BRENDA - Enzyme Database show
show all sequences of 6.3.2.26

Motifs in the C-terminal region of the Penicillium chrysogenum ACV synthetase are essential for valine epimerization and processivity of tripeptide formation

Wu, X.; Garcia-Estrada, C.; Vaca, I.; Martin, J.F.; Biochimie 94, 354-364 (2012)

Data extracted from this reference:

Cloned(Commentary)
Commentary
Organism
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
Penicillium chrysogenum
Engineering
Amino acid exchange
Commentary
Organism
additional information
exchange of the conserved motifs 3371EGHGRE3376 (located in the putative epimerase domain) and 3629GWSFG3633 (located in the thioesterase domain) by site-directed-mutagenesis to LGFGLL and GWAFG, respectively, combined with deletion of the whole thioesterase domain (230 amino acids) and the different parts of this domain. The mutant enzymes show lower or null activity compared to the control strain
Penicillium chrysogenum
Metals/Ions
Metals/Ions
Commentary
Organism
Structure
Mg2+
the non-ribosomal multidomain ACV synthetase requires ATP and Mg2+
Penicillium chrysogenum
Natural Substrates/ Products (Substrates)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
3 ATP + L-2-aminohexanedioate + L-cysteine + L-valine + H2O
Penicillium chrysogenum
-
3 AMP + 3 diphosphate + N-[L-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine
-
-
?
3 ATP + L-2-aminohexanedioate + L-cysteine + L-valine + H2O
Penicillium chrysogenum Wisconsin 54-1255
-
3 AMP + 3 diphosphate + N-[L-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine
-
-
?
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Penicillium chrysogenum
B6HLU1
a low penicillin production strain, gene pcbAB, the strain contains a single copy of the penicillin gene cluster
-
Penicillium chrysogenum Wisconsin 54-1255
B6HLU1
a low penicillin production strain, gene pcbAB, the strain contains a single copy of the penicillin gene cluster
-
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
3 ATP + L-2-aminohexanedioate + L-cysteine + L-valine + H2O
-
727108
Penicillium chrysogenum
3 AMP + 3 diphosphate + N-[L-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine
-
-
-
?
3 ATP + L-2-aminohexanedioate + L-cysteine + L-valine + H2O
-
727108
Penicillium chrysogenum Wisconsin 54-1255
3 AMP + 3 diphosphate + N-[L-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine
-
-
-
?
Cofactor
Cofactor
Commentary
Organism
Structure
ATP
the non-ribosomal multidomain ACV synthetase requires ATP and Mg2+
Penicillium chrysogenum
Cloned(Commentary) (protein specific)
Commentary
Organism
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
Penicillium chrysogenum
Cofactor (protein specific)
Cofactor
Commentary
Organism
Structure
ATP
the non-ribosomal multidomain ACV synthetase requires ATP and Mg2+
Penicillium chrysogenum
Engineering (protein specific)
Amino acid exchange
Commentary
Organism
additional information
exchange of the conserved motifs 3371EGHGRE3376 (located in the putative epimerase domain) and 3629GWSFG3633 (located in the thioesterase domain) by site-directed-mutagenesis to LGFGLL and GWAFG, respectively, combined with deletion of the whole thioesterase domain (230 amino acids) and the different parts of this domain. The mutant enzymes show lower or null activity compared to the control strain
Penicillium chrysogenum
Metals/Ions (protein specific)
Metals/Ions
Commentary
Organism
Structure
Mg2+
the non-ribosomal multidomain ACV synthetase requires ATP and Mg2+
Penicillium chrysogenum
Natural Substrates/ Products (Substrates) (protein specific)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
3 ATP + L-2-aminohexanedioate + L-cysteine + L-valine + H2O
Penicillium chrysogenum
-
3 AMP + 3 diphosphate + N-[L-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine
-
-
?
3 ATP + L-2-aminohexanedioate + L-cysteine + L-valine + H2O
Penicillium chrysogenum Wisconsin 54-1255
-
3 AMP + 3 diphosphate + N-[L-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine
-
-
?
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
3 ATP + L-2-aminohexanedioate + L-cysteine + L-valine + H2O
-
727108
Penicillium chrysogenum
3 AMP + 3 diphosphate + N-[L-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine
-
-
-
?
3 ATP + L-2-aminohexanedioate + L-cysteine + L-valine + H2O
-
727108
Penicillium chrysogenum Wisconsin 54-1255
3 AMP + 3 diphosphate + N-[L-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine
-
-
-
?
General Information
General Information
Commentary
Organism
additional information
the ACVS is able to catalyse multiple activities including substrate amino acids adenylation, peptide-bond formation, epimerization and tripeptide release by an integrated thioesterase, since it contains three different modules each of approximately 1000 amino acids. The enzyme contains at least ten catalytic domains. The C-terminal region of ACVS bears the epimerase and the thioesterase domains and may be involved in the epimerization of LLL-ACV to LLD-ACV and in the hydrolysis of the thioester bond. Residues E3371, H3373, R3375 and E3376 belong to the epimerase active centre. Different fragments included in the C-terminal region of the enzyme control thioester hydrolysis. Role of the EGHGRE motif present in the epimerase domain of ACVS, the epimerization domain located in the third module (activating L-valine) contains seven partially conserved motifs E1 to E7, overview. The EGHGRE motif containing residues E3371, H3373, R3375 and E3376 is crucial for the activity of the ACVS, involvement of the GWSFG motif in the ACVS activity
Penicillium chrysogenum
General Information (protein specific)
General Information
Commentary
Organism
additional information
the ACVS is able to catalyse multiple activities including substrate amino acids adenylation, peptide-bond formation, epimerization and tripeptide release by an integrated thioesterase, since it contains three different modules each of approximately 1000 amino acids. The enzyme contains at least ten catalytic domains. The C-terminal region of ACVS bears the epimerase and the thioesterase domains and may be involved in the epimerization of LLL-ACV to LLD-ACV and in the hydrolysis of the thioester bond. Residues E3371, H3373, R3375 and E3376 belong to the epimerase active centre. Different fragments included in the C-terminal region of the enzyme control thioester hydrolysis. Role of the EGHGRE motif present in the epimerase domain of ACVS, the epimerization domain located in the third module (activating L-valine) contains seven partially conserved motifs E1 to E7, overview. The EGHGRE motif containing residues E3371, H3373, R3375 and E3376 is crucial for the activity of the ACVS, involvement of the GWSFG motif in the ACVS activity
Penicillium chrysogenum
Other publictions for EC 6.3.2.26
No.
1st author
Pub Med
title
organims
journal
volume
pages
year
Activating Compound
Application
Cloned(Commentary)
Crystallization (Commentary)
Engineering
General Stability
Inhibitors
KM Value [mM]
Localization
Metals/Ions
Molecular Weight [Da]
Natural Substrates/ Products (Substrates)
Organic Solvent Stability
Organism
Oxidation Stability
Posttranslational Modification
Purification (Commentary)
Reaction
Renatured (Commentary)
Source Tissue
Specific Activity [micromol/min/mg]
Storage Stability
Substrates and Products (Substrate)
Subunits
Temperature Optimum [°C]
Temperature Range [°C]
Temperature Stability [°C]
Turnover Number [1/s]
pH Optimum
pH Range
pH Stability
Cofactor
Ki Value [mM]
pI Value
IC50 Value
Activating Compound (protein specific)
Application (protein specific)
Cloned(Commentary) (protein specific)
Cofactor (protein specific)
Crystallization (Commentary) (protein specific)
Engineering (protein specific)
General Stability (protein specific)
IC50 Value (protein specific)
Inhibitors (protein specific)
Ki Value [mM] (protein specific)
KM Value [mM] (protein specific)
Localization (protein specific)
Metals/Ions (protein specific)
Molecular Weight [Da] (protein specific)
Natural Substrates/ Products (Substrates) (protein specific)
Organic Solvent Stability (protein specific)
Oxidation Stability (protein specific)
Posttranslational Modification (protein specific)
Purification (Commentary) (protein specific)
Renatured (Commentary) (protein specific)
Source Tissue (protein specific)
Specific Activity [micromol/min/mg] (protein specific)
Storage Stability (protein specific)
Substrates and Products (Substrate) (protein specific)
Subunits (protein specific)
Temperature Optimum [°C] (protein specific)
Temperature Range [°C] (protein specific)
Temperature Stability [°C] (protein specific)
Turnover Number [1/s] (protein specific)
pH Optimum (protein specific)
pH Range (protein specific)
pH Stability (protein specific)
pI Value (protein specific)
Expression
General Information
General Information (protein specific)
Expression (protein specific)
KCat/KM [mM/s]
KCat/KM [mM/s] (protein specific)
744456
Suring
Biochemical pathways supporti ...
Folsomia candida
Biol. Open
5
1784-1789
2016
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1
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726783
Fernandez-Aguado
A vacuolar membrane protein af ...
Penicillium chrysogenum, Penicillium chrysogenum Wisconsin 54-1255
Appl. Microbiol. Biotechnol.
97
795-808
2013
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727108
Wu
Motifs in the C-terminal regio ...
Penicillium chrysogenum, Penicillium chrysogenum Wisconsin 54-1255
Biochimie
94
354-364
2012
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727436
Ishibashi
A detailed biochemical charact ...
Thermococcus kodakarensis
Extremophiles
16
819-828
2012
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1
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3
3
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700004
Liras
Enzymology of beta-lactam comp ...
Actinomyces sp.
Methods Enzymol.
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401-429
2009
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1
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705518
Siewers
Heterologous production of non ...
Penicillium chrysogenum
Metab. Eng.
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391-397
2009
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706449
Gidijala
An engineered yeast efficientl ...
Penicillium chrysogenum
PLoS ONE
4
e8317
2009
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1
1
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440074
Kallow
Penicillin biosynthesis: energ ...
Acremonium chrysogenum
Biochemistry
37
5947-5952
1998
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1
1
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2
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440073
Kallow
Penicillin biosynthesis: inter ...
Acremonium chrysogenum
FEBS Lett.
414
74-78
1997
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440080
Shiau
L-delta-(alpha-Aminoadipoyl)-L ...
Acremonium chrysogenum
Biochemistry
36
8798-8806
1997
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440084
Theilgaard
-
Purification and characterizat ...
Penicillium chrysogenum
Biochem. J.
327
185-191
1997
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440086
Byford
The mechanism of ACV synthetas ...
Acremonium chrysogenum, Aspergillus nidulans, Penicillium chrysogenum, Streptomyces clavuligerus
Chem. Rev.
97
2631-2649
1997
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2
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440082
Coque
Overexpression of the Nocardia ...
Amycolatopsis lactamdurans
Eur. J. Biochem.
242
264-270
1996
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440079
Shiau
Molecular mechanism of the mul ...
Acremonium chrysogenum
Biochem. Soc. Trans.
23
629S
1995
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440077
Baldwin
-
Substrate specificity of L-del ...
Acremonium chrysogenum, Streptomyces clavuligerus
Biochem. J.
301
367-372
1994
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11
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440085
Aharonowitz
delta-(L-alpha-Aminoadipyl)-L- ...
Acremonium chrysogenum, Amycolatopsis lactamdurans, Aspergillus nidulans, Lysobacter lactamgenus, Paecilomyces persicinus, Penicillium chrysogenum
Bio/Technology
11
807-810
1993
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440078
Zhang
-
Regulation of ACV synthetase a ...
Acremonium chrysogenum, Streptomyces clavuligerus
Arch. Microbiol.
158
364-369
1992
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440081
Zhang
Biochemical studies on the act ...
Streptomyces clavuligerus
Biochem. J.
283
691-698
1992
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8
3
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440083
Zhang
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In vitro stabilization of ACV ...
Streptomyces clavuligerus
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97-110
1992
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440075
MacCabe
delta-(L-alpha-Aminoadipyl)-L- ...
Aspergillus nidulans
J. Biol. Chem.
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12646-12654
1991
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440076
Banko
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delta-(L-alpha-Aminoadipyl)-L- ...
Acremonium chrysogenum
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2858-2860
1987
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