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show all sequences of 2.3.1.221

Interrogation of global active site occupancy of a fungal iterative polyketide synthase reveals strategies for maintaining biosynthetic fidelity

Vagstad, A.L.; Bumpus, S.B.; Belecki, K.; Kelleher, N.L.; Townsend, C.A.; J. Am. Chem. Soc. 134, 6865-6877 (2012)

Data extracted from this reference:

Cloned(Commentary)
Commentary
Organism
expression of N-terminally His6-tagged enzyme domains in Escherichia coli strain BL21(DE3)
Aspergillus parasiticus
Engineering
Amino acid exchange
Commentary
Organism
S1937A
inactive mutant
Aspergillus parasiticus
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
1.46
-
hexanoyl-[acyl-carrier protein]
pH 7.0, 22°C
Aspergillus parasiticus
15.38
-
acetyl-[acyl-carrier protein]
pH 7.0, 22°C
Aspergillus parasiticus
Natural Substrates/ Products (Substrates)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
7 malonyl-CoA + hexanoyl-[acyl-carrier protein]
Aspergillus parasiticus
-
7 CoA + norsolorinic acid anthrone + [acyl-carrier protein] + 7 CO2 + 2 H2O
-
-
?
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Aspergillus parasiticus
-
-
-
Purification (Commentary)
Commentary
Organism
recombinant N-terminally His6-tagged enzyme domains from Escherichia coli strain BL21(DE3)
Aspergillus parasiticus
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
7 malonyl-CoA + acetyl-[acyl-carrier protein]
-
722459
Aspergillus parasiticus
?
-
-
-
?
7 malonyl-CoA + hexanoyl-[acyl-carrier protein]
-
722459
Aspergillus parasiticus
7 CoA + norsolorinic acid anthrone + [acyl-carrier protein] + 7 CO2 + 2 H2O
-
-
-
?
7 malonyl-CoA + hexanoyl-[acyl-carrier protein]
ACP-bound intermediates and major products of the reaction, overview
722459
Aspergillus parasiticus
7 CoA + norsolorinic acid anthrone + [acyl-carrier protein] + 7 CO2 + 2 H2O
-
-
-
?
additional information
processivity of polyketide extension and substrate specificity, overview. The enzyme shows activity against hexanoyl- and acetyl-, but not malonyl-CoA. Rapid loading of extension units onto the carrier domain facilitates efficient chain extension in a manner kinetically favorable to ultimate product formation. Essential roles of the product template and thioesterase domains for cyclization and product release, editing role for the thioesterase domain
722459
Aspergillus parasiticus
?
-
-
-
-
Subunits
Subunits
Commentary
Organism
More
PksA domain structure, from N- to C-terminus including the starter unit: acylcarrier protein transacylase (SAT), beta-ketoacyl synthase (KS), malonyl-CoA: acyl-carrier protein transacylase (MAT), product template (PT), acyl-carrier protein (ACP), and thioesterase/Claisen cyclase (TE/CLC)
Aspergillus parasiticus
Temperature Optimum [°C]
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
22
-
assay at room temperature
Aspergillus parasiticus
Turnover Number [1/s]
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
0.0407
-
acetyl-[acyl-carrier protein]
pH 7.0, 22°C
Aspergillus parasiticus
0.0472
-
hexanoyl-[acyl-carrier protein]
pH 7.0, 22°C
Aspergillus parasiticus
pH Optimum
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7
-
assay at
Aspergillus parasiticus
Cloned(Commentary) (protein specific)
Commentary
Organism
expression of N-terminally His6-tagged enzyme domains in Escherichia coli strain BL21(DE3)
Aspergillus parasiticus
Engineering (protein specific)
Amino acid exchange
Commentary
Organism
S1937A
inactive mutant
Aspergillus parasiticus
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
1.46
-
hexanoyl-[acyl-carrier protein]
pH 7.0, 22°C
Aspergillus parasiticus
15.38
-
acetyl-[acyl-carrier protein]
pH 7.0, 22°C
Aspergillus parasiticus
Natural Substrates/ Products (Substrates) (protein specific)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
7 malonyl-CoA + hexanoyl-[acyl-carrier protein]
Aspergillus parasiticus
-
7 CoA + norsolorinic acid anthrone + [acyl-carrier protein] + 7 CO2 + 2 H2O
-
-
?
Purification (Commentary) (protein specific)
Commentary
Organism
recombinant N-terminally His6-tagged enzyme domains from Escherichia coli strain BL21(DE3)
Aspergillus parasiticus
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
7 malonyl-CoA + acetyl-[acyl-carrier protein]
-
722459
Aspergillus parasiticus
?
-
-
-
?
7 malonyl-CoA + hexanoyl-[acyl-carrier protein]
-
722459
Aspergillus parasiticus
7 CoA + norsolorinic acid anthrone + [acyl-carrier protein] + 7 CO2 + 2 H2O
-
-
-
?
7 malonyl-CoA + hexanoyl-[acyl-carrier protein]
ACP-bound intermediates and major products of the reaction, overview
722459
Aspergillus parasiticus
7 CoA + norsolorinic acid anthrone + [acyl-carrier protein] + 7 CO2 + 2 H2O
-
-
-
?
additional information
processivity of polyketide extension and substrate specificity, overview. The enzyme shows activity against hexanoyl- and acetyl-, but not malonyl-CoA. Rapid loading of extension units onto the carrier domain facilitates efficient chain extension in a manner kinetically favorable to ultimate product formation. Essential roles of the product template and thioesterase domains for cyclization and product release, editing role for the thioesterase domain
722459
Aspergillus parasiticus
?
-
-
-
-
Subunits (protein specific)
Subunits
Commentary
Organism
More
PksA domain structure, from N- to C-terminus including the starter unit: acylcarrier protein transacylase (SAT), beta-ketoacyl synthase (KS), malonyl-CoA: acyl-carrier protein transacylase (MAT), product template (PT), acyl-carrier protein (ACP), and thioesterase/Claisen cyclase (TE/CLC)
Aspergillus parasiticus
Temperature Optimum [°C] (protein specific)
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
22
-
assay at room temperature
Aspergillus parasiticus
Turnover Number [1/s] (protein specific)
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
0.0407
-
acetyl-[acyl-carrier protein]
pH 7.0, 22°C
Aspergillus parasiticus
0.0472
-
hexanoyl-[acyl-carrier protein]
pH 7.0, 22°C
Aspergillus parasiticus
pH Optimum (protein specific)
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7
-
assay at
Aspergillus parasiticus
General Information
General Information
Commentary
Organism
metabolism
the the norsolorinic acid anthrone-producing polyketide synthase, PksA, is involved in the aflatoxin biosynthetic pathway in Aspergillus parasiticus
Aspergillus parasiticus
additional information
the enzyme also has an editing function for the C-terminal thioesterase domain beyond its synthetic role in Claisen/Dieckmann cyclization and product release. Domain architecture and expected enzyme-bound intermediates of PksA, overview
Aspergillus parasiticus
General Information (protein specific)
General Information
Commentary
Organism
metabolism
the the norsolorinic acid anthrone-producing polyketide synthase, PksA, is involved in the aflatoxin biosynthetic pathway in Aspergillus parasiticus
Aspergillus parasiticus
additional information
the enzyme also has an editing function for the C-terminal thioesterase domain beyond its synthetic role in Claisen/Dieckmann cyclization and product release. Domain architecture and expected enzyme-bound intermediates of PksA, overview
Aspergillus parasiticus
KCat/KM [mM/s]
kcat/KM Value [1/mMs-1]
kcat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
0.0026
-
acetyl-[acyl-carrier protein]
pH 7.0, 22°C
Aspergillus parasiticus
0.032
-
hexanoyl-[acyl-carrier protein]
pH 7.0, 22°C
Aspergillus parasiticus
KCat/KM [mM/s] (protein specific)
KCat/KM Value [1/mMs-1]
KCat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
0.0026
-
acetyl-[acyl-carrier protein]
pH 7.0, 22°C
Aspergillus parasiticus
0.032
-
hexanoyl-[acyl-carrier protein]
pH 7.0, 22°C
Aspergillus parasiticus
Other publictions for EC 2.3.1.221
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)
722459
Vagstad
Interrogation of global active ...
Aspergillus parasiticus
J. Am. Chem. Soc.
134
6865-6877
2012
-
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1
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1
-
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2
-
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1
-
2
-
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1
-
-
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-
-
4
1
1
-
-
2
1
-
-
-
-
-
-
-
-
1
-
-
1
-
-
-
-
2
-
-
-
1
-
-
-
1
-
-
-
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4
1
1
-
-
2
1
-
-
-
-
2
2
-
2
2
723638
Korman
Structure and function of an i ...
Aspergillus sp.
Proc. Natl. Acad. Sci. USA
107
6246-6251
2010
-
-
1
1
4
-
-
-
-
-
-
1
-
1
-
-
1
1
-
-
-
-
1
1
1
-
-
-
1
-
-
-
-
-
-
-
-
1
-
1
4
-
-
-
-
-
-
-
-
1
-
-
-
1
-
-
-
-
1
1
1
-
-
-
1
-
-
-
-
2
2
-
-
-
723262
Crawford
Structural basis for biosynthe ...
Aspergillus parasiticus
Nature
461
1139-1143
2009
-
-
1
-
-
-
-
-
-
-
-
1
-
1
-
-
1
1
-
-
-
-
1
1
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
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-
-
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-
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-
-
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1
-
-
-
1
-
-
-
-
1
1
-
-
-
-
-
-
-
-
-
1
1
-
-
-
721052
Crawford
Deconstruction of iterative mu ...
Aspergillus parasiticus
Science
320
243-246
2008
-
-
1
-
-
-
-
-
-
-
-
1
-
1
-
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1
1
-
-
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1
1
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1
-
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-
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1
-
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-
1
-
-
-
-
1
1
-
-
-
-
-
-
-
-
-
2
2
-
-
-
723194
Chang
The Aspergillus parasiticus po ...
Aspergillus parasiticus, Aspergillus parasiticus SRRC 2043 / RHN1
Mol. Gen. Genet.
248
270-277
1995
-
-
1
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1
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2
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1
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1
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1
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1
-
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3
3
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