum ? + succinate + CO2 + H2O
-
 ir
1.14.20.1 cephalexin + 2-oxoglutarate + O2 low activity, hydroxylation reaction Acremonium chrysogenum ? + succinate + CO2 + H2O
-
 ir
1.14.20.1 deacetoxycephalosporin C + 2-oxoglutarate + O2
-
 Acremonium chrysogenum deacetylcephalosporin C + succinate + CO2
-
 ir
1.14.20.1 deacetoxycephalosporin C + 2-oxoglutarate + O2 the penicillin /cephem substrate are bound by residues R161 and R163 Acremonium chrysogenum deacetylcephalosporin C + succinate + CO2
-
 ir
1.14.20.1 additional information deacetoxycephem substrate specificity for the hydroxylation reaction, penam substrate specificity of the bifunctional enzyme for the ring-expansion reation, overview Acremonium chrysogenum ?
-
 ?
1.14.20.1 penicillin G + 2-oxoglutarate + O2 the penicillin /cephem substrates are bound by residues R161 and R163 Acremonium chrysogenum phenylacetyl-7-aminodeacetoxycephalosporanic acid + succinate + CO2 + H2O
-
 ir
1.14.20.1 penicillin G + 2-oxoglutarate + O2
-
 Acremonium chrysogenum ? + succinate + CO2 + H2O
-
 ?
1.14.20.1 penicillin N + 2-oxoglutarate + O2
-
 Acremonium chrysogenum deacetoxycephalosporin C + succinate + CO2 + H2O
-
 ?
1.14.20.1 penicillin N + 2-oxoglutarate + O2
-
 Acremonium chrysogenum deacetoxycephalosporin C + succinate + CO2 + H2O
-
 ir
1.14.20.1 penicillin N + 2-oxoglutarate + O2 2-oxoglutarate is required, residues M181, R259, and S261 might be responsible for binding of 2-oxoglutarate, the penicillin /cephem substrate are bound by residues R161 and R163 Acremonium chrysogenum deacetoxycephalosporin C + succinate + CO2 + H2O
-
 ir
1.14.20.1 penicillin V + 2-oxoglutarate + O2
-
 Acremonium chrysogenum ? + succinate + CO2 + H2O
-
 ?
1.14.20.1 penicillin V + 2-oxoglutarate + O2 low activity, ring expansion reaction Acremonium chrysogenum ? + succinate + CO2 + H2O
-
 ir
1.14.20.1 phenyl-7-aminodeacetoxycephalosporanic acid + 2-oxoglutarate + O2 hydroxylation reaction Acremonium chrysogenum ? + succinate + CO2 + H2O
-
 ir

Subunits

EC Number Subunits Comment Organism
1.14.11.26 More structural model of enzyme Acremonium chrysogenum
1.14.20.1 ? x * 36348, wild-type enzyme, electrospray mass spectrometry Acremonium chrysogenum
1.14.20.1 More structural model of enzyme Acremonium chrysogenum
1.14.20.1 More MW of mutant enzyme determined by electrospray mass spectrometry Acremonium chrysogenum

Synonyms

EC Number Synonyms Comment Organism
1.14.20.1 DAOC/DACS
-
 Acremonium chrysogenum
1.14.20.1 deacetoxycephalosporin/deacetylcephalosporin C synthase
-
 Acremonium chrysogenum

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.14.20.1 30
-
 assay at Acremonium chrysogenum

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.14.20.1 0.016
-
 penicillin G 30°C, mutant M306I Acremonium chrysogenum
1.14.20.1 0.07
-
 penicillin G 30°C, triple mutant with truncation at residue 310, M306I and N305L Acremonium chrysogenum
1.14.20.1 0.073
-
 penicillin G pH 7.5, 30°C, recombinant wild-type enzyme Acremonium chrysogenum
1.14.20.1 0.073
-
 penicillin G 30°C, wild-type Acremonium chrysogenum
1.14.20.1 0.079
-
 penicillin G 30°C, mutant N305L Acremonium chrysogenum
1.14.20.1 0.091
-
 penicillin G 30°C, double mutant with truncation at residue 310 and M306I Acremonium chrysogenum

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.14.20.1 7.5
-
 assay at Acremonium chrysogenum