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show all sequences of 1.14.11.B17

Stereospecific synthesis of threo- and erythro-beta-hydroxyglutamic acid during kutzneride biosynthesis

Strieker, M.; Nolan, E.M.; Walsh, C.T.; Marahiel, M.A.; J. Am. Chem. Soc. 131, 13523-13530 (2009)

Data extracted from this reference:

Cloned(Commentary)
Commentary
Organism
overproduced in Escherichia coli as His7-tagged fusions
Kutzneria sp. 744
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
additional information
-
additional information
nonhydrolyzable coenzyme A analogs are developed and used to determine the kinetic parameters for KtzO-catalyzed hydroxylation of glutamic acid bound to the carrier protein. To determine the kinetic parameters of KtzO catalyzed hydroxylation, the problem of the labile thioester bond is circumvented by using synthetic coenzyme A analogs coupled to glutamic acid where the thioester is replaced by a hydrolytically stable amide bond
Kutzneria sp. 744
Metals/Ions
Metals/Ions
Commentary
Organism
Structure
Iron
putative non-heme iron oxygenase, exhibit the conserved HXD/E_H iron(II)-binding motif, in the assay (NH4)2FeSO4 is used as source of the ferrous iron cofactor (0.5 mM)
Kutzneria sp. 744
Molecular Weight [Da]
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
40100
-
x * 40100, SDS-PAGE
Kutzneria sp. 744
Natural Substrates/ Products (Substrates)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
S-(L-glutamyl)-[peptidyl-carrier protein of nonribosomal peptide synthetase KtzH] + 2-oxoglutarate + O2
Kutzneria sp. 744
the enzyme is involved in the biosynthesis of L-threo-3-hydroxy-glutamate, prior to incorporation into kutznerides (antifungal nonribosomal hexadepsipeptides)
S-(threo-3-hydroxy-L-glutamyl)-[peptidyl-carrier-protein of nonribosomal peptide synthetase KtzH] + succinate + CO2
-
-
?
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Kutzneria sp. 744
A8CF77
-
-
Purification (Commentary)
Commentary
Organism
-
Kutzneria sp. 744
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
additional information
able to hydroxylate glutamic acid bound to a noncognate peptidyl-carrier-protein, e.g. the ninth peptidyl-carrier-protein domain of the CDA NRPS assembly line (CDA-T9). Nonhydrolyzable coenzyme A analogs are developed and used to determine the kinetic parameters for KtzO-catalyzed hydroxylation of glutamic acid bound to the carrier protein. To determine the kinetic parameters of KtzO catalyzed hydroxylation, the problem of the labile thioester bond is circumvented by using synthetic coenzyme A analogs coupled to glutamic acid where the thioester is replaced by a hydrolytically stable amide bond
725193
Kutzneria sp. 744
?
-
-
-
-
S-(L-glutamyl)-[peptidyl-carrier protein of nonribosomal peptide synthetase KtzH] + 2-oxoglutarate + O2
the enzyme is involved in the biosynthesis of L-threo-3-hydroxy-glutamate, prior to incorporation into kutznerides (antifungal nonribosomal hexadepsipeptides)
725193
Kutzneria sp. 744
S-(threo-3-hydroxy-L-glutamyl)-[peptidyl-carrier-protein of nonribosomal peptide synthetase KtzH] + succinate + CO2
-
-
-
?
S-(L-glutamyl)-[peptidyl-carrier protein of nonribosomal peptide synthetase KtzH] + 2-oxoglutarate + O2
D-Glu, L-Glu and [peptidyl-carrier-protein]-D-glutamyl thioester are not accepted as substrates. Proposed reaction mechanism: the stand-alone adenylation domain KtzN first activates L-glutamic acid as amino acyl adenylate, which then can be transferred site-specifically to the third peptidyl-carrier-protein domain of KtzH. The specificity of this interaction is mediated by the presence of the truncated A domain (A*) as well as hydroxylase KtzO. Both A* and a hydroxylase KtzO are required for Glu-AMP transfer. Subsequently, the PCP-bound glutamic acid is hydroxylated by KtzO to afford L-threo-3-hydroxyglutamic acid
725193
Kutzneria sp. 744
S-(threo-3-hydroxy-L-glutamyl)-[peptidyl-carrier-protein of nonribosomal peptide synthetase KtzH] + succinate + CO2
-
-
-
?
Subunits
Subunits
Commentary
Organism
?
x * 40100, SDS-PAGE
Kutzneria sp. 744
Temperature Optimum [C]
Temperature Optimum [C]
Temperature Optimum Maximum [C]
Commentary
Organism
28
-
assay at
Kutzneria sp. 744
Turnover Number [1/s]
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
additional information
-
additional information
nonhydrolyzable coenzyme A analogs are developed and used to determine the kinetic parameters for KtzO-catalyzed hydroxylation of glutamic acid bound to the carrier protein. To determine the kinetic parameters of KtzO catalyzed hydroxylation, the problem of the labile thioester bond is circumvented by using synthetic coenzyme A analogs coupled to glutamic acid where the thioester is replaced by a hydrolytically stable amide bond
Kutzneria sp. 744
pH Optimum
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7
-
assay at
Kutzneria sp. 744
Cloned(Commentary) (protein specific)
Commentary
Organism
overproduced in Escherichia coli as His7-tagged fusions
Kutzneria sp. 744
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
additional information
-
additional information
nonhydrolyzable coenzyme A analogs are developed and used to determine the kinetic parameters for KtzO-catalyzed hydroxylation of glutamic acid bound to the carrier protein. To determine the kinetic parameters of KtzO catalyzed hydroxylation, the problem of the labile thioester bond is circumvented by using synthetic coenzyme A analogs coupled to glutamic acid where the thioester is replaced by a hydrolytically stable amide bond
Kutzneria sp. 744
Metals/Ions (protein specific)
Metals/Ions
Commentary
Organism
Structure
Iron
putative non-heme iron oxygenase, exhibit the conserved HXD/E_H iron(II)-binding motif, in the assay (NH4)2FeSO4 is used as source of the ferrous iron cofactor (0.5 mM)
Kutzneria sp. 744
Molecular Weight [Da] (protein specific)
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
40100
-
x * 40100, SDS-PAGE
Kutzneria sp. 744
Natural Substrates/ Products (Substrates) (protein specific)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
S-(L-glutamyl)-[peptidyl-carrier protein of nonribosomal peptide synthetase KtzH] + 2-oxoglutarate + O2
Kutzneria sp. 744
the enzyme is involved in the biosynthesis of L-threo-3-hydroxy-glutamate, prior to incorporation into kutznerides (antifungal nonribosomal hexadepsipeptides)
S-(threo-3-hydroxy-L-glutamyl)-[peptidyl-carrier-protein of nonribosomal peptide synthetase KtzH] + succinate + CO2
-
-
?
Purification (Commentary) (protein specific)
Commentary
Organism
-
Kutzneria sp. 744
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
additional information
able to hydroxylate glutamic acid bound to a noncognate peptidyl-carrier-protein, e.g. the ninth peptidyl-carrier-protein domain of the CDA NRPS assembly line (CDA-T9). Nonhydrolyzable coenzyme A analogs are developed and used to determine the kinetic parameters for KtzO-catalyzed hydroxylation of glutamic acid bound to the carrier protein. To determine the kinetic parameters of KtzO catalyzed hydroxylation, the problem of the labile thioester bond is circumvented by using synthetic coenzyme A analogs coupled to glutamic acid where the thioester is replaced by a hydrolytically stable amide bond
725193
Kutzneria sp. 744
?
-
-
-
-
S-(L-glutamyl)-[peptidyl-carrier protein of nonribosomal peptide synthetase KtzH] + 2-oxoglutarate + O2
the enzyme is involved in the biosynthesis of L-threo-3-hydroxy-glutamate, prior to incorporation into kutznerides (antifungal nonribosomal hexadepsipeptides)
725193
Kutzneria sp. 744
S-(threo-3-hydroxy-L-glutamyl)-[peptidyl-carrier-protein of nonribosomal peptide synthetase KtzH] + succinate + CO2
-
-
-
?
S-(L-glutamyl)-[peptidyl-carrier protein of nonribosomal peptide synthetase KtzH] + 2-oxoglutarate + O2
D-Glu, L-Glu and [peptidyl-carrier-protein]-D-glutamyl thioester are not accepted as substrates. Proposed reaction mechanism: the stand-alone adenylation domain KtzN first activates L-glutamic acid as amino acyl adenylate, which then can be transferred site-specifically to the third peptidyl-carrier-protein domain of KtzH. The specificity of this interaction is mediated by the presence of the truncated A domain (A*) as well as hydroxylase KtzO. Both A* and a hydroxylase KtzO are required for Glu-AMP transfer. Subsequently, the PCP-bound glutamic acid is hydroxylated by KtzO to afford L-threo-3-hydroxyglutamic acid
725193
Kutzneria sp. 744
S-(threo-3-hydroxy-L-glutamyl)-[peptidyl-carrier-protein of nonribosomal peptide synthetase KtzH] + succinate + CO2
-
-
-
?
Subunits (protein specific)
Subunits
Commentary
Organism
?
x * 40100, SDS-PAGE
Kutzneria sp. 744
Temperature Optimum [C] (protein specific)
Temperature Optimum [C]
Temperature Optimum Maximum [C]
Commentary
Organism
28
-
assay at
Kutzneria sp. 744
Turnover Number [1/s] (protein specific)
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
additional information
-
additional information
nonhydrolyzable coenzyme A analogs are developed and used to determine the kinetic parameters for KtzO-catalyzed hydroxylation of glutamic acid bound to the carrier protein. To determine the kinetic parameters of KtzO catalyzed hydroxylation, the problem of the labile thioester bond is circumvented by using synthetic coenzyme A analogs coupled to glutamic acid where the thioester is replaced by a hydrolytically stable amide bond
Kutzneria sp. 744
pH Optimum (protein specific)
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7
-
assay at
Kutzneria sp. 744
General Information
General Information
Commentary
Organism
physiological function
the enzyme is involved in the biosynthesis of L-threo-3-hydroxy-glutamate, prior to incorporation into kutznerides (antifungal nonribosomal hexadepsipeptides)
Kutzneria sp. 744
General Information (protein specific)
General Information
Commentary
Organism
physiological function
the enzyme is involved in the biosynthesis of L-threo-3-hydroxy-glutamate, prior to incorporation into kutznerides (antifungal nonribosomal hexadepsipeptides)
Kutzneria sp. 744
KCat/KM [mM/s]
kcat/KM Value [1/mMs-1]
kcat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
additional information
-
additional information
nonhydrolyzable coenzyme A analogs are developed and used to determine the kinetic parameters for KtzO-catalyzed hydroxylation of glutamic acid bound to the carrier protein. To determine the kinetic parameters of KtzO catalyzed hydroxylation, the problem of the labile thioester bond is circumvented by using synthetic coenzyme A analogs coupled to glutamic acid where the thioester is replaced by a hydrolytically stable amide bond
Kutzneria sp. 744
KCat/KM [mM/s] (protein specific)
KCat/KM Value [1/mMs-1]
KCat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
additional information
-
additional information
nonhydrolyzable coenzyme A analogs are developed and used to determine the kinetic parameters for KtzO-catalyzed hydroxylation of glutamic acid bound to the carrier protein. To determine the kinetic parameters of KtzO catalyzed hydroxylation, the problem of the labile thioester bond is circumvented by using synthetic coenzyme A analogs coupled to glutamic acid where the thioester is replaced by a hydrolytically stable amide bond
Kutzneria sp. 744
Other publictions for EC 1.14.11.B17
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)
725193
Strieker
Stereospecific synthesis of th ...
Kutzneria sp. 744
J. Am. Chem. Soc.
131
13523-13530
2009
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