Information on EC 3.1.1.41 - cephalosporin-C deacetylase

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

EC NUMBER
COMMENTARY hide
3.1.1.41
-
RECOMMENDED NAME
GeneOntology No.
cephalosporin-C deacetylase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
cephalosporin C + H2O = deacetylcephalosporin C + acetate
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
carboxylic ester hydrolysis
hydrolysis of carboxylic ester
-
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
Biosynthesis of antibiotics
-
-
Penicillin and cephalosporin biosynthesis
-
-
SYSTEMATIC NAME
IUBMB Comments
cephalosporin-C acetylhydrolase
Hydrolyses the acetyl ester bond on the 10-position of the antibiotic cephalosporin C.
CAS REGISTRY NUMBER
COMMENTARY hide
52227-71-1
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
strain MA4213 and LC411
-
-
Manually annotated by BRENDA team
strain MA4213 and LC411
-
-
Manually annotated by BRENDA team
strain CECT 5072, gene axe
-
-
Manually annotated by BRENDA team
strain CECT 5072, gene axe
-
-
Manually annotated by BRENDA team
strain ATCC6633
-
-
Manually annotated by BRENDA team
gene Cah
UniProt
Manually annotated by BRENDA team
strain SHS 0133
-
-
Manually annotated by BRENDA team
strain SHS0133, recombinant enzyme from Escherichia coli immobilized on an anion exchange resin
-
-
Manually annotated by BRENDA team
strain WRRL-B-558
-
-
Manually annotated by BRENDA team
Citrus sp.
-
-
-
Manually annotated by BRENDA team
gene aes
-
-
Manually annotated by BRENDA team
strain AY F-298
-
-
Manually annotated by BRENDA team
strain AY F-298
-
-
Manually annotated by BRENDA team
GK-16
-
-
Manually annotated by BRENDA team
GK-16
-
-
Manually annotated by BRENDA team
strain 38B1
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
gene TM0077
UniProt
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
additional information
the active site has a classic catalytic triad, consisting of Ser188 as the nucleophile, His303 as the proton acceptor/donor, and Asp274 as the acidic residue stabilizing the histidine. The catalytic serine Ser188 is located within a conserved pentapeptide sequence, Gly-X-Ser-X-Gly (GGSQG), characteristic of esterases and lipases, overview
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
1,2,3,4-tetra-O--acetyl-xylopyranose + H2O
?
show the reaction diagram
-
-
?
2,3,2',3',4'-penta-acetyl-xylobiose + H2O
?
show the reaction diagram
-
-
?
2-naphthyl acetate + H2O
2-naphthol + acetate
show the reaction diagram
3'-O-acetyl-2'-deoxyadenosine + H2O
?
show the reaction diagram
-
-
?
3-naphthyl acetate + H2O
3-naphthol + acetate
show the reaction diagram
-
-
-
?
4-methylumbelliferyl acetate + H2O
4-methylumbelliferol + acetate
show the reaction diagram
-
-
-
?
4-nitrophenyl acetate + H2O
4-nitrophenol + acetate
show the reaction diagram
4-nitrophenyl beta-D-xylopyranoside + H2O
4-nitrophenol + D-xylopyranose
show the reaction diagram
4-nitrophenyl-beta-D-xylopyranoside monoacetates as substrates in a beta-xylosidase-coupled assay, TM0077 hydrolyzes acetate at positions 2, 3 and 4 with equal efficiencyas substrates in a ?-xylosidase-coupled assay
-
-
?
4-nitrophenyl butyrate + H2O
4-nitrophenol + butyrate
show the reaction diagram
4-nitrophenyl caprylate + H2O
4-nitrophenol + caprylate
show the reaction diagram
-
-
-
?
7-aminocephalosporanic acid + H2O
7-amino-3-deacetylcephalosporanic acid + acetate
show the reaction diagram
7-aminocephalosporanic acid + H2O
acetate + deacetyl-7-aminocephalosporanic acid
show the reaction diagram
7-aminocephalosporanic acid + H2O
deacetyl 7-aminocephalosporanic acid + acetate
show the reaction diagram
alpha-ketoadipinyl-7-aminocephalosporanic acid + H2O
7-aminocephalosporanic acid + alpha-ketoadipate
show the reaction diagram
-
-
-
-
-
alpha-naphthyl acetate + H2O
1-naphthol + acetate
show the reaction diagram
benzylcephalosporin + H2O
acetate + deacetylbenzylcephalosporin
show the reaction diagram
benzyloxycarbonyl-7-aminocephalosporanic acid + H2O
?
show the reaction diagram
cephaloglycin + H2O
acetate + (6R,7R)-7-{[(2R)-2-amino-2-phenylacetyl]amino}-3-(hydroxymethyl)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid
show the reaction diagram
cephalosporin C + H2O
deacetylcephalosporin C + acetate
show the reaction diagram
cephalothin + H2O
acetate + deacetylcephalothin
show the reaction diagram
cephapirin + H2O
acetate + deacetylcephapirin
show the reaction diagram
deacetyl-7-aminocephalosporanic acid + ethyl acetate
7-aminocephalosporanic acid + ethanol
show the reaction diagram
-
-
-
?
deacetyl-7-aminocephalosporanic acid + ethylene glycol diacetate
7-aminocephalosporanic acid + ?
show the reaction diagram
deacetyl-7-aminocephalosporanic acid + isopropenyl acetate
7-aminocephalosporanic acid + ?
show the reaction diagram
-
-
-
?
deacetyl-7-aminocephalosporanic acid + methyl acetate
7-aminocephalosporanic acid + methanol
show the reaction diagram
-
-
-
?
glucose pentaacetate + H2O
?
show the reaction diagram
glucose pentacetate + H2O
?
show the reaction diagram
-
-
-
?
glutaryl-7-aminocephalosporanic acid + H2O
7-aminocephalosporanic acid + glutamate
show the reaction diagram
monoacetin + H2O
acetate + glycerol
show the reaction diagram
-
-
-
-
?
monochloroacetyl-7-aminocephalosporanic acid + H2O
acetate + monochloroacetyl-deacetyl-7-aminocephalosporanic acid
show the reaction diagram
p-nitrophenyl acetate + H2O
4-nitrophenol + acetate
show the reaction diagram
phenoxymethylcephalosporin + H2O
acetate deacetylphenoxycephalosporin
show the reaction diagram
triacetin + H2O
?
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
cephalosporin C + H2O
deacetylcephalosporin C + acetate
show the reaction diagram
additional information
?
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Ca2+
not required for activity, one calcium ion is located at the N-terminal region of helix alphaA-1, and is coordinated by the backbone carbonyl of Lys22 and the Glu26 carboxylate, Asp25 carboxylate contributes to the calcium binding via one of the coordinating water molecules. Another calcium ion is bound in a crystal packing interface between chain A and chain C' of a crystallographic symmetry-related hexamer and is coordinated by the carboxylates of GluA45 and AspA58 from one chain and the carboxylate from Glu C'45 (bidentate coordination) of the symmetry-related chain with three water molecules completing a capped-octahedral coordination sphere
additional information
no significant stimulation or reduction of enzyme activity in the presence of divalent metal ions or EDTA. In the paraoxon-bound structure, the diethylphosphate moiety is stabilized by hydrogen-bonding interactions with the oxyanion hole. One of the two ethyl arms of bound paraoxon points toward the larger pocket in the protein, while the other follows the groove of the small pocket. The two ethyl arms are stabilized by packing against Tyr92, Trp124, Pro228, Ile276, and His303
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
(p-amino-phenyl)methanesulfonyl fluoride
-
slight inhibition
3,4-dichloroisocoumarin
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-
5,5'-dithiobis(2-nitrobenzoate)
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-
acetate
AsO43-
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40% inhibition at 1 mM
Ba2+
low inhibition at 1 mM
Cd2+
low inhibition at 1 mM
Co2+
low inhibition at 1 mM
deacetyl-7-aminocephalosporanic acid
diethyldicarbonate
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diisopropylfluorophosphate
dimethyl phosphite
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-
dimethylphosphite
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slight inhibition
EDTA
-
slight inhibition
eserine sulfate
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slight inhibition
Fe2+
low inhibition at 1 mM
Fe3+
strong inhibition at 1 mM
iodoacetamide
-
-
Mg2+
low inhibition at 1 mM
Mn2+
low inhibition at 1 mM
N-tosyl-L-Lys chloromethyl ketone
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-
Paraoxon
competitive irreversible inhibitors of esterases. Inhibition proceeds by the formation of a reversible Michaelis complex, followed by an irreversible step, the inhibitor binds covalently to the catalytic serine (Ser188), upon binding of inhibitor, the catalytic serine adopts an altered conformation
phenylmethylsulfonyl fluoride
SDS
low inhibition at 1 mM
trimethylphosphite
-
slight inhibition
additional information
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1.43 - 8.3
7-aminocephalosporanic acid
1.11
benzyloxycarbonyl-7-aminocephalosporanic acid
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-
9.2 - 11.5
cephaloglycin
4.7 - 215
cephalosporin C
8.3
Cephalothin
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-
8.6 - 11.7
cephalotin
13.9 - 16.7
cephapirin
2.03
monochloroacetyl-7-aminocephalosporanic acid
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-
1
p-nitrophenyl acetate
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-
0.46 - 32
Triacetin
additional information
additional information
-
kinetics
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
576
7-aminocephalosporanic acid
Rhodotorula glutinis
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-
534
benzyloxycarbonyl-7-aminocephalosporanic acid
Rhodotorula glutinis
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-
1370
cephalosporin C
Rhodotorula glutinis
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55
Cephalothin
Bacillus subtilis
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-
80
cephapirin
Bacillus subtilis
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-
768
monochloroacetyl-7-aminocephalosporanic acid
Rhodotorula glutinis
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-
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
additional information
additional information
inhibition kinetics
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SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
2.72
purified recombinant enzyme, pH 7.0, 40°C, substrate 4-nitrophenyl butyrate
187.4
-
hydrolysis of p-nitrophenyl acetate
741
purified recombinant enzyme, pH 7.0, 40°C, substrate alpha-naphthyl acetate
962
purified recombinant enzyme, pH 7.0, 40°C, substrate beta-naphthyl acetate
1086
purified recombinant enzyme, pH 7.0, 40°C, substrate 4-methylumbelliferyl acetate
1245
purified recombinant enzyme, pH 7.0, 40°C, substrate glucose pentacetate
2949
purified recombinant enzyme, pH 7.0, 40°C, substrate 4-nitrophenyl acetate
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6 - 6.5
Citrus sp.
-
hydrolysis of monoacetin
7.5 - 8
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hydrolysis of cephalothin
7.6
-
hydrolysis of cephalosporin C
8
-
strain ATCC6633
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4 - 11
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significant activity within this range
4.2 - 7
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pH 4.2: 30% of maximal activity, pH 7.0: about 35% of maximal activity
5 - 10
activity range, profile overview
5.5 - 7
Citrus sp.
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10% of maximal activity at pH 5.5 and at pH 7.0
6.5 - 8.3
-
pH 6.5: about 25% of maximal activity, pH 8.3: about 70% of maximal activity
6.5 - 9
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pH 6.5: about 65% of maximal activity, pH 9.0: about 95% of maximal activity
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5 - 65
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significant activity within this range
10 - 45
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10°C: about 40% of maximal activity, 45°C: about 30% of maximal activity
20 - 55
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20°C: about 30% of maximal activity, 55°C: about 10% of maximal activity
20 - 60
-
20°C: about 40% of maximal activity, 60°C: about 10% of maximal activity
30 - 75
activity range, profile overview
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4
-
isoelectric focusing, pH gradient 3-10
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
Citrus sp.
-
peel
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
the enzyme has no predicted signal sequence
Manually annotated by BRENDA team
PDB
SCOP
CATH
ORGANISM
UNIPROT
Bacillus subtilis (strain 168)
Bacillus subtilis (strain 168)
Bacillus subtilis (strain 168)
Thermoanaerobacterium saccharolyticum (strain DSM 8691 / JW/SL-YS485)
Thermotoga maritima (strain ATCC 43589 / MSB8 / DSM 3109 / JCM 10099)
Thermotoga maritima (strain ATCC 43589 / MSB8 / DSM 3109 / JCM 10099)
Thermotoga maritima (strain ATCC 43589 / MSB8 / DSM 3109 / JCM 10099)
Thermotoga maritima (strain ATCC 43589 / MSB8 / DSM 3109 / JCM 10099)
Thermotoga maritima (strain ATCC 43589 / MSB8 / DSM 3109 / JCM 10099)
Thermotoga maritima (strain ATCC 43589 / MSB8 / DSM 3109 / JCM 10099)
Thermotoga maritima (strain ATCC 43589 / MSB8 / DSM 3109 / JCM 10099)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
25000
-
sucrose density gradient centrifugation
31000
-
SDS-PAGE, gel filtration
35000
-
8 * 35000, strain SHS 0133, SDS-PAGE
36000
6 * 36000, recombinant enzyme, SDS-PAGE
38230
-
preprotein, calculated from amino acid sequence
44000
-
SDS-PAGE
61315
-
x * 61315, calculation from amino acid sequence
72100
-
gel filtration
80000
-
x * 80000, SDS-PAGE
82000
-
2 * 82000, SDS-PAGE
141000
-
gel filtration
150000
-
strain ATCC6633
190000
-
gel filtration
223000
recombinant enzyme, gel filtration
280000
-
strain SHS 0133, gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
?
-
x * 61315, calculation from amino acid sequence; x * 80000, SDS-PAGE
hexamer
octamer
tetramer
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
proteolytic modification
side-chain modification
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
purified recombinant selenomethionine-substituted and wild-type enzyme in complex with covalently bound phenylmethylsulfonyl fluoride and paraoxon, hanging drop vapour diffusion method, for selenomethionine-substituted enzyme: mixing of 500 nl of 15 mg/ml protein in 20 mM Tris pH 7.9, 150 mM NaCl, 0.25 mM TCEP, in presence or absence of inhibitors, with 500 nl of reservoir solution containing 20% w/v PEG-3000, 0.1 M HEPES pH 7.5, 0.2 M NaCl, and equilibration against 0.25 ml of reservoir solution, for wild-type enzyme: mixing of 100 nl of 22.8 mg/ml protein in 20 mM Tris pH 7.9, 150 mM NaCl, 0.25 mM TCEP, in presence or absence of inhibitors, with 100 nl of reservoir solution containing 0.2 M calcium acetate hydrate, 20% w/v PEG 3350, pH 7.3, and equilibration against 0.06 ml of reservoir solution, 20°C, X-ray diffraction structure determination and analysis at 2.4 A and 2.1 A resolution, respectively, molecular replacement, structure modeling
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4 - 11
-
immobilized enzyme stable for at least 60 min at 20°C
671502
4
Citrus sp.
-
rapid inactivation at or below
170668
4.5 - 12
purified recombinant enzyme, over 80% of maximal activity within this range
729073
5
-
relatively labile below
170671
5 - 8.3
Citrus sp.
-
25°C, stable
170668
7 - 10
-
37°C, 30 min, stable
170673
10
-
free enzyme shows highest activity after incubation at pH10 for 60 min at 20°C, about 15% of activity remaining after incubation at pH4
671502
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
40
purified recombinant enzyme, pH 7.0, stable up to, loss of 10% activity after 120 h
45
-
50% loss of activity after 1 h
65
-
pH 7.0, 30 min, 70% loss of activity
80
-
complete loss of activity after 5 min, activity is retained after 10 min in presence of 1.0 M phosphate buffer
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
dialysis against water causes 50% loss of activity
Citrus sp.
-
immobilized enzyme is very stable and exhibited only a slight decrease in activity upon repeated use
-
immobilized enzyme, the specific activity after the 20th use is the same as the activity prior to the first use
-
unstable after deglycosylation with endoglycosidase H
-
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
25°C, 1 month, more than 80% of the activity is retained
-
25°C, pH 7, unbuffered solution, 3 weeks, little or no loss of activity
-
4°C, stable for 1 year
-
5°C, pH 8.0, immobilized enzyme, 2 years, 4% loss of activity
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expressed in Escherichia coli
-
near homogeneity
-
partial
recombinant enzyme 1.2fold from Escherichia coli strain BL21(DE3) by ammonium sulfate fractionation, gel filtration, dialysis, and ultrafiltration
recombinant enzyme, the expression and purification system has made the industrial production of the enzyme possible
-
recombinant His-tagged enzyme from Escherichia coli by nickel affinity chromatography
-
recombinant N-terminally His-tagged wild-type and selenomethionine-labeled enzymes from Escherichia coli by nickel affinity chromatography, anion exchange chromatography, and gel filtration
recombinant protein purified to 90%
-
strain SHS 0133, FERM BP-2755
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expressed in Acremonium chrysogenum
expressed in Escherichia coli JM109
-
expression in Escherichia coli
-
expression in Escherichia coli; the most efficient expression plasmid is pCAH431, the expression and purification system has made the industrial production of the enzyme possible
-
gene axe, expression of His-tagged enzyme in Escherichia coli
-
gene Cah, subcloning in Escherichia coli strain JM109, expression in Escherichia coli strain BL21(DE3), the enzyme is secreted
gene TM0077, expression of N-terminally His-tagged wild-type enzyme in Escherichia coli and selenomethionine-labeled enzyme in Escherichia coli methionine auxotrophic strain DL41
N-terminally truncated form
-
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
synthesis
Show AA Sequence (329 entries)
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