Information on EC 1.14.20.1 - deacetoxycephalosporin-C synthase

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The expected taxonomic range for this enzyme is: Eukaryota, Bacteria

EC NUMBER
COMMENTARY hide
1.14.20.1
-
RECOMMENDED NAME
GeneOntology No.
deacetoxycephalosporin-C synthase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
penicillin N + 2-oxoglutarate + O2 = deacetoxycephalosporin C + succinate + CO2 + H2O
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydroxylation
oxidation
-
-
-
-
redox reaction
-
-
-
-
reduction
-
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
deacetylcephalosporin C biosynthesis
-
-
Penicillin and cephalosporin biosynthesis
-
-
Metabolic pathways
-
-
Biosynthesis of antibiotics
-
-
SYSTEMATIC NAME
IUBMB Comments
penicillin-N,2-oxoglutarate:oxygen oxidoreductase (ring-expanding)
Forms part of the penicillin biosynthesis pathway (for pathway, click here).
CAS REGISTRY NUMBER
COMMENTARY hide
85746-10-7
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
formerly Cephalosporium acremonium, gene cefEF
SwissProt
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
metabolism
physiological function
additional information
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
(2S,5R,6R)-3,3-dimethyl-7-oxo-6-[(thiophen-2-ylacetyl)amino]-4-thia-1-azabicyclo[3.2.0]heptane-2-carboxylic acid + 2-oxoglutarate + O2
(6R,7R)-3-methyl-8-oxo-7-[(thiophen-2-ylacetyl)amino]-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid + succinate + CO2 + H2O
show the reaction diagram
-
-
-
-
?
3-exomethylenecephalosporin C + 2-oxoglutarate + O2
deacetylcephalosporin C + succinate + CO2 + H2O
show the reaction diagram
-
8% of the activity with cephalosporin N, recombinant enzyme
-
?
6-alpha-methylpenicillin N + 2-oxoglutarate + O2
? + succinate + CO2 + H2O
show the reaction diagram
6-aminopenicillanic acid + 2-oxoglutarate + O2
? + succinate + CO2 + H2O
show the reaction diagram
7-aminodeacetoxycephalosporanic acid + 2-oxoglutarate + O2
? + succinate + CO2 + H2O
show the reaction diagram
low activity, hydroxylation reaction
-
-
ir
acetyl-6-aminopenicillanic acid + 2-oxoglutarate + O2
7-acetylaminodesacetoxycephalosporanic acid + succinate + CO2 + H2O
show the reaction diagram
-
-
-
-
?
acetyl-6-aminopenicillanic acid + 2-oxoglutarate + O2
? + succinate + CO2 + H2O
show the reaction diagram
adipyl-6-aminopenicillanic acid + 2-oxoglutarate + O2
7-adipylaminodesacetoxycephalosporanic acid + succinate + CO2 + H2O
show the reaction diagram
-
-
-
-
?
adipyl-6-aminopenicillanic acid + 2-oxoglutarate + O2
7-aminodeacetoxycephalosporanic acid + succinate + CO2 + H2O
show the reaction diagram
-
poor substrate
-
?
adipyl-6-aminopenicillanic acid + 2-oxoglutarate + O2
? + succinate + CO2 + H2O
show the reaction diagram
amoxicillin + 2-oxoglutarate + O2
?
show the reaction diagram
amoxicillin + 2-oxoglutarate + O2
? + succinate + CO2 + H2O
show the reaction diagram
ampicillin + 2-oxo-4-methyl-pentanoic acid + O2
?
show the reaction diagram
-
mutants R258H, R258K, R258Q, R258A, R258L, R258F, no activity with the wild-type enzyme
-
-
ir
ampicillin + 2-oxo-4-methylpentanoate + O2
cephalexin + succinate + CO2 + H2O
show the reaction diagram
-
wild-type enzyme has a requirement for 2-oxoglutarate and cannot efficiently use hydrophobic 2-oxoacids, the mutants R258A, R258L, R258H and R258F have broadened cosubstrate selectivity and are able to utilize hydrophobic 2-oxoacids
-
?
ampicillin + 2-oxoglutarate + O2
?
show the reaction diagram
ampicillin + 2-oxoglutarate + O2
? + succinate + CO2
show the reaction diagram
-
-
-
-
?
ampicillin + 2-oxoglutarate + O2
? + succinate + CO2 + H2O
show the reaction diagram
ampicillin + 2-oxoglutarate + O2
cephalexin + succinate + CO2 + H2O
show the reaction diagram
ampicillin + 2-oxohexanoate + O2
cephalexin + ?
show the reaction diagram
-
wild-type enzyme has a requirement for 2-oxoglutarate and cannot efficiently use hydrophobic 2-oxoacids, the mutants R258A, R258L, R258H and R258F have broadened cosubstrate selectivity and are able to utilize hydrophobic 2-oxoacids
-
-
?
ampicillin + 2-oxohexanoic acid + O2
?
show the reaction diagram
-
mutants R258H, R258K, R258Q, R258A, R258L, R258F, no activity with the wild-type enzyme
-
-
ir
butyryl-6-aminopenicillanic acid + 2-oxoglutarate + O2
7-butyrylaminocephalosporanic acid + succinate + CO2 + H2O
show the reaction diagram
-
-
-
-
?
carbenicillin + 2-oxoglutarate + O2
?
show the reaction diagram
carbenicillin + 2-oxoglutarate + O2
? + succinate + CO2
show the reaction diagram
-
-
-
-
?
carbenicillin + 2-oxoglutarate + O2
? + succinate + CO2 + H2O
show the reaction diagram
cephalexin + 2-oxoglutarate + O2
? + succinate + CO2 + H2O
show the reaction diagram
low activity, hydroxylation reaction
-
-
ir
D-carboxymethylcysteinyl-6-aminopenicillanic acid + 2-oxoglutarate + O2
?
show the reaction diagram
-
-
-
-
?
deacetoxycephalosporin C + 2-oxoglutarate + O2
deacetylcephalosporin C + succinate + CO2
show the reaction diagram
deacetoxycephalosporin C + 2-oxoglutarate + O2
deacetylcephalosporin C + succinate + CO2 + H2O
show the reaction diagram
-
-
-
-
?
decanoyl-6-aminopenicillanic acid + 2-oxoglutarate + O2
7-decanoylaminodesacetoxycephalosporanic acid + succinate + CO2 + H2O
show the reaction diagram
-
-
-
-
?
heptanoyl-6-aminopenicillanic acid + 2-oxoglutarate + O2
7-heptanoylaminodesacetoxycephalosporanic acid + succinate + CO2 + H2O
show the reaction diagram
-
-
-
-
?
hexanoyl-6-aminopenicillanic acid + 2-oxoglutarate + O2
7-hexanoylaminodesacetoxycephalosporanic acid + succinate + CO2 + H2O
show the reaction diagram
-
-
-
-
?
metampicillin + 2-oxoglutarate + O2
?
show the reaction diagram
metampicillin + 2-oxoglutarate + O2
? + succinate + CO2 + H2O
show the reaction diagram
N-((thiophen-2-yl)acetyl)-6-aminopenicillanic acid + 2-oxoglutarate + O2
N-[(5R,6R)-3,3-dimethyl-2,7-dioxo-4-thia-1-azabicyclo[3.2.0]hept-6-yl]-2-(thiophen-2-yl)acetamide + succinate + CO2 + H2O
show the reaction diagram
-
-
-
-
?
N-acetyl-6-aminopenicillanic acid + 2-oxoglutarate + O2
N-acetyldeacetoxycephalosporin C + succinate + CO2 + H2O
show the reaction diagram
-
-
-
-
?
N-adipyl-6-aminopenicillanic acid + 2-oxoglutarate + O2
N-adipyldeacetoxycephalosporin C + succinate + CO2 + H2O
show the reaction diagram
-
-
-
-
?
N-adipyl-6-aminopenicillanic acid + 2-oxoglutarate + O2
N7-adipylaminodeacetoxycephalosporanic acid + succinate + CO2 + H2O
show the reaction diagram
N-butyryl-6-aminopenicillanic acid + 2-oxoglutarate + O2
N-butyryldeacetoxycephalosporin C + succinate + CO2 + H2O
show the reaction diagram
-
-
-
-
?
N-decanoyl-6-aminopenicillanic acid + 2-oxoglutarate + O2
N-decanoyldeacetoxycephalosporin C + succinate + CO2 + H2O
show the reaction diagram
-
-
-
-
?
N-heptanoyl-6-aminopenicillanic acid + 2-oxoglutarate + O2
N-heptanoyldeacetoxycephalosporin C + succinate + CO2 + H2O
show the reaction diagram
-
-
-
-
?
N-hexanoyl-6-aminopenicillanic acid + 2-oxoglutarate + O2
N-hexanoyldeacetoxycephalosporin C + succinate + CO2 + H2O
show the reaction diagram
-
-
-
-
?
N-nonanoyl-6-aminopenicillanic acid + 2-oxoglutarate + O2
N-nonanoyldeacetoxycephalosporin C + succinate + CO2 + H2O
show the reaction diagram
-
-
-
-
?
N-nonanoyl-6-aminopenicillanic acid + 2-oxoglutarate + O2
N7-nonanoyldeacetoxycephalosporin C + succinate + CO2 + H2O
show the reaction diagram
-
-
-
-
?
N-octanoyl-6-aminopenicillanic acid + 2-oxoglutarate + O2
N-octanoyldeacetoxycephalosporin C + succinate + CO2 + H2O
show the reaction diagram
-
-
-
-
?
N-octanoyl-6-aminopenicillanic acid + 2-oxoglutarate + O2
N7-octanoyldeacetoxycephalosporin C + succinate + CO2 + H2O
show the reaction diagram
-
-
-
-
?
N-valeryl-6-aminopenicillanic acid + 2-oxoglutarate + O2
N-valeryldeacetoxycephalosporin C + succinate + CO2 + H2O
show the reaction diagram
-
-
-
-
?
N-valeryl-6-aminopenicillanic acid + 2-oxoglutarate + O2
N7-pentanoyldeacetoxycephalosporin C + succinate + CO2 + H2O
show the reaction diagram
-
-
-
-
?
penicillin F + 2-oxoglutarate + O2
7-((3E)-hex-3-enoyl)aminocephalosporanic acid + succinate + CO2 + H2O
show the reaction diagram
-
-
-
-
?
penicillin F + 2-oxoglutarate + O2
?
show the reaction diagram
-
-
-
-
?
penicillin G + 2-oxo-4-methyl-pentanoic acid + O2
phenylacetyl-7-aminodeacetoxycephalosporanic acid + 3-methylbutanoate + CO2 + H2O
show the reaction diagram
-
mutants R258H, R258K, R258Q, R258A, R258L, R258F, no activity with the wild-type enzyme
-
-
ir
penicillin G + 2-oxo-4-methylpentanoate + O2
phenylacetyl-7-aminodeacetoxy cephalosporanic acid + ?
show the reaction diagram
penicillin G + 2-oxoglutarate + O2
7-aminodeacetoxycephalosporanate + succinate + CO2 + H2O
show the reaction diagram
penicillin G + 2-oxoglutarate + O2
7-phenylacetylaminocephalosporanic acid + succinate + CO2 + H2O
show the reaction diagram
-
-
-
-
?
penicillin G + 2-oxoglutarate + O2
?
show the reaction diagram
-
low catalytic effciency towards penicillin G
-
-
?
penicillin G + 2-oxoglutarate + O2
? + succinate + CO2
show the reaction diagram
-
-
-
-
?
penicillin G + 2-oxoglutarate + O2
? + succinate + CO2 + H2O
show the reaction diagram
-
-
-
?
penicillin G + 2-oxoglutarate + O2
phenylacetyl-7-aminodeacetoxy cephalosporanic acid + succinate + CO2 + H2O
show the reaction diagram
penicillin G + 2-oxoglutarate + O2
phenylacetyl-7-aminodeacetoxycephalosporanic acid + succinate + CO2 + H2O
show the reaction diagram
penicillin G + 2-oxohexanoate + O2
phenylacetyl-7-aminodeacetoxy cephalosporanic acid + ?
show the reaction diagram
penicillin G + 2-oxohexanoic acid + O2
phenylacetyl-7-aminodeacetoxycephalosporanic acid + pentanoate + CO2 + H2O
show the reaction diagram
-
mutants R258H, R258K, R258Q, R258A, R258L, R258F, no activity with the wild-type enzyme
-
-
ir
penicillin mX + 2-oxoglutarate + O2
7-[(3-hydroxyphenyl)acetyl]aminocephalosporanic acid + succinate + CO2 + H2O
show the reaction diagram
-
-
-
-
?
penicillin mX + 2-oxoglutarate + O2
?
show the reaction diagram
-
-
-
-
?
penicillin N + 2-oxoglutarate + O2
deacetoxycephalosporin C + succinate + CO2 + H2O
show the reaction diagram
penicillin V + 2-oxoglutarate + O2
?
show the reaction diagram
penicillin V + 2-oxoglutarate + O2
? + succinate + CO2 + H2O
show the reaction diagram
penicillin X + 2-oxoglutarate + O2
7-[(4-hydroxyphenyl)acetyl]aminocephalosporanic acid + succinate + CO2 + H2O
show the reaction diagram
-
-
-
-
?
penicillin X + 2-oxoglutarate + O2
?
show the reaction diagram
-
-
-
-
?
phenethicillin + 2-oxoglutarate + O2
?
show the reaction diagram
phenethicillin + 2-oxoglutarate + O2
? + succinate + CO2
show the reaction diagram
-
-
-
-
?
phenethicillin + 2-oxoglutarate + O2
? + succinate + CO2 + H2O
show the reaction diagram
phenyl-7-aminodeacetoxycephalosporanic acid + 2-oxoglutarate + O2
? + succinate + CO2 + H2O
show the reaction diagram
hydroxylation reaction
-
-
ir
ticarcillin + 2-oxoglutarate + O2
?
show the reaction diagram
-
-
-
-
?
additional information
?
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
(2S,5R,6R)-3,3-dimethyl-7-oxo-6-[(thiophen-2-ylacetyl)amino]-4-thia-1-azabicyclo[3.2.0]heptane-2-carboxylic acid + 2-oxoglutarate + O2
(6R,7R)-3-methyl-8-oxo-7-[(thiophen-2-ylacetyl)amino]-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid + succinate + CO2 + H2O
show the reaction diagram
-
-
-
-
?
acetyl-6-aminopenicillanic acid + 2-oxoglutarate + O2
7-acetylaminodesacetoxycephalosporanic acid + succinate + CO2 + H2O
show the reaction diagram
-
-
-
-
?
adipyl-6-aminopenicillanic acid + 2-oxoglutarate + O2
7-adipylaminodesacetoxycephalosporanic acid + succinate + CO2 + H2O
show the reaction diagram
-
-
-
-
?
amoxicillin + 2-oxoglutarate + O2
?
show the reaction diagram
ampicillin + 2-oxoglutarate + O2
?
show the reaction diagram
butyryl-6-aminopenicillanic acid + 2-oxoglutarate + O2
7-butyrylaminocephalosporanic acid + succinate + CO2 + H2O
show the reaction diagram
-
-
-
-
?
carbenicillin + 2-oxoglutarate + O2
?
show the reaction diagram
D-carboxymethylcysteinyl-6-aminopenicillanic acid + 2-oxoglutarate + O2
?
show the reaction diagram
-
-
-
-
?
deacetoxycephalosporin C + 2-oxoglutarate + O2
deacetylcephalosporin C + succinate + CO2
show the reaction diagram
deacetoxycephalosporin C + 2-oxoglutarate + O2
deacetylcephalosporin C + succinate + CO2 + H2O
show the reaction diagram
-
-
-
-
?
decanoyl-6-aminopenicillanic acid + 2-oxoglutarate + O2
7-decanoylaminodesacetoxycephalosporanic acid + succinate + CO2 + H2O
show the reaction diagram
-
-
-
-
?
heptanoyl-6-aminopenicillanic acid + 2-oxoglutarate + O2
7-heptanoylaminodesacetoxycephalosporanic acid + succinate + CO2 + H2O
show the reaction diagram
-
-
-
-
?
hexanoyl-6-aminopenicillanic acid + 2-oxoglutarate + O2
7-hexanoylaminodesacetoxycephalosporanic acid + succinate + CO2 + H2O
show the reaction diagram
-
-
-
-
?
metampicillin + 2-oxoglutarate + O2
?
show the reaction diagram
-
-
-
-
?
N-adipyl-6-aminopenicillanic acid + 2-oxoglutarate + O2
N7-adipylaminodeacetoxycephalosporanic acid + succinate + CO2 + H2O
show the reaction diagram
N-nonanoyl-6-aminopenicillanic acid + 2-oxoglutarate + O2
N7-nonanoyldeacetoxycephalosporin C + succinate + CO2 + H2O
show the reaction diagram
-
-
-
-
?
N-octanoyl-6-aminopenicillanic acid + 2-oxoglutarate + O2
N7-octanoyldeacetoxycephalosporin C + succinate + CO2 + H2O
show the reaction diagram
-
-
-
-
?
N-valeryl-6-aminopenicillanic acid + 2-oxoglutarate + O2
N7-pentanoyldeacetoxycephalosporin C + succinate + CO2 + H2O
show the reaction diagram
-
-
-
-
?
penicillin F + 2-oxoglutarate + O2
7-((3E)-hex-3-enoyl)aminocephalosporanic acid + succinate + CO2 + H2O
show the reaction diagram
-
-
-
-
?
penicillin G + 2-oxoglutarate + O2
7-phenylacetylaminocephalosporanic acid + succinate + CO2 + H2O
show the reaction diagram
-
-
-
-
?
penicillin G + 2-oxoglutarate + O2
phenylacetyl-7-aminodeacetoxycephalosporanic acid + succinate + CO2 + H2O
show the reaction diagram
penicillin mX + 2-oxoglutarate + O2
7-[(3-hydroxyphenyl)acetyl]aminocephalosporanic acid + succinate + CO2 + H2O
show the reaction diagram
-
-
-
-
?
penicillin N + 2-oxoglutarate + O2
deacetoxycephalosporin C + succinate + CO2 + H2O
show the reaction diagram
penicillin V + 2-oxoglutarate + O2
?
show the reaction diagram
-
-
-
-
?
penicillin X + 2-oxoglutarate + O2
7-[(4-hydroxyphenyl)acetyl]aminocephalosporanic acid + succinate + CO2 + H2O
show the reaction diagram
-
-
-
-
?
phenethicillin + 2-oxoglutarate + O2
?
show the reaction diagram
-
-
-
-
?
ticarcillin + 2-oxoglutarate + O2
?
show the reaction diagram
-
-
-
-
?
additional information
?
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
(2S,4S,6R,7R)-1-aza-7-((5R)-5-carboxypentanamidol)-4-methyl-8-oxo-5-thiatricyclo-(4,2,0,0)octane-2-carboxylate
-
reversible, 0.04 mM, 90% inhibition
1,10-phenanthroline
-
-
2-oxobutyrate
-
0.1 mM, complete inhibition
3-exomethylenecephalosporin
-
strong, substrate for hydroxylation reaction of enzyme, EC 1.14.11.26
-
3-exomethylenecephalosporin C
-
0.28 mM, 71% inhibition of the formation of deacetoxycephalosporin C
3-oxoadipate
-
0.1 mM, 56% inhibition
5,5'-dithiobis-2-nitrobenzoic acid
ammonium hydrogencarbonate
-
100-500 mM, complete inactivation
ampicillin
-
; 2.8 mM, 13% inhibition of the formation of deacetoxycephalosporin C
bathophenanthroline
-
-
Cu2+
-
1 mM, complete inhibition
iodoacetic acid
N-ethylmaleimide
Ni2+
-
inhibition in decreasing order, Zn2+, Co2+, Ni2+
o-phenanthroline
p-hydroxymercuribenzoate
penicillin G
-
; 2.8 mM, 57% inhibition of the formation of deacetoxycephalosporin C
Penicillin V
-
; 2.8 mM, 47% inhibition of the formation of deacetoxycephalosporin C
additional information
-
when 2-oxoglutarate is added to the enzyme before the mixture of ATP, Fe2+ and ascobate activity is only about 40% of the reaction
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
ammonium chloride
-
-
ammonium sulfate
-
-
ascorbate
bovine serum albumin
-
stimulates
-
catalase
-
stimulates
-
dithiothreitol
DTT
-
reducing agent stimulates DAOC synthase activity
Fe2+
-
requires the Fe2+ as a cofactor
glutathione
-
reduced
additional information
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.003 - 0.022
2-oxoglutarate
3.28
acetyl-6-aminopenicillanic acid
-
wild type enzyme
1.3
adipyl 6-aminopenicillanic acid
-
-
0.05 - 24
ampicillin
0.028
deacetoxycephalosporin C
-
-
0.003
oxoglutarate
-
-
0.014 - 48.1
penicillin G
0.004 - 0.295
Penicillin N
additional information
additional information
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.06
acetyl-6-aminopenicillanic acid
-
-
0.073
adipyl 6-aminopenicillanic acid
-
-
0.005 - 0.122
ampicillin
0.001 - 0.668
penicillin G
0.02 - 0.42
Penicillin N
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.003 - 0.736
penicillin G
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.034
3-exomethylenecephalosporin C
-
-
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.013
recombinant enzyme, substrate amoxicillin
0.014
purified recombinant wild-type and truncated mutant enzyme, substrate metampicillin
0.015
-
recombinant enzyme, substrate metampicillin
0.023
purified recombinant truncated mutant enzyme, substrate carbenicillin
0.037
-
recombinant enzyme, substrate carbenicillin
0.039
purified recombinant wild-type and truncated mutant enzyme, substrate ampicillin, determined by bioassay
0.051
recombinant enzyme, substrate penicillin V
0.071
purified recombinant truncated mutant enzyme, substrate phenethicillin
0.074
purified recombinant wild-type enzyme, substrate phenethicillin
0.128
recombinant enzyme, substrate carbenicillin
0.133
purified recombinant wild-type enzyme, substrate ampicillin, determined by HPLC
0.156
purified recombinant wild-type enzyme, substrate 6-alpha-methylpenicillin N, substrate conversion
0.166
-
native enzyme
0.178
purified recombinant truncated mutant enzyme, substrate ampicillin, determined by HPLC
0.203
-
recombinant enzyme, substrate ampicillin, determined by bioassay
0.237
purified recombinant truncated mutant enzyme, substrate penicillin G
0.24
purified recombinant wild-type enzyme, substrate penicillin G
0.36
-
recombinant enzyme, substrate penicillin G
0.411
-
recombinant enzyme, substrate phenethicillin
0.432
-
recombinant enzyme
0.458
recombinant enzyme, substrate ampicillin, determined by bioassay
0.76
-
substrate carbenicillin, wild-type, pH 7.4, 30°C
0.827
-
pH 7.5, 30°C
0.932
recombinant enzyme, substrate ampicillin; recombinant enzyme, substrate ampicillin, determined by HPLC
1.042
recombinant enzyme, substrate penicillin G
1.551
recombinant enzyme, substrate phenethicillin
2.221
-
recombinant enzyme, substrate ampicillin, determined by HPLC
3.57
-
substrate phenethicillin, wild-type, pH 7.4, 30°C
5.67
-
substrate ampicillin, wild-type, pH 7.4, 30°C
6.41
-
substrate ampicillin, wild-type, 30°C
6.72
-
substrate penicillin G, wild-type, pH 7.4, 30°C
7.23
-
substrate ampicillin, mutant R306M, 30°C
9.11
-
substrate ampicillin, mutant R306I, 30°
10.9
-
substrate ampicillin, mutant R306L, 30°C
11.89
-
recombinant mutant S261I, pH not specified in the publication, 30°C
13.44
-
recombinant mutant C37S, pH not specified in the publication, 30°C
16.87
-
recombinant mutant T42A, pH not specified in the publication, 30°C
20.03
-
recombinant mutant A61E, pH not specified in the publication, 30°C
22.06
-
recombinant mutant S261V, pH not specified in the publication, 30°C
30.86
-
recombinant mutant S261L, pH not specified in the publication, 30°C
31.38
-
recombinant mutant Y184L, pH not specified in the publication, 30°C
33.51
-
recombinant mutant Y184M, pH not specified in the publication, 30°C
36.12
-
recombinant mutant Y184I, pH not specified in the publication, 30°C
38.5
-
pH 7.4, 27°C
49.12
-
recombinant mutant S59T, pH not specified in the publication, 30°C
56.84
-
recombinant wild-type enzyme, pH not specified in the publication, 30°C
66
-
recombinant enzyme
71.7
-
recombinant mutant Q126M, pH not specified in the publication, 30°C
80.68
-
recombinant mutant T213V, pH not specified in the publication, 30°C
90.04
-
recombinant mutant S261M, pH not specified in the publication, 30°C
98
-
recombinant mutant S261A, pH not specified in the publication, 30°C
160
-
recombinant mutant Y184A, pH not specified in the publication, 30°C
190
-
purified recombinant wild-type
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.5
-
assay at, substrate penicillin G
7
-
; in piperazineethanesulfonic acid buffer, recombinant enzyme
7.4
-
native enzyme
7.5 - 7.8
-
expandase reaction, EC 1.14.20.1
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
26 - 34
-
expandase reaction, EC 1.14.20.1
36
-
; native and recombinant enzyme
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5 - 40
-
20°C: 15% of maximal activity, 35°C: 15% of maximal activity
20 - 35
-
20°C: 90% of maximal activity, 35°C: 90% of maximal activity
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.3
-
isoelectric focusing
6.1
-
isoelectric focusing
6.3
-
isoelectric focusing
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
PDB
SCOP
CATH
UNIPROT
ORGANISM
Streptomyces clavuligerus (strain ATCC 27064 / DSM 738 / JCM 4710 / NBRC 13307 / NCIMB 12785 / NRRL 3585 / VKM Ac-602);
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
27000
-
gel filtration
28000
-
1 * 28000, SDS-PAGE
29500
-
gel filtration
30966
-
3 * 30966, DAOCS is in equilibrium with a trimeric form which is the form that crystallizes
34500
-
x * 34500, about, recombinant wild-type and mutant enzymes, SDS-PAGE
34519
-
x * 34519, calculation from nucleotide sequence; x * 36000, SDS-PAGE, x * 34519, calculated
34554
-
x * 34554, in solution the monomeric apoenzyme is in equilibrium with atrimeric form, electrospray mass spectrometry
35000
-
gel filtration
36000
-
x * 36000, SDS-PAGE; x * 36000, SDS-PAGE, x * 34519, calculated
36348
x * 36348, wild-type enzyme, electrospray mass spectrometry
36642
-
x * 40000, SDS-PAGE, x * 36642, calculated
41000
-
1 * 41000, SDS-PAGE
43000
-
gel filtration
62000
x * 60000, recombinant truncated mutant enzyme, SDS-PAGE, x * 62000, recombinant wild-type enzyme
additional information
-
gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
monomer
trimer
additional information
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
crystal structure of the DAOC synthase is in trimeric form, C-terminal end of the protein is responsible for the oligomerization process, addition of Fe2+ or 2-oxoglutarate shifts the equilibrium toward the monomeric form that appears to be the active form in solution
-
; as apoprotein, 1.3 A resolution; as apoprotein with N-terminal His tag, 2.3 A resolution; crystallization of mutant delta R306, with Fe2+ and 2-oxoglutarate, 2.1 A resolution; crystallization of mutant delta R307A, with Fe2+, succinate and CO2, 1.96 A resolution; crystallization with 5-hydroxy-4-ketovaleric acid, 1.53 A resolution; crystallization with Fe2+, 1.5 A resolution; crystallization with Fe2+ and 2-oxoglutarate, 1.5 A resolution, the crystal structure of scDAOCS complexed with 2-oxoglutarate reveals that the 5-carboxylate of 2-oxoglutarate is stabilized by electrostatic interaction with the side chain of R258; crystallization with Fe(II), 2-oxoglutarate and ampicillin, 1.5 A resolution; crystallization with Fe(II), 2-oxoglutarate and penicillin G, 1.7 A resolution; crystallization with Fe(II) and deacetoxycephalosporin C, 1.7 A resolution; crystallization with Fe(II) and penicillin G, 1.6 A resolution; crystallization with Fe(II) and succinate , 1.5 A resolution; R258Q mutant, crystallization with Fe(II) and alpha-keto-beta-methylbutanoate, 1.5 and 1.6 A resolution; with N-terminal His tag and Fe2+, 2.51 A resolution; with N-terminal His tag, crystallization with ampicillin and Fe2+, 2.7 A resolution; with N-terminal His tag, crystallization with deacetoxycephalosporin C and Fe2+, 3.0 A resolution
-
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
-
hanging drop method, recombinant enzyme expressed in Escherichia coli
-
quantum mechanical calculations of the first part of the reaction based on the high-resolution structures of the active site and its complexes with ligands
-
recombinant enzyme expressed in Escherichia coli, high-resolution structures for apoenzyme, the enzyme complexed with Fe(II), and with Fe(II) and 2-oxoglutarate
-
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
-
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.5 - 9
-
; 4°C, 1 h, stable, native enzyme
440380
7
-
4°C, 120 h, 80% loss of activity
440378
8
-
4°C, 120 h, 70% loss of activity
440378
9
-
4°C, 120 h, 35% loss of activity
440378
10
-
enzyme has similar stability at 4°C and at 25°C
440375
11
-
enzyme is more stable at 4°C than at 25°C
440375
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
25
-
60 min, stable
30
-
60 min, 25% loss of activity
40
-
10 min, 50% loss of activity; stable up to
65
-
60 min, complete inactivation
additional information
-
at pH 10 enzyme has similar stability at 4°C and at 25°C. At pH 11 the enzyme is more stable at 4°C than at 25°C
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
increasing ionic strength to 100 decreases stability of the enzyme
-
purified enzyme is stable when flash-frozen and stored at -80°C
-
sodium phosphate and Bis/Tris buffers are inhibitory
-
the enzyme is very unstable and can be partially stabilized in 25 mM Tris-HCl, pH 9.0, in the presence of 0.1 mM DTT
-
ORGANIC SOLVENT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
urea
-
purification is achieved by an anion-exchange-chromatographic step in the presence of denaturing concentrations of urea. The main obstacle to converting the homogenous unfolded protein into the active enzyme is a urea-dependent aggregation during refolding that led to irreversible enzyme inactivation
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-70°C, presence of dithiothreitol, stable for 4 weeks
-
4°C, presence of dithiothreitol, half-life is 48 h
-
purified enzyme is stable when flash-frozen and stored at -80°C
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
; affinity chromatography
-
anion-exchange and gel filtration
-
efficient large-scale purification; recombinant enzyme expressed in Escherichia coli
-
expression in Escherichia coli, overexpression as an insoluble and inactive enzyme, elaboration of refolding scheme resulting in highliy active and moderately stable enzyme
-
native enzyme, partial, recombinant enzyme, to near homogenity; recombinant enzyme expressed in Escherichia coli
-
purification of partially folded recombinant expandase/hydroxylase from insoluble granules of recombinant Escherichia coli
-
purification of refolding-competent recombinant enzyme
-
recombinant enzyme expressed in Escherichia coli
-
recombinant enzyme from Escherichia coli
recombinant enzyme from Escherichia coli strain BL21(DE3) by affinity chromatography
recombinant N-terminally His-tagged wild-type and C-terminally truncated mutant enzymes from Escherichia coli strain BL21(DE3)
-
recombinant wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by anion exchange chromatography and gel filtration; recombinant wild-type enzyme and mutant enzymes R258K, R258H, R258A, R258L and R258F
-
recombinant wild-type and mutant enzymes from Escherichia coli strain ESS by ion exchange chromatography and gel filtration
-
soluble recombinant wild-type and mutant enzymes from Escherichia coli strain BL21 by anion exchange chromatography and gel filtration
-
wild-type and mutant enzymes R74I, R74Q, R75I/D270G, R75Q, R306L and R307Q
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
cloning and heterologous expression in Escherichia coli, Streptomyces lividans, Penicillium chrysogenum, Pseudomonas putida and Pichia pastoris, using of commonly used glutathione S-transferase to express DAOCS as a fusion protein; expressed in Escherichia coli, Streptomyces lividans, Penicillium chrysogenum, Pseudomonas putida, and Pichia pastoris; the attempt to improve the activity of DAOCS by the directed evolution approach is probably the construction of hybrid DAOCS of Streptomyces clavuligerus and Nocardia lactamdurans using in-vivo homologous recombination
-
DNA and amino acid sequence determination and analysis, expression as GST-fusion protein in Escherichia coli strain BL21(DE3)
DNA and amino acid sequence determination and analysis, expression of the wild-type enzyme and a C-terminally truncated mutant enzyme lacking 20 amino acids as GST-fusion proteins in Escherichia coli strain BL21(DE3)
expressed in Escherichia coli
expression as glutathione S-transferase fusion protein in Echerichia coli
-
expression at high levels in Escherichia coli under control of the trc promoter
-
expression in Escherichia coli
-
expression of wild-type and C-terminally truncated mutant enzymes as N-terminally His-tagged proteins in Escherichia coli strain BL21(DE3)
-
expression of wild-type and mutant enzymes at high levels in Escherichia coli
-
expression of wild-type and mutant enzymes in Escherichia coli strain BL21
-
expression of wild-type and mutant enzymes in Escherichia coli strain BL21(DE3)
-
expression of wild-type and mutant enzymes in Escherichia coli strain BL21(DE3), subcloning in strain XL-1 Blue
expression of wild-type and mutant enzymes in Escherichia coli strain ESS
-
gene cefE, recombinant expression in Escherichia coli BW25311. Recombinant expression of a hybrid enzyme,with parts from the