Cloned (Comment) | Organism |
---|---|
expression of His4-tagged CURS1 in Escherichia coli strain BL21(DE3) | Curcuma longa |
Crystallization (Comment) | Organism |
---|---|
purified recombinant His4-tagged CURS1, sitting drop vapor diffusion method, mixing of 10 mg/ml protein in 10 mM Tris-HCl, pH 8.0, 10% glycerol, and 1 mM tris(2-carboxyethyl)phosphine with reservoir solution containing 0.3 M sodium malonate, pH 7.0, and 25% PEG3350, X-ray diffraction structure determination and analysis at 2.32 A resolution, modeling | Curcuma longa |
Protein Variants | Comment | Organism |
---|---|---|
G211F | site-directed mutagenesis, the mutant shows highly reduced 2-oxoacid condensation activity compared to the wild-type enzyme | Curcuma longa |
H303A | site-directed mutagenesis, active site residue mutant, the mutant shows reduced 2-oxoacid condensation activity compared to the wild-type enzyme | Curcuma longa |
H303Q | site-directed mutagenesis, active site residue mutant, the mutant shows reduced 2-oxoacid condensation activity compared to the wild-type enzyme | Curcuma longa |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0012 | - |
3-oxo-5-phenyl-4-pentenoic acid | pH 7.5, 37°C, recombinant mutant H303Q | Curcuma longa | |
0.0017 | - |
cinnamoyl-diketide-N-acetylcysteamine | pH 7.5, 37°C, recombinant wild-type enzyme | Curcuma longa | |
0.003 | - |
cinnamoyl-diketide-N-acetylcysteamine | pH 7.5, 37°C, recombinant mutant H303Q | Curcuma longa | |
0.0059 | - |
3-oxo-5-phenyl-4-pentenoic acid | pH 7.5, 37°C, recombinant wild-type enzyme | Curcuma longa | |
0.113 | - |
cinnamoyl-diketide-N-acetylcysteamine | pH 7.5, 37°C, recombinant mutant G211F | Curcuma longa | |
0.138 | - |
3-oxo-5-phenyl-4-pentenoic acid | pH 7.5, 37°C, recombinant mutant G211F | Curcuma longa |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
feruloyl-CoA + cinnamoyl-diketide-N-acetylcysteamine + H2O | Curcuma longa | - |
CoA + cinnamoylferuloylmethane + N-acetylcysteamine + CO2 | - |
? | |
feruloyl-CoA + feruloylacetyl-CoA + H2O | Curcuma longa | - |
2 CoA + curcumin + CO2 | - |
? | |
additional information | Curcuma longa | CURS1 catalyzes the hydrolysis of diketide-CoA to yield a 2-oxoacid (ii) and decarboxylative condensation of the 2-oxoacid with feruloyl-CoA to yield curcumin | ? | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Curcuma longa | C0SVZ6 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant His4-tagged CURS1 from Escherichia coli strain BL21(DE3) by nickel affinity and anion exchange chromatography, followed by ultrafiltration | Curcuma longa |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
feruloyl-CoA + 3-oxo-5-phenyl-4-pentenoic acid + H2O | - |
Curcuma longa | CoA + cinnamoylferuloylmethane + CO2 | - |
? | |
feruloyl-CoA + cinnamoyl-diketide-N-acetylcysteamine + H2O | - |
Curcuma longa | CoA + cinnamoylferuloylmethane + N-acetylcysteamine + CO2 | - |
? | |
feruloyl-CoA + cinnamoyl-diketide-N-acetylcysteamine + H2O | via 3-oxo-5-phenyl-4-pentenoic acid | Curcuma longa | CoA + cinnamoylferuloylmethane + N-acetylcysteamine + CO2 | - |
? | |
feruloyl-CoA + feruloylacetyl-CoA + H2O | - |
Curcuma longa | 2 CoA + curcumin + CO2 | - |
? | |
feruloyl-CoA + feruloylacetyl-CoA + H2O | two-step reaction of hydrolysis and decarboxylative condensation | Curcuma longa | 2 CoA + curcumin + CO2 | - |
? | |
additional information | CURS1 catalyzes the hydrolysis of diketide-CoA to yield a 2-oxoacid (ii) and decarboxylative condensation of the 2-oxoacid with feruloyl-CoA to yield curcumin | Curcuma longa | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
curcumin synthase 1 | - |
Curcuma longa |
CURS1 | - |
Curcuma longa |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Curcuma longa |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.00007 | - |
cinnamoyl-diketide-N-acetylcysteamine | pH 7.5, 37°C, recombinant mutant H303Q | Curcuma longa | |
0.0015 | - |
cinnamoyl-diketide-N-acetylcysteamine | pH 7.5, 37°C, recombinant mutant G211F | Curcuma longa | |
0.0017 | - |
3-oxo-5-phenyl-4-pentenoic acid | pH 7.5, 37°C, recombinant mutant H303Q | Curcuma longa | |
0.0043 | - |
cinnamoyl-diketide-N-acetylcysteamine | pH 7.5, 37°C, recombinant wild-type enzyme | Curcuma longa | |
0.0083 | - |
3-oxo-5-phenyl-4-pentenoic acid | pH 7.5, 37°C, recombinant mutant G211F | Curcuma longa | |
0.125 | - |
3-oxo-5-phenyl-4-pentenoic acid | pH 7.5, 37°C, recombinant wild-type enzyme | Curcuma longa |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | 8 | assay at | Curcuma longa |
General Information | Comment | Organism |
---|---|---|
evolution | the enzyme belongs to the Type III polyketide synthase family | Curcuma longa |
metabolism | the enzyme catalyzes a step in the curcuminoid biosynthesis, pathway overview | Curcuma longa |
additional information | active site residue His303 is important for 2-oxoacid condensation, involvement of the hydrophobic cavity around Phe265 in 2-oxoacid condensation, wild-type and mutant ligand binding structures and structure-function relationship, modeling, overview | Curcuma longa |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.013 | - |
cinnamoyl-diketide-N-acetylcysteamine | pH 7.5, 37°C, recombinant mutant G211F | Curcuma longa | |
0.024 | - |
cinnamoyl-diketide-N-acetylcysteamine | pH 7.5, 37°C, recombinant mutant H303Q | Curcuma longa | |
0.112 | - |
3-oxo-5-phenyl-4-pentenoic acid | pH 7.5, 37°C, recombinant mutant G211F | Curcuma longa | |
1.605 | - |
3-oxo-5-phenyl-4-pentenoic acid | pH 7.5, 37°C, recombinant mutant H303Q | Curcuma longa | |
2.915 | - |
cinnamoyl-diketide-N-acetylcysteamine | pH 7.5, 37°C, recombinant wild-type enzyme | Curcuma longa | |
20.54 | - |
3-oxo-5-phenyl-4-pentenoic acid | pH 7.5, 37°C, recombinant wild-type enzyme | Curcuma longa |