Information on EC 2.3.1.164 - isopenicillin-N N-acyltransferase

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The expected taxonomic range for this enzyme is: Trichocomaceae

SplaateEC_Number,Commentary
EC NUMBER
COMMENTARY
2.3.1.164
-
SplaateRecommended_Name,GO_Number
RECOMMENDED NAME
GeneOntology No.
isopenicillin-N N-acyltransferase
-
SplaateReaction,Reaction_id,Commentary,IF(Commentary != '',Organism,'') ,IF(Commentary != '',Literature,'')
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
phenylacetyl-CoA + isopenicillin N + H2O = CoA + penicillin G + L-2-aminohexanedioate
show the reaction diagram
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-
-
-
SplaateReaction_Type,Organism,Commentary,Literature
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
Acylation
-
-
-
-
hydrolysis
-
-
-
-
SplaatePathway,BRENDA_Link,KEGG_Link,MetaCyc_Link,Source_Database
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
penicillin K biosynthesis
-
-
Penicillin and cephalosporin biosynthesis
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-
Metabolic pathways
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Biosynthesis of antibiotics
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SplaateSystematic_Name,Commentary_IUBMB
SYSTEMATIC NAME
IUBMB Comments
acyl-CoA:isopenicillin N N-acyltransferase
Proceeds by a two stage mechanism via 6-aminopenicillanic acid. Different from EC 3.5.1.11, penicillin amidase.
SplaateSynonyms,Organism,Commentary,Literature
SYNONYMS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
6-aminopenicillanate acyltransferase
-
-
-
-
6-aminopenicillanic acid acyltransferase
-
-
-
-
6-aminopenicillinanic acid phenylacetyltransferase
-
-
-
-
6-aminopenicillinanic acylase
-
-
-
-
aatA
-
gene encoding isopenicillin N acyltransferase
aatB
-
AN6775.3, gene encoding a penicillin V producing activity, which, in contrast to aatA, does not encode a peroxisomal targeting signal PTS1. aatB homologues also exist in non-penicillin-producing fungi and is suggested to be a paralogue of aatA.
acyl coA:isopenicillin N-acyltransferase
-
acyl coenzyme A: isopenicillin N acyltransferase
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acyl coenzyme A:6-aminopenicillanic acid acyltransferase
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-
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acyl-CoA:6-aminopenicillanate acyltransferase
-
-
-
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acyl-CoA:6-APA acyltransferase
-
-
-
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acyl-CoA:isopenicillin N acyltransferase
-
-
-
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acyl-coenzyme A: IPN acyltransferase
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acyl-coenzyme A:isopenicillin N acyltransferase
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-
-
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acyl-coenzyme A:isopenicillin N-acyltransferase
-
-
-
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acyltransferase, 6-aminopenicillanate
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-
-
-
IAT
Penicillium chrysogenum DS17690
-
-
-
isopenicillin N acyltransferase
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-
-
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isopenicillin N acyltransferase
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-
isopenicillin N acyltransferase
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-
isopenicillin N acyltransferase
Penicillium chrysogenum DS17690
-
-
-
isopenicillin N:acyl CoA acyltransferase
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isopenicillin N:acyl-CoA: acyltransferase
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-
-
-
isopenicillin-N N-acyltransferase
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SplaateCAS_Registry_Number,Commentary
CAS REGISTRY NUMBER
COMMENTARY
54576-90-8
-
SplaateOrganism, Commentary,Literature, Sequence_Code,Sequence_db,Textmining
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
C103A mutant enzyme which has lost its ability to be processed to the mature heterodimeric protein and thus represents the proenzyme form
-
-
Manually annotated by BRENDA team
expression in Hansenula polymorpha
-
-
Manually annotated by BRENDA team
high yielding strain
-
-
Manually annotated by BRENDA team
penDE genes; SC6140, ATCC 2044, source of native enzyme
-
-
Manually annotated by BRENDA team
Penicillium chrysogenum npe6 lacks isopenicillin N acyltransferase activity
-
-
Manually annotated by BRENDA team
Penicillium chrysogenum DS17690
gene penDE
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-
Manually annotated by BRENDA team
Penicillium chrysogenum WIS 54-1255
Wis 54-1255
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-
Manually annotated by BRENDA team
Penicillium chrysogenum yielding
high yielding strain
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-
Manually annotated by BRENDA team
SplaateGeneral_Information, Organism, Commentary, Literature
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
malfunction
-
penicillin production in high-yielding strains can be further improved by the overexpression of IAT while at very high levels of IAT the precursor 6-aminopenicillic acid accumulates
malfunction
Penicillium chrysogenum DS17690
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penicillin production in high-yielding strains can be further improved by the overexpression of IAT while at very high levels of IAT the precursor 6-aminopenicillic acid accumulates
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metabolism
-
catalyses final step of penicillin biosynthesis
physiological function
-
IAT catalyzes a limiting step in high-yielding penicillin production strains
physiological function
Penicillium chrysogenum DS17690
-
IAT catalyzes a limiting step in high-yielding penicillin production strains
-
metabolism
in the last step of penicillin biosynthesis, acyl-CoA:isopenicillin N acyltransferase catalyzes the conversion of isopenicillin N to penicillin G
additional information
substrate binding structure determined by homology modeling and molecular docking in different snapshots, and refined by molecular dynamic simulations. A site located in the region generated by beta1, beta2 and beta5 strands, which forms part of the central structure of beta-subunit, as the potential binding site of isopenicillin N, which establishes interactions with Cys103, Leu169, Gln172, Asp264, and Arg310, detailed overview
SplaateSubstrates,Products,id,Organism_Substrates,Commentary_Substrates, Literature_Substrates, Commentary_Products, Literature_Products,Reversibility
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
phenoxyacetyl-CoA + isopenicillin N + H2O
CoA + penicillin V + L-2-aminohexanedioate
show the reaction diagram
Penicillium chrysogenum, Penicillium chrysogenum DS17690
-
-
-
?
phenylacetyl-CoA + 6-aminopenicillanic acid + H2O
CoA + benzylpenicillin
show the reaction diagram
-
-
-
?
phenylacetyl-CoA + 6-aminopenicillanic acid + H2O
CoA + benzylpenicillin
show the reaction diagram
-
acyl-CoA:6-aminopenicillanate acyltransferase activity
-
?
phenylacetyl-CoA + 6-aminopenicillanic acid + H2O
CoA + benzylpenicillin
show the reaction diagram
Penicillium chrysogenum WIS 54-1255
-
-
-
?
phenylacetyl-CoA + isopenicillin N + H2O
CoA + benzylpenicillin + L-2-aminohexanedioate
show the reaction diagram
-
acyl-coenzyme A:isopenicillin N acyltransferase activity, S309 is involved in substrate acylation
-
?
phenylacetyl-CoA + isopenicillin N + H2O
CoA + penicillin G + L-2-aminohexanedioate
show the reaction diagram
-
-
-
?
phenylacetyl-CoA + isopenicillin N + H2O
CoA + penicillin G + L-2-aminohexanedioate
show the reaction diagram
-
-
-
?
phenylacetyl-CoA + isopenicillin N + H2O
CoA + penicillin G + L-2-aminohexanedioate
show the reaction diagram
-
-
-
?
phenylacetyl-CoA + isopenicillin N + H2O
CoA + penicillin G + L-2-aminohexanedioate
show the reaction diagram
IAT substitutes the alpha-aminoadipic acid side chain of isopenicillin N by a phenylacetic acid phenolate group from phenylacetyl-CoA
-
?
phenylacetyl-CoA + isopenicillin N + H2O
CoA + penicillin G + L-2-aminohexanedioate
show the reaction diagram
Penicillium chrysogenum DS17690
-
-
-
?
SplaateNatural_Substrates,Natural_Products,id,Organism_Substrates,Commentary_Substrates,Literature_Substrates,Commentary_Products,Literature_Products,Reversibility
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
phenylacetyl-CoA + isopenicillin N + H2O
CoA + benzylpenicillin + L-2-aminohexanedioate
show the reaction diagram
-
-
-
?
phenylacetyl-CoA + isopenicillin N + H2O
CoA + penicillin G + L-2-aminohexanedioate
show the reaction diagram
-
-
-
?
phenylacetyl-CoA + isopenicillin N + H2O
CoA + penicillin G + L-2-aminohexanedioate
show the reaction diagram
-
-
-
?
phenylacetyl-CoA + isopenicillin N + H2O
CoA + penicillin G + L-2-aminohexanedioate
show the reaction diagram
-
-
-
?
phenylacetyl-CoA + isopenicillin N + H2O
CoA + penicillin G + L-2-aminohexanedioate
show the reaction diagram
P15802
-
-
?
phenylacetyl-CoA + isopenicillin N + H2O
CoA + penicillin G + L-2-aminohexanedioate
show the reaction diagram
Penicillium chrysogenum DS17690
-
-
-
?
SplaateCofactor,Organism,Commentary,Literature,Filename
SplaateMetals_Ions,Organism,Commentary, Literature
SplaateInhibitors, Organism, Commentary, Literature,Filename
SplaateActivating_Compound, Organism, Commentary, Literature,Filename
SplaateKM_Value,KM_Value_Maximum, Substrate,Organism, Commentary, Literature, Filename
SplaateTurnover_Number, Turnover_Number_Maximum, Substrate,Organism,Commentary, Literature, Filename
SplaateKCat_KM_Value,KCat_KM_Value_Maximum, Substrate,Organism, Commentary, Literature, Filename
SplaateKI_Value,KI_Value_Maximum, Inhibitor,Organism, Commentary, Literature, Filename
SplaateIC50_Value,IC50_Value_Maximum, Inhibitor,Organism, Commentary, Literature, Filename
SplaateSpecific_Activity, Specific_Activity_Maximum, Organism ,Commentary, Literature
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
additional information
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-
SplaatepH_Optimum, pH_Optimum_Maximum, Organism, Commentary, Literature
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
8
-
activity assay
SplaatepH_Range,pH_Range_Maximum, Organism,Commentary, Literature
SplaateTemperature_Optimum, Temperature_Optimum_Maximum, Organism, Commentary, Literature
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
26
-
activity assay
SplaateTemperature_Range, Temperature_Range_Maximum, Organism, Commentary, Literature
SplaatepI_Value,pI_Value_Maximum, Organism,Commentary, Literature
SplaateSource_Tissue, Organism, Commentary, Literature, Textmining
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SOURCE
additional information
-
the protein level of IAT already saturates at low cluster copy numbers, lack of correlation between the protein and transcript levels of the IAT enzyme
Manually annotated by BRENDA team
additional information
Penicillium chrysogenum DS17690
-
the protein level of IAT already saturates at low cluster copy numbers, lack of correlation between the protein and transcript levels of the IAT enzyme
-
Manually annotated by BRENDA team
SplaateLocalization, Organism, Commentary, id_go, Literature, Textmining
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
-
IAT encoded by the aatB gene, lacking a peroxisomal targeting signal PTS1
Manually annotated by BRENDA team
-
colocalization of isopenicillin N-acyltransferase and phenylacetyl-CoA ligase
Manually annotated by BRENDA team
-
both native and recombinant enzyme
Manually annotated by BRENDA team
-
IAT encoded by the aatA gene, containing a peroxisomal targeting signal PTS1
Manually annotated by BRENDA team
Penicillium chrysogenum AS-P-78
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colocalization of isopenicillin N-acyltransferase and phenylacetyl-CoA ligase
-
Manually annotated by BRENDA team
SplaatePDB,PDB,PDB,Organism,Uniprot_ID
SplaateMolecular_Weight, Molecular_Weight_Maximum, Organism, Commentary, Literature
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
11000
-
processed protein, subunit alpha, determined by SDS-PAGE and Western blot analysis
686918
28000
-
SDS-PAGE
703815
29000
-
processed protein, subunit beta, determined by SDS-PAGE and Western blot analysis
686918
40000
-
unprocessed precursor polypeptide, proenzyme, SDS-PAGE
441302, 441303
40000
-
recominant enzyme SDS-PAGE and Western blot analysis
441306
40000
-
gel filtration
658618
40000
-
isopenicillin N acyltransferase precursor, proacyltransferase, proIAT, determined by SDS-PAGE and Western blot analysis
686918
40000
pro-enzyme
720282
SplaateSubunits, Organism, Commentary, Literature
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
?
-
recombinant expression in Hansenula polymorpha give a 40 kDa pre-protein that is processed into an 11 and 29 kDa subunit, SDS-PAGE
dimer
-
alphabeta, 1 * 29000 + 1 * 11000, SDS-PAGE
dimer
-
1 * 11000 + 1 * 29000, SDS-PAGE
dimer
three-dimensional structure determined by homology modeling and molecular docking in different snapshots, and refined by molecular dynamic simulations, detailed overview. The gene for IAT, penDE, encodes a 357 residue proenzyme with a molecular weight of 40 kDa, which is processed autocatalytically to generate two polypeptides of 102 residues as alpha-subunit and 255 residues as beta-subunit
heterodimer
-
alpha 1 * 11000 + beta 1 * 29000, derived from cleavage of the 40 kDa polypeptide, SDS-PAGE
heterodimer
-
beta 1 * 29000, alpha subunit not detected
heterodimer
-
alpha 1 * 11000 + beta 1 * 29000, wild-type and recombinant enzyme, SDS-PAGE
heterodimer
Penicillium chrysogenum WIS 54-1255
-
beta 1 * 29000, alpha subunit not detected
-
additional information
-
isopenicillin N-acyltransferase and phenylacetyl-CoA ligase which supplies phenylacetic acid to the N-acyltransferase form a peroxisomal functional complex
additional information
Penicillium chrysogenum AS-P-78
-
isopenicillin N-acyltransferase and phenylacetyl-CoA ligase which supplies phenylacetic acid to the N-acyltransferase form a peroxisomal functional complex
-
SplaatePosttranslational_Modification, Organism, Commentary, Literature
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
proteolytic modification
-
the intronless gene is expressed in Escherichia coli and self-processed efficiently into its alpha and beta subunits
proteolytic modification
-
recombinant expression in Hansenula polymorpha give a 40 kDa pre-protein that is processed into an 11 and 29 kDa subunit
proteolytic modification
the proenzyme is processed autocatalytically to generate two polypeptides forming alpha-subunit and beta-subunit
additional information
-
ptoIAT overexpressed in Escherichia coli is not self-processed even when treated with Aspergillus nidulans crude extract
SplaateCommentary, Organism, Literature
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
hanging-drop vapour-diffusion method, X-ray diffraction, monoclinic space group C2, unit-cell parameteres: a = 231.36 A, b = 68.27 A, c = 151.31 A, beta = 129.6° at -173°C
-
SplaatepH_Stability,pH_Stability_Maximum, Organism, Commentary, Literature
SplaateTemperature_Stability,Temperature_Stability_Maximum, Organism, Commentary, Literature
SplaateGeneral_Stability, Organism, Literature
SplaateOrganic_Solvent, Organism, Commentary, Literature
SplaateOxidation_Stability,Organism,Literature
SplaateStorage_Stability, Organism, Literature
SplaateCommentary, Organism, Literature
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
cell extracts of Penicillium chrysogenum are prepared, Escherichia coli soluble extracts are used for in vitro IAT activity measurement
-
recombinant wild-type enzyme with a C-terminal His-tag is purified in a single step using Ni2+ affinity chelate chromatography. A combination of 5 mM DTT + 250 mM NaCl + 5 mM EDTA is effective in preventing aggregation of the enzyme
-
SplaateCommentary, Organism, Literature
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
overexpression in Escherichia coli, not self-processed even when treated with Aspergillus nidulans crude extract
-
expressed in Escherichia coli JM109
-
expression in Escherichia coli
-
gene penDE, lack of correlation between the protein and transcript levels of the IAT enzyme, quantitative real-time PCR expression analysis and gene copy number determination, overexpression of penDE in Penicillium chrysogenum strains DS47274, DS47273, and DS17690, introduction of penDE gene without introns into these strains in a random manner, via the nonhomologous end-joining pathway. Cooverexpression of penDE with gene phl, encoding the phenylacetic acid CoA ligase leading to production of phenoxyacetyl-CoA and penicillin V
-
pPenIAT and pPenIATC103S are used to express the penDE gene in Penicillium chrysogenum, pPBCalphabeta, pDEC103S and palphabeta-DEC103S are used for expression in Escherichia coli
-
the intronless gene is expressed in Escherichia coli and self-processed efficiently into its alpha and beta subunits
-
SplaateCommentary, Organism, Literature
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
transcription factor AnBH1 acting as a repressor of aatA and aatB expression
-
transcription factor AnCF acting as an activator of aatA and aatB expression
-
SplaateEngineering, Organism, Commentary, Literature
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
C103S
-
unprocessed inactive variant of isopenicillin N acyltransferase
S227A
-
produces uncleaved proenzyme lacking enzyme activity
S309A
-
recombinant proenzyme containing S309A is cleaved, however enzyme activity is not observed
additional information
-
disruption of the aatA gene reduces penicillin production to 20%, the remaining activity is attributed to the product of gene aatB; Overexpression of a mutated aatB-PTS1 gene in an aatA-disruption strain, leading to peroxisomal localization of AatB, increased the penicillin titre more than overexpression of the wild-type aatB.
C103S or A or W
-
C103 is specifically required for recombinant proenzyme cleavage
additional information
-
in high-yielding penicillin production strains IAT is the limiting factor and that this limitation can be alleviated by a balanced overproduction of this enzyme. Cooverexpression of penDE with gene phl, encoding the phenylacetic acid CoA ligase leading to production of phenoxyacetyl-CoA and penicillin V
T105V
-
results in a significant inhibition of proenzyme cleavage
additional information
Penicillium chrysogenum DS17690
-
in high-yielding penicillin production strains IAT is the limiting factor and that this limitation can be alleviated by a balanced overproduction of this enzyme. Cooverexpression of penDE with gene phl, encoding the phenylacetic acid CoA ligase leading to production of phenoxyacetyl-CoA and penicillin V
-
SplaateCommentary, Organism, Literature
SplaateApplication,Organism,Commentary,Literature
APPLICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
biotechnology
-
Penicillium chrysogenum npe6 lacking isopenicillin N acyltransferase activity is an excellent host for production of different beta-lactam antibiotics