Information on EC 5.1.1.17 - isopenicillin-N epimerase

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

EC NUMBER
COMMENTARY hide
5.1.1.17
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RECOMMENDED NAME
GeneOntology No.
isopenicillin-N epimerase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
isopenicillin N = penicillin N
show the reaction diagram
forms part of the penicillin biosynthesis pathway; this enzyme contains pyridoxal phosphate, epimerization at C-5 of the 5-amino-5-carboxypentanoyl group to form penicillin N is required to make a substrate for EC 1.14.20.1, deactoxycephalosporin-C synthase, to produce cephalosporins, forms part of the penicillin biosynthesis pathway
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REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
epimerization
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
Biosynthesis of antibiotics
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deacetylcephalosporin C biosynthesis
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Metabolic pathways
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Penicillin and cephalosporin biosynthesis
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SYSTEMATIC NAME
IUBMB Comments
penicillin-N 5-amino-5-carboxypentanoyl-epimerase
This enzyme contains pyridoxal phosphate. Epimerization at C-5 of the 5-amino-5-carboxypentanoyl group to form penicillin N is required to make a substrate for EC 1.14.20.1, deactoxycephalosporin-C synthase, to produce cephalosporins. Forms part of the penicillin biosynthesis pathway (for pathway, click here).
CAS REGISTRY NUMBER
COMMENTARY hide
88201-43-8
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
strain C10 (ATCC 48272) and transformant strains TMCD2, TMCD26, TMCD32, TMCD39, TMCD53, TMCD242, TMCD474
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Manually annotated by BRENDA team
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Manually annotated by BRENDA team
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
deacetoxycephalosporin C
?
show the reaction diagram
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less than 1% of activity with penicillin N
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?
isopenicillin
penicillin N
show the reaction diagram
isopenicillin N
penicillin N
show the reaction diagram
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
isopenicillin N
penicillin N
show the reaction diagram
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
pyridoxal 5'-phosphate
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
4-chloromercuribenzoate
ammonium sulfate
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100 mM, 47% inhibition
Cu2+
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1 mM, complete inhibition
D-cycloserine
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1 mM, 23% inhibition
FAD
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0.5 mM, 80% inhibition
Fe2+
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0.135 mM, complete inhibition
Fluorescamine
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inactivation at molar ratios higher than 2
Hg2+
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1 mM, complete inhibition
hydroxylamine
I-
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1 mM, 27% inhibition
iodoacetamide
Iproniazid
Isoniazid
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1 mM, 23% inhibition
N-ethylmaleimide
Phenylglyoxal
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1 mM, 33% inhibition
Zn2+
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1 mM, 50% inhibition
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.27 - 0.3
isopenicillin N
0.78
Penicillin N
0.0024
pyridoxal 5'-phosphate
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SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.00009
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activity in cell extracts
3.85
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isopenicillin N epimerization
7.77
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penicillin N epimerization
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
8.3
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isopenicillin N epimerase, standard assay mixture contains 1.4 mM isopenicillin N or penicillin N, 0.2 mM dithiothreitol, 0.1 mM pyridoxal phosphate and 50 mM pyrophosphate
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.5 - 8
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strong decrease in activity above pH 8.0 and below pH 6.5
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
25
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slightly lower activity at 20 and 30°C
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5 - 45
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reduced activity below 20°C or above 30°C
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
47000
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1 * 47000, SDS-PAGE
59000
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gel filtration
60000
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gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
monomer
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4.5 - 9
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492261
5.3 - 10
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stable at 4°C for 24 h
492260
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
20 - 40
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stable for 2 h
45
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rapid denaturation above
60
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complete inactivation after 10 min
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
unstable enzyme
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unstable enzyme, pyridoxal 5'-phosphate stabilizes
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STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-65°C, 1 week, 61% loss of activity
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-70°C, 0.05 mM pyridoxal 5'-phosphate, 17 d, no loss of activity
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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
ammonium sulfate, DE-52, DEAE-Affi-gel blue, Sephadex G-200, calcium phosphate, Mono Q
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ammonium sulfate, Sephadex G-200, DEAE-trisacryl
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by anion exchange chromatography, purified IPN epimerase is able to act in both orientations, leading in vitro to an equimolecular mixture of isopenicillin N and penicillin N
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protamine sulfate, ammonium sulfate, Sephadex G-75, Mono Q
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expression in Penicillium chrysogenum
Penecillium chrysogenum strains are constructed expressing cefD1 (isopenecillin N-CoA synthetase), cefD2 (isopenecillin N-CoA epimerase), cefEF (deacetylcephalosporin-actyltransferase), and cefG (deacetylcephalosporin C synthetase) genes cloned from Acremonium chrysogenum. HPLC analysis of cell extracts show that transformant TA64, TA71, and TA98 accumulate intracellularly deacetylcephalosporin C and, in the last strain (TA98), also cephalosporin C.
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EXPRESSION
ORGANISM
UNIPROT
LITERATURE
4% (v/v) glycerol causes the upregulation of isopenicillin N-CoA epimerase in the exponential phase
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
biotechnology
medicine
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the conversion of isopenicillin N into penicillin N is part of the cephalosporin biosynthesis pathway. Cephalosporin is one of the most potent beta-lactam antibiotics, widely used in the treatment of infectious diseases
synthesis
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