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show all sequences of 5.5.1.B6

Crystal structure of the polyketide cyclase AknH with bound substrate and product analogue: implications for catalytic mechanism and product stereoselectivity

Kallio, P.; Sultana, A.; Niemi, J.; Maentsaelae, P.; Schneider, G.; J. Mol. Biol. 357, 210-220 (2006)

Data extracted from this reference:

Cloned(Commentary)
Commentary
Organism
expression in Escherichia coli
Streptomyces galilaeus
Crystallization (Commentary)
Crystallization
Organism
AknH forms a tetramer of 222 symmetry. Two subunits (A-B and C-D, respectively), related by one of the 2-fold symmetry axes, form tight dimer interfaces. The side-chains of Thr128, Gln105 and the main-chain nitrogen of Val92 each interact with oxygen atoms of the polyketide core of the product via a water molecule. Additional enzyme-ligand interactions involve residues Arg120 and Trp122 from the other monomer across the subunit-subunit interface
Streptomyces galilaeus
Engineering
Amino acid exchange
Commentary
Organism
D121A
0.2% of wild-type activity, (R)-configuration at C9 of product
Streptomyces galilaeus
H107A
2% of wild-type activity, (R)-configuration at C9 of product
Streptomyces galilaeus
N51L
exchange to corresponding residues of SnoaL, EC 5.5.1.26, producing product with (S)-configuration. 20% of wild-type activity, about 50% (S)-configuration at C9 of product
Streptomyces galilaeus
Q105A
6% of wild-type activity, (R)-configuration at C9 of product
Streptomyces galilaeus
Y15F
exchange to corresponding residues of SnoaL, EC 5.5.1.26, producing product with (S)-configuration. 45% of wild-type activity, about 20% (S)-configuration at C9 of product
Streptomyces galilaeus
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.002
-
nogalonic acid methyl ester
pH 7.4, temperature not specified in the publication
Streptomyces galilaeus
Natural Substrates/ Products (Substrates)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
aklanonic acid methyl ester
Streptomyces galilaeus
-
?
-
-
?
nogalonic acid methyl ester
Streptomyces galilaeus
-
auraviketone
product has (R)-configuration at C9, ring closure is an intramolecular aldol condensation. The attack of the enol(ate) is on the si face of the C9 carbonyl group, required for the formation of the C9-R stereoisomer, which results in the alcoholate anion pointing away from the carboxyl side-chain of Asp121
-
?
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Streptomyces galilaeus
O52646
-
-
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
aklanonic acid methyl ester
-
732336
Streptomyces galilaeus
?
-
-
-
?
nogalonic acid methyl ester
-
732336
Streptomyces galilaeus
auraviketone
product has (R)-configuration at C9, ring closure is an intramolecular aldol condensation. The attack of the enol(ate) is on the si face of the C9 carbonyl group, required for the formation of the C9-R stereoisomer, which results in the alcoholate anion pointing away from the carboxyl side-chain of Asp121
-
-
?
Turnover Number [1/s]
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
1
-
nogalonic acid methyl ester
pH 7.4, temperature not specified in the publication
Streptomyces galilaeus
pH Optimum
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7.2
-
-
Streptomyces galilaeus
Cloned(Commentary) (protein specific)
Commentary
Organism
expression in Escherichia coli
Streptomyces galilaeus
Crystallization (Commentary) (protein specific)
Crystallization
Organism
AknH forms a tetramer of 222 symmetry. Two subunits (A-B and C-D, respectively), related by one of the 2-fold symmetry axes, form tight dimer interfaces. The side-chains of Thr128, Gln105 and the main-chain nitrogen of Val92 each interact with oxygen atoms of the polyketide core of the product via a water molecule. Additional enzyme-ligand interactions involve residues Arg120 and Trp122 from the other monomer across the subunit-subunit interface
Streptomyces galilaeus
Engineering (protein specific)
Amino acid exchange
Commentary
Organism
D121A
0.2% of wild-type activity, (R)-configuration at C9 of product
Streptomyces galilaeus
H107A
2% of wild-type activity, (R)-configuration at C9 of product
Streptomyces galilaeus
N51L
exchange to corresponding residues of SnoaL, EC 5.5.1.26, producing product with (S)-configuration. 20% of wild-type activity, about 50% (S)-configuration at C9 of product
Streptomyces galilaeus
Q105A
6% of wild-type activity, (R)-configuration at C9 of product
Streptomyces galilaeus
Y15F
exchange to corresponding residues of SnoaL, EC 5.5.1.26, producing product with (S)-configuration. 45% of wild-type activity, about 20% (S)-configuration at C9 of product
Streptomyces galilaeus
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.002
-
nogalonic acid methyl ester
pH 7.4, temperature not specified in the publication
Streptomyces galilaeus
Natural Substrates/ Products (Substrates) (protein specific)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
aklanonic acid methyl ester
Streptomyces galilaeus
-
?
-
-
?
nogalonic acid methyl ester
Streptomyces galilaeus
-
auraviketone
product has (R)-configuration at C9, ring closure is an intramolecular aldol condensation. The attack of the enol(ate) is on the si face of the C9 carbonyl group, required for the formation of the C9-R stereoisomer, which results in the alcoholate anion pointing away from the carboxyl side-chain of Asp121
-
?
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
aklanonic acid methyl ester
-
732336
Streptomyces galilaeus
?
-
-
-
?
nogalonic acid methyl ester
-
732336
Streptomyces galilaeus
auraviketone
product has (R)-configuration at C9, ring closure is an intramolecular aldol condensation. The attack of the enol(ate) is on the si face of the C9 carbonyl group, required for the formation of the C9-R stereoisomer, which results in the alcoholate anion pointing away from the carboxyl side-chain of Asp121
-
-
?
Turnover Number [1/s] (protein specific)
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
1
-
nogalonic acid methyl ester
pH 7.4, temperature not specified in the publication
Streptomyces galilaeus
pH Optimum (protein specific)
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7.2
-
-
Streptomyces galilaeus
Other publictions for EC 5.5.1.B6
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)
732336
Kallio
Crystal structure of the polyk ...
Streptomyces galilaeus
J. Mol. Biol.
357
210-220
2006
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1
1
5
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1
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2
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1
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2
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1
1
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1
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1
5
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-
1
-
-
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2
-
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-
-
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2
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1
1
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