Cloned (Comment) | Organism |
---|---|
overproduced in Escherichia coli as His7-tagged fusions | Kutzneria sp. 744 |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | 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 | Comment | 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 Maximum [Da] | Comment | Organism |
---|---|---|---|
40100 | - |
x * 40100, SDS-PAGE | Kutzneria sp. 744 |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
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 | UniProt | Comment | Textmining |
---|---|---|---|
Kutzneria sp. 744 | A8CF77 | - |
- |
Purification (Comment) | Organism |
---|---|
- |
Kutzneria sp. 744 |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
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 | 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) | 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 | Kutzneria sp. 744 | S-(threo-3-hydroxy-L-glutamyl)-[peptidyl-carrier-protein of nonribosomal peptide synthetase KtzH] + succinate + CO2 | - |
? |
Subunits | Comment | Organism |
---|---|---|
? | x * 40100, SDS-PAGE | Kutzneria sp. 744 |
Synonyms | Comment | Organism |
---|---|---|
KtzO | - |
Kutzneria sp. 744 |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
28 | - |
assay at | Kutzneria sp. 744 |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | 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 Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7 | - |
assay at | Kutzneria sp. 744 |
General Information | Comment | 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 Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | 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 |