Information on EC 6.3.2.26 - N-(5-amino-5-carboxypentanoyl)-L-cysteinyl-D-valine synthase

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

EC NUMBER
COMMENTARY hide
6.3.2.26
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RECOMMENDED NAME
GeneOntology No.
N-(5-amino-5-carboxypentanoyl)-L-cysteinyl-D-valine synthase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
3 ATP + L-2-aminohexanedioate + L-cysteine + L-valine + H2O = 3 AMP + 3 diphosphate + N-[L-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine
show the reaction diagram
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REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
carboxylic acid amid formation
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-
-
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formation of peptide bond
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peptide bond formation
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-
-
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PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
Biosynthesis of antibiotics
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isopenicillin N biosynthesis
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Metabolic pathways
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Penicillin and cephalosporin biosynthesis
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SYSTEMATIC NAME
IUBMB Comments
L-2-aminohexanedioate:L-cysteine:L-valine ligase (AMP-forming, valine-inverting)
Requires Mg2+. The enzyme contains 4'-phosphopantetheine, which may be involved in the mechanism of the reaction. Forms part of the penicillin biosynthesis pathway (for pathway, click here).
CAS REGISTRY NUMBER
COMMENTARY hide
57219-73-5
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
-
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Manually annotated by BRENDA team
Paecilomyces persicinus
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-
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Manually annotated by BRENDA team
Penicillium chrysogenum Wisconsin 54-1255
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
malfunction
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
3 ATP + L-2-aminohexanedioate + L-cysteine + L-valine + H2O
3 AMP + 3 diphosphate + N-[L-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine
show the reaction diagram
6-oxopiperidine 2-carboxylic acid + L-cysteine + L-valine + ATP
N-(5-amino-5-carboxypentanoyl)-L-cysteinyl-D-valine + AMP + diphosphate
show the reaction diagram
-
-
-
-
?
alpha-aminoadipic acid + L-valine + L-lysine + ?
L-delta-alpha-aminoadipyl-L-cysteinyl-D-valine + ?
show the reaction diagram
-
enzyme is a nonribosomal peptide synthetase (NRPS) able to form ribosome-independent peptide bonds
ACV
-
?
ATP + (R)-4-phosphopantoate + beta-alanine
AMP + diphosphate + (R)-4'-phosphopantothenate
show the reaction diagram
DL-valine + L-O-(methylserine) + L-2-aminohexanedioate + ATP
L-O-(methylserinyl)-D-valine + L-O-(methylserinyl)-D-valine + AMP + diphosphate
show the reaction diagram
-
-
-
-
?
L-2-aminoadipate + allylglycine + L-valine + ATP
L-delta-(aminoadipyl)-L-allylglycinyl-D-valine + AMP + diphosphate
show the reaction diagram
L-2-aminoadipate + L-cystathionine + L-valine + ATP
N-(5-amino-5-carboxypentanoyl)-L-cysteinyl-D-valine + AMP + diphosphate
show the reaction diagram
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-
-
-
?
L-2-aminoadipate + L-cysteine + L-allo-isoleucine + ATP
L-delta-(aminoadipyl)-L-cysteinyl-D-allo-isoleucine + AMP + diphosphate
show the reaction diagram
L-2-aminoadipate + L-cysteine + L-isoleucine + ATP
L-delta-(aminoadipyl)-L-cysteinyl-D-isoleucine + AMP + diphosphate
show the reaction diagram
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-
-
-
?
L-2-aminoadipate + L-cysteine + L-valine + ATP
N-(5-amino-5-carboxypentanoyl)-L-cysteinyl-D-valine + AMP + diphosphate
show the reaction diagram
L-2-aminoadipate + L-homocysteine + L-valine + ATP
N-(5-amino-5-carboxypentanoyl)-L-homocysteinyl-D-valine + AMP + diphosphate
show the reaction diagram
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-
-
-
?
L-2-aminoadipate + vinylglycine + L-valine + ATP
L-delta-(aminoadipyl)-L-vinylglycinyl-D-valine + AMP + diphosphate
show the reaction diagram
L-glutamate + L-cysteine + L-valine + ATP
L-glutamyl-L-cysteinyl-D-valine + AMP + diphosphate
show the reaction diagram
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-
-
-
?
S-carboxymethylcysteine + L-cysteine + L-valine + ATP
L-S-carboxymethylcysteinyl-L-cysteinyl-D-valine + AMP + diphosphate
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
3 ATP + L-2-aminohexanedioate + L-cysteine + L-valine + H2O
3 AMP + 3 diphosphate + N-[L-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine
show the reaction diagram
alpha-aminoadipic acid + L-valine + L-lysine + ?
L-delta-alpha-aminoadipyl-L-cysteinyl-D-valine + ?
show the reaction diagram
-
enzyme is a nonribosomal peptide synthetase (NRPS) able to form ribosome-independent peptide bonds
ACV
-
?
ATP + (R)-4-phosphopantoate + beta-alanine
AMP + diphosphate + (R)-4'-phosphopantothenate
show the reaction diagram
Q5JIZ8
the enzyme is involved in coenzyme A biosynthesis in the archaea
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-
?
additional information
?
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COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Mn2+
-
optimal concentration: 10 mM
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
4-phosphopantoate
substrate inhibition
5,5'-dithiobis-2-nitrobenzoate
bis-delta-(L-alpha-aminoadipyl)-L-cysteinyl-D-valine
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D-glucose
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crude extract is inhibited due to deprivation of ATP via sugar metabolism
diphosphate
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5 mM
dithiothreitol
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glyceraldehyde
glyceraldehyde-3-phosphate
iodoacetamide
N-ethylmaleimide
phosphate
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ACVS activity decreases 30% in the presence of 100 mM phosphate, being also affected by AMP and pyrophosphate, products of ATP hydrolysis
pyridoxal 5'-phosphate
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2 mM
additional information
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1.16
(R)-4-phosphopantoate
pH 6.5, 85C
2.44
ATP
pH 6.5, 85C
0.34
beta-Alanine
pH 6.5, 85C
0.045 - 0.63
L-aminoadipate
0.026 - 0.12
L-cysteine
0.08 - 0.34
L-valine
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0833
ATP
Acremonium chrysogenum
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0.133
cysteine
Acremonium chrysogenum
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under saturating conditions
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.38
beta-alanine, pH 6.5, 85C
14.7
ATP, pH 6.5, 85C
15.6
(R)-4-phosphopantoate, pH 6.5, 85C
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7.8
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in MOPS buffer
8.4
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in Tris buffer
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
8 - 8.5
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optimal pH-range for ACVS
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
65 - 95
activity continues to increase with temperature elevation from 65C up to 95C
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
PDB
SCOP
CATH
ORGANISM
UNIPROT
Thermococcus onnurineus (strain NA1)
Thermococcus onnurineus (strain NA1)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
32000
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subunit of Streptomyces clavuligerus ACVS
220000
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gel filtration
283000
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subunit of Streptomyces clavuligerus ACVS
404100
411500
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calculation from amino acid sequence
414800
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calculation from amino acid sequence
422500
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calculation from DNA-sequence, PAGE and gel filtration
424100
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calculation from amino acid sequence
425000
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gel filtration
430000
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gel filtration
470000
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gradient PAGE
560000
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monomer of Streptomyces clavuligerus ACVS that appears as a 500kDa band in SDS-PAGE
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
heterodimer
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ACVS from Streptomyces clavuligerus 2 subunits of 283 kDa and a subunit of 32 kDa
monomer
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
glycoprotein
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pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7
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very unstable below pH 7
440081
8.5
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very unstable above pH 8.5
440081
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
32
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half of activity after 10 min
34
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the mixture of ATP 5 mM, DDT 3 mM, Mg2+ 5 mM, aminoadipate 5 mM, cysteine 1 mM and valine 5 mM prevents the thermal inactivation, 91% activity after 10 min; the mixture of ATP 5 mM, Mg2+ 5 mM, valine 5 mM prevents the thermal inactivation, 92% activity after 10 min
70
6 h, no decrease in activity
80
half-life: 13.6 h
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
glycerol stabilizes enzyme activity
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ORGANIC SOLVENT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
dithiothreitol
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-80C, about 15% of activity after 10 months
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-80C, no loss in activity after 6 weeks
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-80C, several months
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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
ACVS of Streptomyces clavuligerus is purified 12fold using a combination of 2 successive chromatography steps on MonoQ columns separated by ultrafiltration; the recombinant of Streptomyces clavuligerus ACVS is purified from a Streptomyces lividans transformant 2785fold to near homogeneity by a combination of gel filtration, ultrafiltration, and ion-exchange chromatography
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
ACVS of Streptomyces lactamdurans is produced as a recombinant protein from a Streptomyces lividans transformant, carrying the pcbAB gene
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expression in Escherichia coli
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expression in Hansenula polymorpha, co-expression with Bacillus subtilis sfp gene encoding a phosphopantetheinyl transferase that activated ACVS
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expression in Saccharomyces cerevisiae
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gene pcbAB, expression of plasmid p43gdh-ACVSthio encoding the sequence encoding the ACVS thiesterase gene domain, using the Aspergillus awamori gdh gene promoter plus the pcbAB gene transcriptional terminator, in Penicillium chrysogenum protoplasts
overexpressed in Escherichoa coli
overexpression in Streptomyces lividans
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
biotechnology
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production of beta-lactam antibiotics
medicine
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production of penicillin