Information on EC 1.21.3.1 - isopenicillin-N synthase

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

EC NUMBER
COMMENTARY
1.21.3.1
-
RECOMMENDED NAME
GeneOntology No.
isopenicillin-N synthase
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine + O2 = isopenicillin N + 2 H2O
show the reaction diagram
removal of 4 hydrogen atoms to form the 4-membered beta-lactam and the 5-membered thiazolidine ring
-
N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine + O2 = isopenicillin N + 2 H2O
show the reaction diagram
Cys106 is involved in substrate binding, Cys255 is involved in maintaining the protein structure
-
N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine + O2 = isopenicillin N + 2 H2O
show the reaction diagram
active site with conserved jelly-roll motif, cysteine residues are not directly involved in the coordination of the metal ion; structure and mechanism
P05326
N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine + O2 = isopenicillin N + 2 H2O
show the reaction diagram
ligation of substrate to the iron centre; structure and mechanism
-
N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine + O2 = isopenicillin N + 2 H2O
show the reaction diagram
structure and mechanism
-
N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine + O2 = isopenicillin N + 2 H2O
show the reaction diagram
structure and mechanism; structure and mechanism
-
N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine + O2 = isopenicillin N + 2 H2O
show the reaction diagram
structure and mechanism
-
N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine + O2 = isopenicillin N + 2 H2O
show the reaction diagram
catalytic reaction is under steric regulation; structure and mechanism
-
N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine + O2 = isopenicillin N + 2 H2O
show the reaction diagram
radical pathway, analogous to the first step of a radicalbased hydroxylation reaction
-
N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine + O2 = isopenicillin N + 2 H2O
show the reaction diagram
the hydroperoxide-ferrous intermediate, formed by O2-activated H atom abstraction from the substrate, can exploit different reaction pathways and interactions with the substrate govern the final pathway
-
N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine + O2 = isopenicillin N + 2 H2O
show the reaction diagram
isopenicillin-N synthase is the main rate-limiting enzyme for penicillin-G biosynthesis
-
N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine + O2 = isopenicillin N + 2 H2O
show the reaction diagram
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
cyclization
-
-
oxidation
-
-
-
-
oxidative cyclization
-
-
oxidative cyclization
-
-
oxidative cyclization
-
-
oxidative cyclization
-
-
oxidative cyclization
-
;
oxidative cyclization
-
-
oxidative cyclization
-
-
oxidative cyclization
Flavobacterium sp. SC 12.154, Streptomyces microflavus NRRL 3584
-
-
-
redox reaction
-
-
-
-
reduction
-
-
-
-
PATHWAY
KEGG Link
MetaCyc Link
Biosynthesis of secondary metabolites
-
isopenicillin N biosynthesis
-
Metabolic pathways
-
Penicillin and cephalosporin biosynthesis
-
SYSTEMATIC NAME
IUBMB Comments
N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine:oxygen oxidoreductase (cyclizing)
Forms part of the penicillin biosynthesis pathway (for pathway, click here).
SYNONYMS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IPN cyclase
-
-
IPN synthase
-
-
IPN synthase
-
ATP is not required, and therefore the enzyme should be named IPN synthase rather than IPN synthetase
IPNS
Flavobacterium sp. SC 12.154
-
-
-
IPNS
Streptomyces microflavus NRRL 3584
-
-
-
isopenicillin N synthase
-
-
-
-
isopenicillin N synthase
-
-
isopenicillin N synthase
-
-
isopenicillin N synthase (cyclase)
-
-
isopenicillin N synthase (cyclase)
-
ATP is not required, and therefore the enzyme should be named IPN synthase rather than IPN synthetase
isopenicillin N synthase (cyclase)
-
-
isopenicillin N synthetase
-
-
-
-
isopenicillin N-synthase
-
-
isopenicillin N-synthase
-
-
isopenicillin N-synthase
Flavobacterium sp. SC 12.154
-
-
-
synthetase, isopenicillin N (9Cl)
-
-
-
-
isopenicillin-N synthase
-
-
additional information
-
isopenicillin N synthase belongs to a class of non-heme ferrous iron dependent oxidoreductases
CAS REGISTRY NUMBER
COMMENTARY
78642-31-6
-
ORGANISM
COMMENTARY
LITERATURE
SEQUENCE CODE
SEQUENCE DB
SOURCE
i.e. Acremonium chrysogenum
-
-
Manually annotated by BRENDA team
strain C10, ATCC 48278
-
-
Manually annotated by BRENDA team
Acremonium chrysogenum C10
strain C10, ATCC 48278
-
-
Manually annotated by BRENDA team
i.e. Nocardia lactamdurans, NRRL 3802 var. JC 1843, cephamycin producing strain, Amy+ mutant
-
-
Manually annotated by BRENDA team
accession number SwissProt
SwissProt
Manually annotated by BRENDA team
inital coordinates for computational calculations are taken from X-ray structure 1BLZ
-
-
Manually annotated by BRENDA team
strain SC 12.154
-
-
Manually annotated by BRENDA team
Flavobacterium sp. SC 12.154
strain SC 12.154
-
-
Manually annotated by BRENDA team
strain NRRL 3584
-
-
Manually annotated by BRENDA team
Streptomyces microflavus NRRL 3584
strain NRRL 3584
-
-
Manually annotated by BRENDA team
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
adipyl-L-cysteinyl-D-valine + ?
N-(4-carboxybutyl)penicillin + 2 H2O
show the reaction diagram
-
-
-
-
?
delta-(L-alpha-aminoadipoyl)-(3R)-methyl-L-cysteine D-alpha hydroxyvaleryl ester + O2
? + H2O
show the reaction diagram
-
-
-
-
?
delta-(L-alpha-aminoadipoyl)-L-cysteinyl-beta-methyl-D-cyclopropylglycine + O2
(2R,8R)-8-([(5S)-5-amino-5-carboxypentanoyl]amino)-3-methylene-9-oxo-6-thia-1-azabicyclo[5.2.0]nonane-2-carboxylic acid + H2O
show the reaction diagram
-
-
-
-
?
delta-(L-alpha-aminoadipoyl)-L-cysteinyl-D-alpha-hydroxyisovaleryl ester + O2
?
show the reaction diagram
-
for this substrate analogue (ACOV) lacking an amide nitrogen IPNS exhibits oxygenase activity
-
-
?
delta-(L-alpha-aminoadipyl)-L-cysteinyl-D-alpha-aminobutyrate + O2
?
show the reaction diagram
-
wild-type and mutants, reaction mechanism
-
-
ir
delta-L-alpha-aminoadipoyl-L-cysteine (1-(S)-carboxy-2-thiomethyl)ethyl ester + O2
? + H2O
show the reaction diagram
-
substrate analogue
-
-
?
delta-L-alpha-aminoadipoyl-L-cysteinyl-D-valine + O2
isopenicillin N + H2O
show the reaction diagram
-
-
-
-
?
delta-L-alpha-aminoadipoyl-L-cysteinyl-D-valine + O2
isopenicillin N + H2O
show the reaction diagram
-
for the native substrate IPNS shows oxidase activity
-
-
?
L-alpha-aminoadipoyl-L-cysteinyl-D-valine + O2
? + H2O
show the reaction diagram
-
-
-
-
?
N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine + O2
isopenicillin N + H2O
show the reaction diagram
P05326
-
-
?
N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine + O2
isopenicillin N + H2O
show the reaction diagram
-
-
-
?
N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine + O2
isopenicillin N + H2O
show the reaction diagram
-
-
-
?
N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine + O2
isopenicillin N + H2O
show the reaction diagram
-
-
-
-
?
N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine + O2
isopenicillin N + H2O
show the reaction diagram
-
-
-
?
N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine + O2
isopenicillin N + H2O
show the reaction diagram
-
-
-
?
N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine + O2
isopenicillin N + H2O
show the reaction diagram
-
-
-
?
N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine + O2
isopenicillin N + H2O
show the reaction diagram
-
-
-
?
N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine + O2
isopenicillin N + H2O
show the reaction diagram
-
-
-
?
N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine + O2
isopenicillin N + H2O
show the reaction diagram
-
-
-
?
N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine + O2
isopenicillin N + H2O
show the reaction diagram
-
-
-
?
N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine + O2
isopenicillin N + H2O
show the reaction diagram
-
-
-
?
N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine + O2
isopenicillin N + H2O
show the reaction diagram
-
-
-
?
N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine + O2
isopenicillin N + H2O
show the reaction diagram
-
-
-
?
N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine + O2
isopenicillin N + H2O
show the reaction diagram
-
-
-
?
N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine + O2
isopenicillin N + H2O
show the reaction diagram
-
-
-
?
N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine + O2
isopenicillin N + H2O
show the reaction diagram
-
-
-
?
N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine + O2
isopenicillin N + H2O
show the reaction diagram
-
-
-
-
?
N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine + O2
isopenicillin N + H2O
show the reaction diagram
-
-
-
?
N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine + O2
isopenicillin N + H2O
show the reaction diagram
-
-
-
?
N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine + O2
isopenicillin N + H2O
show the reaction diagram
P05326
-
-
?
N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine + O2
isopenicillin N + H2O
show the reaction diagram
-
-
-
?
N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine + O2
isopenicillin N + H2O
show the reaction diagram
-
-
-
?
N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine + O2
isopenicillin N + H2O
show the reaction diagram
-
catalytic reaction is under steric regulation
-, product has antibiotic activity
?
N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine + O2
isopenicillin N + H2O
show the reaction diagram
-
common step in the biosynthesis of penicillins, cephalosporins and cephamycins
-
-
?
N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine + O2
isopenicillin N + H2O
show the reaction diagram
-
common step in the biosynthesis of penicillins, cephalosporins and cephamycins
-
?
N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine + O2
isopenicillin N + H2O
show the reaction diagram
-
common step in the biosynthesis of penicillins, cephalosporins and cephamycins
-
-
?
N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine + O2
isopenicillin N + H2O
show the reaction diagram
-
common step in the biosynthesis of penicillins, cephalosporins and cephamycins
-
?
N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine + O2
isopenicillin N + H2O
show the reaction diagram
-
common step in the biosynthesis of penicillins, cephalosporins and cephamycins
-
?
N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine + O2
isopenicillin N + H2O
show the reaction diagram
-
common step in the biosynthesis of penicillins, cephalosporins and cephamycins
-
-
?
N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine + O2
isopenicillin N + H2O
show the reaction diagram
-
common step in the biosynthesis of penicillins, cephalosporins and cephamycins
-
?
N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine + O2
isopenicillin N + H2O
show the reaction diagram
-
common step in the biosynthesis of penicillins, cephalosporins and cephamycins
-
-
?
N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine + O2
isopenicillin N + H2O
show the reaction diagram
P05326
common step in the biosynthesis of penicillins, cephalosporins and cephamycins
product has antibiotic activity
?
N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine + O2
isopenicillin N + H2O
show the reaction diagram
-
common step in the biosynthesis of penicillins, cephalosporins and cephamycins
product has antibiotic activity
?
N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine + O2
isopenicillin N + H2O
show the reaction diagram
-
common step in the biosynthesis of penicillins, cephalosporins and cephamycins
product has antibiotic activity
?
N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine + O2
isopenicillin N + H2O
show the reaction diagram
-
common step in the biosynthesis of penicillins, cephalosporins and cephamycins
product has antibiotic activity
?
N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine + O2
isopenicillin N + H2O
show the reaction diagram
-
common step in the biosynthesis of penicillins, cephalosporins and cephamycins
product has antibiotic activity
?
N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine + O2
isopenicillin N + H2O
show the reaction diagram
-
common step in the biosynthesis of penicillins, cephalosporins and cephamycins
product has antibiotic activity
?
N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine + O2
isopenicillin N + H2O
show the reaction diagram
-
common step in the biosynthesis of penicillins, cephalosporins and cephamycins
product has antibiotic activity
?
N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine + O2
isopenicillin N + H2O
show the reaction diagram
-
common step in the biosynthesis of penicillins, cephalosporins and cephamycins
product has antibiotic activity
?
N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine + O2
isopenicillin N + H2O
show the reaction diagram
-
common step in the biosynthesis of penicillins, cephalosporins and cephamycins
product has antibiotic activity
?
N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine + O2
isopenicillin N + H2O
show the reaction diagram
P05326
common step in the biosynthesis of penicillins, cephalosporins and cephamycins
product has antibiotic activity
?
N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine + O2
isopenicillin N + H2O
show the reaction diagram
-
common step in the biosynthesis of penicillins, cephalosporins and cephamycins
product has antibiotic activity
?
N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine + O2
isopenicillin N + H2O
show the reaction diagram
-
modifications of the L-cysteine residue in the second position of the enzyme result in tripeptides that are unable to serve as substrates
-
?
N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine + O2
isopenicillin N + H2O
show the reaction diagram
-
evaluation of culture conditions for penicillin and cephalosporin C production
product has antibiotic activity
?
N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine + O2
isopenicillin N + H2O
show the reaction diagram
-
key enzyme of the biosynthetic pathway
product has antibiotic activity
?
N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine + O2
isopenicillin N + H2O
show the reaction diagram
-
involved in biosynthesis of cephalosporin C
-
-
?
N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine + O2
isopenicillin N + H2O
show the reaction diagram
Flavobacterium sp. SC 12.154
-
-
-
?
N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine + O2
isopenicillin N + H2O
show the reaction diagram
Flavobacterium sp. SC 12.154
-
common step in the biosynthesis of penicillins, cephalosporins and cephamycins
product has antibiotic activity
?
N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine + O2
isopenicillin N + H2O
show the reaction diagram
Streptomyces microflavus NRRL 3584
-
-
-
?
N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine + O2
isopenicillin N + H2O
show the reaction diagram
Streptomyces microflavus NRRL 3584
-
common step in the biosynthesis of penicillins, cephalosporins and cephamycins
-
-
?
N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine + O2
isopenicillin N + H2O
show the reaction diagram
Acremonium chrysogenum C10
-
-, involved in biosynthesis of cephalosporin C
-
-
?
N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine + O2
isopenicillin + H2O
show the reaction diagram
-
-, the enzyme catalyzes the four electron oxidative double ring closure of its substrate
-
-
?
N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine + O2
isopenicillin N + 2 H2O
show the reaction diagram
-
IPNSs are nonheme iron-dependent oxygenases that catalyze cyclization
-
-
?
N6-[(1R,2S)-1-([[(1R)-1-carboxy-2-methylpropyl]oxy]carbonyl)-2-sulfanylpropyl]-6-oxo-L-lysine + O2
? + H2O
show the reaction diagram
-
substrate analogue
-
-
?
phenylacetyl-L-cysteinyl-D-valine + O2
penicillin G + 2 H2O
show the reaction diagram
-
-
-
-
?
L-alpha-aminoadipoyl-L-cysteinyl-D-valine + O2
isopenicillin N + H2O
show the reaction diagram
-
-
-
-
?
additional information
?
-
-
delta-(L-alpha-aminoadipoyl)-L-cysteinyl-D-alpha-aminobutyrate as substrate is converted to 3 different products: an alpha- and a beta-methyl-penam, and a cepham
-
-
-
additional information
?
-
-
in Penicillium chrysogenum, the enzymes involved in penicillin production are compartmentalized in the cytosol and in microbodies
-
-
-
additional information
?
-
-
iron-mediated conversion of metal-bound thiolate to sulfenate in crystallographic studies
-
-
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
adipyl-L-cysteinyl-D-valine + ?
N-(4-carboxybutyl)penicillin + 2 H2O
show the reaction diagram
-
-
-
-
?
delta-L-alpha-aminoadipoyl-L-cysteinyl-D-valine + O2
isopenicillin N + H2O
show the reaction diagram
-
-
-
-
?
L-alpha-aminoadipoyl-L-cysteinyl-D-valine + O2
? + H2O
show the reaction diagram
-
-
-
-
?
N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine + O2
isopenicillin N + H2O
show the reaction diagram
-
-
-
-
?
N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine + O2
isopenicillin N + H2O
show the reaction diagram
-
-
-
-
?
N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine + O2
isopenicillin N + H2O
show the reaction diagram
-
catalytic reaction is under steric regulation
product has antibiotic activity
?
N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine + O2
isopenicillin N + H2O
show the reaction diagram
-
common step in the biosynthesis of penicillins, cephalosporins and cephamycins
-
-
?
N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine + O2
isopenicillin N + H2O
show the reaction diagram
-
common step in the biosynthesis of penicillins, cephalosporins and cephamycins
-
?
N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine + O2
isopenicillin N + H2O
show the reaction diagram
-
common step in the biosynthesis of penicillins, cephalosporins and cephamycins
-
-
?
N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine + O2
isopenicillin N + H2O
show the reaction diagram
-
common step in the biosynthesis of penicillins, cephalosporins and cephamycins
-
?
N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine + O2
isopenicillin N + H2O
show the reaction diagram
-
common step in the biosynthesis of penicillins, cephalosporins and cephamycins
-
?
N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine + O2
isopenicillin N + H2O
show the reaction diagram
-
common step in the biosynthesis of penicillins, cephalosporins and cephamycins
-
-
?
N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine + O2
isopenicillin N + H2O
show the reaction diagram
-
common step in the biosynthesis of penicillins, cephalosporins and cephamycins
-
?
N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine + O2
isopenicillin N + H2O
show the reaction diagram
-
common step in the biosynthesis of penicillins, cephalosporins and cephamycins
-
-
?
N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine + O2
isopenicillin N + H2O
show the reaction diagram
P05326
common step in the biosynthesis of penicillins, cephalosporins and cephamycins
product has antibiotic activity
?
N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine + O2
isopenicillin N + H2O
show the reaction diagram
-
common step in the biosynthesis of penicillins, cephalosporins and cephamycins
product has antibiotic activity
?
N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine + O2
isopenicillin N + H2O
show the reaction diagram
-
common step in the biosynthesis of penicillins, cephalosporins and cephamycins
product has antibiotic activity
?
N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine + O2
isopenicillin N + H2O
show the reaction diagram
-
common step in the biosynthesis of penicillins, cephalosporins and cephamycins
product has antibiotic activity
?
N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine + O2
isopenicillin N + H2O
show the reaction diagram
-
common step in the biosynthesis of penicillins, cephalosporins and cephamycins
product has antibiotic activity
?
N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine + O2
isopenicillin N + H2O
show the reaction diagram
-
common step in the biosynthesis of penicillins, cephalosporins and cephamycins
product has antibiotic activity
?
N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine + O2
isopenicillin N + H2O
show the reaction diagram
-
common step in the biosynthesis of penicillins, cephalosporins and cephamycins
product has antibiotic activity
?
N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine + O2
isopenicillin N + H2O
show the reaction diagram
-
common step in the biosynthesis of penicillins, cephalosporins and cephamycins
product has antibiotic activity
?
N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine + O2
isopenicillin N + H2O
show the reaction diagram
-
common step in the biosynthesis of penicillins, cephalosporins and cephamycins
product has antibiotic activity
?
N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine + O2
isopenicillin N + H2O
show the reaction diagram
P05326
common step in the biosynthesis of penicillins, cephalosporins and cephamycins
product has antibiotic activity
?
N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine + O2
isopenicillin N + H2O
show the reaction diagram
-
common step in the biosynthesis of penicillins, cephalosporins and cephamycins
product has antibiotic activity
?
N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine + O2
isopenicillin N + H2O
show the reaction diagram
-
modifications of the L-cysteine residue in the second position of the enzyme result in tripeptides that are unable to serve as substrates
-
?
N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine + O2
isopenicillin N + H2O
show the reaction diagram
-
evaluation of culture conditions for penicillin and cephalosporin C production
product has antibiotic activity
?
N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine + O2
isopenicillin N + H2O
show the reaction diagram
-
key enzyme of the biosynthetic pathway
product has antibiotic activity
?
N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine + O2
isopenicillin N + H2O
show the reaction diagram
-
involved in biosynthesis of cephalosporin C
-
-
?
N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine + O2
isopenicillin + H2O
show the reaction diagram
-
-
-
-
?
N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine + O2
isopenicillin N + 2 H2O
show the reaction diagram
-
IPNSs are nonheme iron-dependent oxygenases that catalyze cyclization
-
-
?
N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine + O2
isopenicillin N + H2O
show the reaction diagram
Flavobacterium sp. SC 12.154
-
common step in the biosynthesis of penicillins, cephalosporins and cephamycins
product has antibiotic activity
?
N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine + O2
isopenicillin N + H2O
show the reaction diagram
Streptomyces microflavus NRRL 3584
-
common step in the biosynthesis of penicillins, cephalosporins and cephamycins
-
-
?
N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine + O2
isopenicillin N + H2O
show the reaction diagram
Acremonium chrysogenum C10
-
involved in biosynthesis of cephalosporin C
-
-
?
phenylacetyl-L-cysteinyl-D-valine + O2
penicillin G + 2 H2O
show the reaction diagram
-
-
-
-
?
L-alpha-aminoadipoyl-L-cysteinyl-D-valine + O2
isopenicillin N + H2O
show the reaction diagram
-
-
-
-
?
additional information
?
-
-
in Penicillium chrysogenum, the enzymes involved in penicillin production are compartmentalized in the cytosol and in microbodies
-
-
-
METALS and IONS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
Fe2+
-
stimulates
Fe2+
-
low concentration; stimulates
Fe2+
-
His216 and His 272 are ligands to bind the non-heme iron in the active site; required
Fe2+
-
required
Fe2+
-
required; structure of the Fe2+ active site and endogenous ligands
Fe2+
-
required; required; structure of the Fe2+ active site and endogenous ligands; structure of the Fe2+ active site and endogenous ligands
Fe2+
-
iron-binding motif; required; structure of the Fe2+ active site and endogenous ligands
Fe2+
-
iron-binding motif; structure of the Fe2+ active site and endogenous ligands
Fe2+
-
IPNS is a unique mononuclear non-heme Fe enzyme, the enzyme contains [FeNO]7/[FeO2]8 complexes, combination of spectroscopic techniques including EPR, absorbance, circular dichroism, magnetic CD, and variable temperature, variable-field MCD studies of thiolate bonding and the cofactor electronic structure, computational methods and modeling, overview
Fe2+
-
IPNS is iron-dependent
INHIBITORS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
bis[alpha-aminoadipic(-Cys-D-chloroalanine)]
-
-
-
bis[alpha-aminoadipic(-Cys-D-hexafluorovaline)]
-
-
-
bis[alpha-aminoadipic(-Cys-D-Phe)]
-
-
-
bis[alpha-aminoadipic(-Cys-D-Trp)]
-
-
-
bis[alpha-aminoadipic(-Cys-D-Tyr)]
-
-
-
bis[alpha-aminoadipic(-Cys-DL-hexafluorovaline)]
-
-
-
bis[alpha-aminoadipic(-Cys-hexafluorovaline)]
-
-
-
Co2+
-
moderate inhibition
Co2+
-
inhibition of IPNS activity
Cu2+
-
slight inhibition
cystamine
-
100% inhibition at 0.5 mM
cysteamine
-
43% inhibition at 1.5 mM
cysteine
-
60% inhibition at 1 mM
cystine
-
81% inhibition at 1.5 mM
D-glucose 6-phosphate
-
strong inhibition
delta-(L-alpha-aminoadipoyl)-L-cysteinyl-D-cyclopropylglycine
-
-
glutathione
-
slight inhibition
GSH
-
70% inhibition at 1 mM
Mn2+
-
moderate inhibition
Mn2+
-
moderate inhibition
N-ethylmaleimide
-
only Cys106 can be modified, partly blocked by substrate analogue inhibitors
N-ethylmaleimide
-
wild-type and mutants, binds to Cys104 in the active site
N-ethylmaleimide
-
thiol-specific inhibitor of IPNS
N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine
-
substrate inhibition above 5 mM
NH4+
-
inhibition of enzyme formation in vivo, no inhibitory effect in vitro
Ni2+
-
moderate inhibition
pyruvate
-
slight inhibition
Triton X-100
-
inhibits at concentration of 0.5%
Zn2+
-
moderate inhibition
Zn2+
-
moderate inhibition
Zn2+
-
inhibition of IPNS activity
Mn2+
-
inhibition of IPNS activity
additional information
-
-
-
additional information
-
anions F-, I-, Br-, Cl-, NO3-, H2PO4-, (AsO3)3-, (SO4)2- do not affect the activity; cations Na+, K+, Mg2+, Ca2+, Fe3+ do not affect the activity
-
additional information
-
-
-
additional information
-
ATP, NADPH and FAD: no effect; cations Na+, K+, Mg2+, Ca2+, Fe3+ do not affect the activity
-
additional information
-
conversion of the D-valine residue in third position of the enzyme to an aromatic amino acid or to a highly electronegative residue such as trifluorovaline results in elimination of substrate activity and creation of an inhibitor
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
ascorbate
-
reducing agent stimulates IPNS activity
ascorbic acid
-
required
ascorbic acid
-
great stimulation
ascorbic acid
-
required
ascorbic acid
-
5fold stimulation
dithioerythritol
-
absolutely required, highly stimulating
dithiothreitol
-
required, stimulating
dithiothreitol
-
-
DTT
-
reducing agent stimulates IPNS activity
O2
-
required; required
polyethylene glycol 1500
-
20% stimulation up to concentration of 5%
-
Triton X-100
-
leads to 50% stimulation at concentration of 0.01%
KM VALUE [mM]
KM VALUE [mM] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.08
-
delta-(L-alpha-aminoadipyl)-L-cysteinyl-D-valine
-
-
0.12
-
N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine
-
-
0.18
-
N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine
-
-
3.6
-
phenylacetyl-L-cysteinyl-D-valine
-
-
0.18
-
delta-(L-alpha-aminoadipyl)-L-cysteinyl-D-valine
-
-
additional information
-
N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine
-
Km values for ACV exhibit for different cyclases are on the order of 0.2-0.3 mM
TURNOVER NUMBER [1/s]
TURNOVER NUMBER MAXIMUM[1/s]
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
4.1
-
N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine
-
-
Ki VALUE [mM]
Ki VALUE [mM] Maximum
INHIBITOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.9
-
N-ethylmaleimide
-
about
SPECIFIC ACTIVITY [µmol/min/mg]
SPECIFIC ACTIVITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
0.0000000125
-
-
mutant C104S, recombinant, partially purified
0.000000093
-
-
purified recombinant enzyme
0.000000177
-
-
mutant C251S, recombinant, partially purified
0.000000257
-
-
mutant C142S, recombinant, partially purified
0.000000276
-
-
mutant C37S, recombinant, partially purified
0.000000338
-
-
wild-type, recombinant, partially purified
0.00000224
-
-
mutant H49L
0.0000027
-
-
wild-type
0.00000664
-
-
purified recombinant enzyme
0.0000103
-
-
mutant H126L
0.0000112
-
-
mutant H137L
0.0000116
-
-
mutant H116L
0.0000127
-
-
mutant H64L
0.0000136
-
-
wild-type
0.00129
-
-
about, purified enzyme
additional information
-
-
-
additional information
-
-
development of spectrophotometric assay
pH OPTIMUM
pH MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
7
-
-
assay at
pH RANGE
pH RANGE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
6
9
-
30% activity remaining above pH 9.0, highly reduced activity below pH 6.0
TEMPERATURE OPTIMUM
TEMPERATURE OPTIMUM MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
26
-
-
assay at
27
-
-
assay at
TEMPERATURE RANGE
TEMPERATURE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
SOURCE TISSUE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
SOURCE
LOCALIZATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
Flavobacterium sp. SC 12.154
-
-
-
Manually annotated by BRENDA team
-
inactive associated form
Manually annotated by BRENDA team
Flavobacterium sp. SC 12.154
-
inactive associated form
-
Manually annotated by BRENDA team
PDB
SCOP
CATH
ORGANISM
Emericella nidulans (strain FGSC A4 / ATCC 38163 / CBS 112.46 / NRRL 194 / M139)
Emericella nidulans (strain FGSC A4 / ATCC 38163 / CBS 112.46 / NRRL 194 / M139)
Emericella nidulans (strain FGSC A4 / ATCC 38163 / CBS 112.46 / NRRL 194 / M139)
Emericella nidulans (strain FGSC A4 / ATCC 38163 / CBS 112.46 / NRRL 194 / M139)
Emericella nidulans (strain FGSC A4 / ATCC 38163 / CBS 112.46 / NRRL 194 / M139)
Emericella nidulans (strain FGSC A4 / ATCC 38163 / CBS 112.46 / NRRL 194 / M139)
Emericella nidulans (strain FGSC A4 / ATCC 38163 / CBS 112.46 / NRRL 194 / M139)
Emericella nidulans (strain FGSC A4 / ATCC 38163 / CBS 112.46 / NRRL 194 / M139)
Emericella nidulans (strain FGSC A4 / ATCC 38163 / CBS 112.46 / NRRL 194 / M139)
Emericella nidulans (strain FGSC A4 / ATCC 38163 / CBS 112.46 / NRRL 194 / M139)
Emericella nidulans (strain FGSC A4 / ATCC 38163 / CBS 112.46 / NRRL 194 / M139)
Emericella nidulans (strain FGSC A4 / ATCC 38163 / CBS 112.46 / NRRL 194 / M139)
Emericella nidulans (strain FGSC A4 / ATCC 38163 / CBS 112.46 / NRRL 194 / M139)
Emericella nidulans (strain FGSC A4 / ATCC 38163 / CBS 112.46 / NRRL 194 / M139)
Emericella nidulans (strain FGSC A4 / ATCC 38163 / CBS 112.46 / NRRL 194 / M139)
Emericella nidulans (strain FGSC A4 / ATCC 38163 / CBS 112.46 / NRRL 194 / M139)
Emericella nidulans (strain FGSC A4 / ATCC 38163 / CBS 112.46 / NRRL 194 / M139)
Emericella nidulans (strain FGSC A4 / ATCC 38163 / CBS 112.46 / NRRL 194 / M139)
Emericella nidulans (strain FGSC A4 / ATCC 38163 / CBS 112.46 / NRRL 194 / M139)
Emericella nidulans (strain FGSC A4 / ATCC 38163 / CBS 112.46 / NRRL 194 / M139)
Emericella nidulans (strain FGSC A4 / ATCC 38163 / CBS 112.46 / NRRL 194 / M139)
Emericella nidulans (strain FGSC A4 / ATCC 38163 / CBS 112.46 / NRRL 194 / M139)
Emericella nidulans (strain FGSC A4 / ATCC 38163 / CBS 112.46 / NRRL 194 / M139)
Emericella nidulans (strain FGSC A4 / ATCC 38163 / CBS 112.46 / NRRL 194 / M139)
Emericella nidulans (strain FGSC A4 / ATCC 38163 / CBS 112.46 / NRRL 194 / M139)
Emericella nidulans (strain FGSC A4 / ATCC 38163 / CBS 112.46 / NRRL 194 / M139)
Emericella nidulans (strain FGSC A4 / ATCC 38163 / CBS 112.46 / NRRL 194 / M139)
Emericella nidulans (strain FGSC A4 / ATCC 38163 / CBS 112.46 / NRRL 194 / M139)
Emericella nidulans (strain FGSC A4 / ATCC 38163 / CBS 112.46 / NRRL 194 / M139)
Emericella nidulans (strain FGSC A4 / ATCC 38163 / CBS 112.46 / NRRL 194 / M139)
Emericella nidulans (strain FGSC A4 / ATCC 38163 / CBS 112.46 / NRRL 194 / M139)
Emericella nidulans (strain FGSC A4 / ATCC 38163 / CBS 112.46 / NRRL 194 / M139)
Emericella nidulans (strain FGSC A4 / ATCC 38163 / CBS 112.46 / NRRL 194 / M139)
Emericella nidulans (strain FGSC A4 / ATCC 38163 / CBS 112.46 / NRRL 194 / M139)
Emericella nidulans (strain FGSC A4 / ATCC 38163 / CBS 112.46 / NRRL 194 / M139)
Emericella nidulans (strain FGSC A4 / ATCC 38163 / CBS 112.46 / NRRL 194 / M139)
Emericella nidulans (strain FGSC A4 / ATCC 38163 / CBS 112.46 / NRRL 194 / M139)
Emericella nidulans (strain FGSC A4 / ATCC 38163 / CBS 112.46 / NRRL 194 / M139)
Emericella nidulans (strain FGSC A4 / ATCC 38163 / CBS 112.46 / NRRL 194 / M139)
MOLECULAR WEIGHT
MOLECULAR WEIGHT MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
26500
-
-
gel filtration
36500
38000
-
gel filtration and SDS-PAGE
SUBUNITS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
?
-
x * 38000, wild-type and mutants, SDS-PAGE
?
-
x * 37800, DNA sequence determination
?
Streptomyces microflavus NRRL 3584
-
x * 37800, DNA sequence determination
-
monomer
-
1 * 26500, SDS-PAGE
monomer
-
1 * 36500-38000, SDS-PAGE and gel filtration
monomer
Flavobacterium sp. SC 12.154
-
1 * 36500-38000, SDS-PAGE and gel filtration
-
Crystallization/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
crystal structure of the enzyme reveals that the active site of IPNS is buried in a characteristic jelly-roll motif that has been found in other oxygenases
-
enzyme complexed with manganese instead of iron in the active site, more stable; structure analysis
P05326
in complex with substrate analogue delta-(L-alpha-aminoadipoyl)-(3R)-methyl-L-cysteine D-alpha hydroxyvaleryl ester, crystallization with anaerobic conditions and exposure of crystals to oxygen giving a hydroxymethyl/ene product. Discussion of steric and electronic effects around the valinyl isopropyl side chain of the enzymes active side
-
in complex with tripeptyl analogues delta-(L-alpha-aminoadipoyl)-L-cysteinyl-beta-methyl-D-cyclopropylglycine and delta-(L-alpha-aminoadipoyl)-L-cysteinyl-D-cyclopropylglycine, crystallization in presence of Fe-(II) under anaerobic conditions
-
in complex with truncated substrate analogues delta-(L-alpha-aminoadipoyl)-L-cysteinyl-glycine and delta-(L-alpha-aminoadipoyl)-L-cysteinyl-D-alanine in presence of Fe(II) and presence and absence of nitric oxide. C-terminal carboxylate of substrate is oriented toward the active site iron atom
-
structure analysis
-
the crystal structures of isopenicillin N synthase in complex with gamma-(L-alpha-aminoadipoyl)-(3S-methyl)-L-cysteine D-alpha-hydroxyisovaleryl ester and FeII exposed to O2 and unexposed to O2 are solved to resolutions of 2.2 and 1.65 A, respectively
-
the crystal structures of isopenicillin N synthase in complex with L-alpha-aminoadipoyl-L-cysteine (1-(S)-carboxy-2-thiomethyl)ethyl ester and FeII exposed to O2 and unexposed to O2 are determined to 1.4 and 1.8 A resolutions, respectively
-
crystal structure, molecular modeling of the active site structure and the Fe2+-binding motif
-
GENERAL STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
unstable to 20 mM dithiothreitol during storage at -20C, 10% activity remaining
-
OXIDATION STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
enzyme is unstable to oxidizing oxygen species in the reaction solution
-
440327
Purification/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
recombinant from Escherichia coli
-
wild-type and mutants are purified as catalytically latent apoenzymes
-
wild-type and mutants recombinant from Escherichia coli
-
gel filtration
-
recombinant wild-type and mutants from Escherichia coli
-
partially, wild-type and mutants recombinant from Escherichia coli
-
recombinant, solubilized enzyme
-
Cloned/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
expression in Escherichia coli
-
expression of wild-type and mutant in Escherichia coli, evaluation of growth temperature for expression of the soluble mutant and wild-type in the Escherichia coli host BL21 (DE3)
-
expression of wild-type and mutants in Escherichia coli, amino acid sequence comparison
-
overexpression of recombinant enzyme mutants in Cephalosporium acremonium
-
expressed in Escherichia coli
-
expression of wild-type and mutants as maltose-binding fusion proteins in Escherichia coli
-
expression in Escherichia coli
-
functional expression in the cytosol of yeast Hansenula polymorpha, best at 25C or 30C growth temperature, at 37C the recombinant protein is not stable, optimization of culture conditions, overview
-
expression of wild-type and mutants in Escherichia coli
-
overexpression in Escherichia coli K12 strain JM109 in inclusion bodies
-
overexpression in Escherichia coli, DNA sequence analysis
-
EXPRESSION
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
a developed gene regulation model predicts the expression of this rate limiting enzyme based on glucose repression, fast decay of the mRNA encoding for the enzyme as well as the decay of the enzyme itself
-
ENGINEERING
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
C106S
-
site-directed mutagenesis, 63% reduced activity, 14fold increased Km for N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine
C106S/C255S
-
site-directed mutagenesis, 63% reduced activity, 14fold increased Km for N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine
C255S
-
site-directed mutagenesis, 33% reduced activity, 1.4fold increased Km for N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine
H116L
-
site-directed mutagenesis, reduced activity
H126L
-
site-directed mutagenesis, reduced activity
H137L
-
site-directed mutagenesis, reduced activity
H262L
-
site-directed mutagenesis, complete loss of activity
H272L
-
site-directed mutagenesis, complete loss of activity
H49L
-
site-directed mutagenesis, complete loss of activity
H64L
-
site-directed mutagenesis, reduced activity
P285L
-
site-directed mutagenesis, complete loss of activity, increased soluble expression in Escherichia coli host
Q227L
-
site-directed mutagenesis, 55.10% remaining activity compared to wild-type
Q337L
-
site-directed mutagenesis, slightly reduced activity compared to wild-type
Q230L
-
mutant of IPNS shows diminished enzyme activity, residue is highly conserved among dioxygenases and proximal to the active site, is the fourth ligand for the Fe2+ atom
D216E
-
mutant, retains 1% activity
L223A
-
reduced activity with N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine, delta-(L-alpha-aminoadipoyl)-L-cysteinyl-D-alpha-aminobutyrate is a poor substrate
L223I
-
reduced activity with N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine, product spectrum differs from that of the wild-type with delta-(L-alpha-aminoadipoyl)-L-cysteinyl-D-alpha-aminobutyrate as substrate
L223V
-
reduced activity with N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine, product spectrum differs from that of the wild-type with delta-(L-alpha-aminoadipoyl)-L-cysteinyl-D-alpha-aminobutyrate as substrate
C104S
-
single-strand-site-directed mutagenesis, loss of more than 96% activity
C142S
-
single-strand-site-directed mutagenesis, loss of 24% activity
C251S
-
single-strand-site-directed mutagenesis, loss of 47.7% activity
C37S
-
single-strand-site-directed mutagenesis, loss of 18.3% activity
C37S/C142S/C251S
-
triple mutant, conformationally different from wild-type, prepared by recombining fragments of IPNS-encoding gene pcbC from each of the single mutants, loss of more than 90% activity
D214C
-
site-directed mutagenesis, active site mutant, complete loss of activity
D214E
-
site-directed mutagenesis, active site mutant, retains 1% of activity compared to wild-type
D214H
-
site-directed mutagenesis, complete inactive enzyme
D214H
-
site-directed mutagenesis, active site mutant, complete loss of activity
H212D
-
site-directed mutagenesis, active site mutant, complete loss of activity
H212N
-
site-directed mutagenesis, active site mutant, complete loss of activity
H212Q
-
site-directed mutagenesis, active site mutant, complete loss of activity
H268D
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site-directed mutagenesis, active site mutant, complete loss of activity
H268N
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site-directed mutagenesis, active site mutant, complete loss of activity
H268Q
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site-directed mutagenesis, active site mutant, complete loss of activity
additional information
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exchange of Asp214 and His268, and exchange of Asp214 and His 212 by site-directed mutagenesis leads to completely inactive enzymes
Renatured/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
dissolution of the recombinant enzyme from Escherichia coli inclusion bodies with 7 M urea containing 20 mM DTT, 30 mM glycine, pH 10.0, requires vigorous stirring for over 2 hours at room temperature
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APPLICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
biotechnology
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production of beta-lactam antibiotics
synthesis
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direct enzymic synthesis of antibiotics
medicine
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isopenicillin N synthase is a potential target in the design of novel antibiotic compounds
pharmacology
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model system for study of endogenous functions of beta-lactams in bacteria
pharmacology
Flavobacterium sp. SC 12.154
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model system for study of endogenous functions of beta-lactams in bacteria
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