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

EC NUMBER
COMMENTARY
6.3.2.26
-
RECOMMENDED NAME
GeneOntology No.
N-(5-amino-5-carboxypentanoyl)-L-cysteinyl-D-valine synthase
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
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
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
carboxylic acid amid formation
-
-
-
-
formation of peptide bond
-
-
peptide bond formation
-
-
-
-
PATHWAY
KEGG Link
MetaCyc Link
Biosynthesis of secondary metabolites
-
isopenicillin N biosynthesis
-
Metabolic pathways
-
Penicillin and cephalosporin biosynthesis
-
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).
SYNONYMS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
ACV synthetase
-
-
-
-
ACV synthetase
-
-
ACV synthetase
-
-
DELTA-(alpha-aminoadipyl)cysteinylvaline synthetase
-
-
-
-
delta-(L-alpha-aminoadipoyl)-L-cysteinyl-D-valine synthetase
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-
-
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delta-(L-alpha-aminoadipyl)-L-cysteinyl-D-valine synthetase
-
-
delta-L-(alpha-aminoadipyl)-L-cysteinyl-D-valine synthetase
-
-
-
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L-(alpha-aminoadipyl)-L-cysteinyl-D-valine synthetase
-
-
-
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L-delta-(alpha-aminoadipoyl)-L-cysteinyl-D-valine synthetase
-
-
-
-
L-delta-alpha-aminoadipoyl-L-cysteinyl-D-valine synthetase
-
-
synthetase, delta-(alpha-aminoadipyl)cysteinylvaline
-
-
-
-
CAS REGISTRY NUMBER
COMMENTARY
57219-73-5
-
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
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
-
?
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
-
-
-
-
?
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-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
-
-
-
-
?
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-cysteine + L-valine + ATP
N-(5-amino-5-carboxypentanoyl)-L-cysteinyl-D-valine + AMP + diphosphate
show the reaction diagram
-
-
-
-
-
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-cysteine + L-valine + ATP
N-(5-amino-5-carboxypentanoyl)-L-cysteinyl-D-valine + AMP + diphosphate
show the reaction diagram
-
-
-
-
-
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-cysteine + L-valine + ATP
N-(5-amino-5-carboxypentanoyl)-L-cysteinyl-D-valine + AMP + diphosphate
show the reaction diagram
-
-
-
-
ir
L-2-aminoadipate + L-cysteine + L-valine + ATP
N-(5-amino-5-carboxypentanoyl)-L-cysteinyl-D-valine + AMP + diphosphate
show the reaction diagram
Paecilomyces persicinus, Lysobacter lactamgenus
-
-
-
-
-
L-2-aminoadipate + L-homocysteine + L-valine + ATP
N-(5-amino-5-carboxypentanoyl)-L-homocysteinyl-D-valine + AMP + diphosphate
show the reaction diagram
-
-
-
-
?
L-2-aminoadipate + vinylglycine + L-valine + ATP
L-delta-(aminoadipyl)-L-vinylglycinyl-D-valine + AMP + diphosphate
show the reaction diagram
-
-
-
-
?
S-carboxymethylcysteine + L-cysteine + L-valine + ATP
L-S-carboxymethylcysteinyl-L-cysteinyl-D-valine + AMP + diphosphate
show the reaction diagram
-
-
-
-
?
S-carboxymethylcysteine + L-cysteine + L-valine + ATP
L-S-carboxymethylcysteinyl-L-cysteinyl-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
-
-
-
-
?
additional information
?
-
-
ACVSs are multifunctional enzymes that activate amino acids as aminoacyladenylates, forming a mixed anhydride with the a-phosphate group of ATP and releasing pyrophosphate, pure Streptomyces lactamdurans ACVS is able to activate alpha-aminoadipic acid or its lactam 6-oxopiperideine-2-carboxylic acid, a compound that is easily converted to alpha-aminoadipic acid, but is unable to use piperideine-6-carboxylate or pipecolic acid as substrates, enzyme is also able to use L-cystathionine with the same efficiency as L-cysteine
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-
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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
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
-
?
additional information
?
-
-
ACVSs are multifunctional enzymes that activate amino acids as aminoacyladenylates, forming a mixed anhydride with the a-phosphate group of ATP and releasing pyrophosphate, pure Streptomyces lactamdurans ACVS is able to activate alpha-aminoadipic acid or its lactam 6-oxopiperideine-2-carboxylic acid, a compound that is easily converted to alpha-aminoadipic acid, but is unable to use piperideine-6-carboxylate or pipecolic acid as substrates, enzyme is also able to use L-cystathionine with the same efficiency as L-cysteine
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-
-
COFACTOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
ATP
-
optimal concentration: 1.5-5 mM
ATP
-
optimal concentration: 5 mM
ATP
-
ACVS has a adenylate-containing modules for ATP binding
METALS and IONS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
Mg2+
-
optimal concentration: 10 mM
Mg2+
-
optimal concentration: 10 mM
Mg2+
-
optimal concentration: 20 mM
Mn2+
-
optimal concentration: 10 mM
INHIBITORS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
5,5'-dithiobis-2-nitrobenzoate
-
1 mM
5,5'-dithiobis-2-nitrobenzoate
-
the major effect on ACVS activity is exerted by thiol-blocking agents, which almost completely inhibited the activity at 1 mM concentration, confirming the importance of thiol groups in ACVS activity
ATP
-
at concentrations about 5 mM
bis-delta-(L-alpha-aminoadipyl)-L-cysteinyl-D-valine
-
-
-
D-glucose
-
crude extract is inhibited due to deprivation of ATP via sugar metabolism
glyceraldehyde-3-phosphate
-
reacts with L-cysteinee
iodoacetamide
-
the major effect on ACVS activity is exerted by thiol-blocking agents, which almost completely inhibited the activity at 1 mM concentration, confirming the importance of thiol groups in ACVS activity
N-ethylmaleimide
-
1 mM
N-ethylmaleimide
-
the major effect on ACVS activity is exerted by thiol-blocking agents, which almost completely inhibited the activity at 1 mM concentration, confirming the importance of thiol groups in ACVS activity
phosphate
-
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
-
2 mM
KM VALUE [mM]
KM VALUE [mM] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.045
-
L-aminoadipate
-
-
0.17
-
L-aminoadipate
-
-
0.63
-
L-aminoadipate
-
-
0.026
-
L-cysteine
-
-
0.08
-
L-cysteine
-
-
0.12
-
L-cysteine
-
-
0.34
-
L-valine
-
-
TURNOVER NUMBER [1/s]
TURNOVER NUMBER MAXIMUM[1/s]
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.133
-
cysteine
-
under saturating conditions
SPECIFIC ACTIVITY [µmol/min/mg]
SPECIFIC ACTIVITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
pH OPTIMUM
pH MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
7.5
-
-
assay at
7.8
-
-
in MOPS buffer
8.4
-
-
in Tris buffer
pH RANGE
pH RANGE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
8
8.5
-
optimal pH-range for ACVS
TEMPERATURE OPTIMUM
TEMPERATURE OPTIMUM MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
25
-
-
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
MOLECULAR WEIGHT
MOLECULAR WEIGHT MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
32000
-
-
subunit of Streptomyces clavuligerus ACVS
220000
-
-
gel filtration
283000
-
-
subunit of Streptomyces clavuligerus ACVS
404100
-
-
calculation from amino acid sequence
404100
-
-
pcbAB gene from Nocardia lactamdurans is 10.9 kb long and encodes a 3649 amino acid ACVS
411500
-
-
calculation from amino acid sequence
414800
-
-
calculation from amino acid sequence
422500
-
-
calculation from DNA-sequence, PAGE and gel filtration
424100
-
-
calculation from amino acid sequence
425000
-
-
gel filtration
430000
-
-
gel filtration
470000
-
-
gradient PAGE
560000
-
-
monomer of Streptomyces clavuligerus ACVS that appears as a 500kDa band in SDS-PAGE
SUBUNITS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
heterodimer
-
ACVS from Streptomyces clavuligerus 2 subunits of 283 kDa and a subunit of 32 kDa
monomer
-
1 * 400000, SDS-PAGE
monomer
-
1 * 430000, SDS-PAGE
monomer
-
1 * 470000, SDS-PAGE
monomer
-
1 * 220000, SDS-PAGE
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
glycoprotein
-
-
pH STABILITY
pH STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
7
-
-
very unstable below pH 7
8.5
-
-
very unstable above pH 8.5
TEMPERATURE STABILITY
TEMPERATURE STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
4
-
-
half of activity after 70 h
25
-
-
half of activity after 10 h
25
-
-
half of activity after 8 h
32
-
-
half of activity after 10 min
34
-
-
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
GENERAL STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
glycerol stabilizes enzyme activity
-
ORGANIC SOLVENT
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
dithiothreitol
-
8 mM leads to a 10% higher enzyme activity than at 3 mM
dithiothreitol
-
stability of crude enzyme is increased by dithiothreitol
STORAGE STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
-80°C, about 15% of activity after 10 months
-
-80°C, no loss in activity after 6 weeks
-
-80°C, several months
-
Purification/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
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
-
Cloned/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
ACVS of Streptomyces lactamdurans is produced as a recombinant protein from a Streptomyces lividans transformant, carrying the pcbAB gene
-
overexpression in Streptomyces lividans
-
expression in Escherichia coli
-
expression in Hansenula polymorpha, co-expression with Bacillus subtilis sfp gene encoding a phosphopantetheinyl transferase that activated ACVS
-
expression in Saccharomyces cerevisiae
-
APPLICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
biotechnology
-
production of beta-lactam antibiotics
medicine
-
production of penicillin