Cloned (Comment) | Organism |
---|---|
gene NDPS1 of the CPT1 locus, DNA and amino acid sequence determination and analysis, sequence comparisons and phylogenetic analysis, functional expression of codon-optimized synthetic version of CPT1 of accession LA2409, a monoterpene-producing accession, recombinant expression of His-tagged wild-type and mutant enzymes in Escherichia coli | Solanum habrochaites |
Protein Variants | Comment | Organism |
---|---|---|
E98D/Y100S/I106L/I107F | site-directed mutagenesis, mutant M4 | Solanum habrochaites |
E98D/Y100S/I107F | site-directed mutagenesis, mutant M3V2 | Solanum habrochaites |
additional information | change of 10 residues that correlate with changes in substrate in NDPS1 from accession LA2409 to those found in zFPS, EC 2.5.1.92, from accession LA1393 (Pro/Ser-45, which corresponds to the second residue in the predicted mature protein, is excluded), and reciprocal changes in zFPS (LA1393). The two resulting recombinant proteins are named NDPS1-M10 and zFPS-M10. The NDPS1-M10 mutant variant is able to utilize dimethylallyl diphosphate and isopentenyl diphosphate to synthesize Z,Z-farnesyl diphosphate at levels that are similar to that of the zFPS enzyme from accession LA1393. The ability of mutant NDPS1-M10 to synthesize Z,Z-farnesyl diphosphate coincides with a greatly reduced ability to synthesize neryl diphosphate | Solanum habrochaites |
Y100S/I106L/I107F | site-directed mutagenesis, mutant M3V1 | Solanum habrochaites |
Y100S/I107F | site-directed mutagenesis, mutant M2 | Solanum habrochaites |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.014 | - |
neryl diphosphate | pH 7.6, 30°C, mutant NDPS1-M10 | Solanum habrochaites | |
0.047 | - |
dimethylallyl diphosphate | pH 7.6, 30°C, mutant NDPS1-M10 | Solanum habrochaites | |
0.054 | - |
dimethylallyl diphosphate | pH 7.6, 30°C, mutant NDPS1-M4 | Solanum habrochaites | |
0.097 | - |
neryl diphosphate | pH 7.6, 30°C, mutant NDPS1-M4 | Solanum habrochaites | |
0.111 | - |
isopentenyl diphosphate | pH 7.6, 30°C, mutant NDPS1-M4 | Solanum habrochaites | |
0.112 | - |
dimethylallyl diphosphate | pH 7.6, 30°C, wild-type enzyme | Solanum habrochaites | |
0.14 | - |
isopentenyl diphosphate | pH 7.6, 30°C, wild-type enzyme | Solanum habrochaites | |
0.157 | - |
isopentenyl diphosphate | pH 7.6, 30°C, mutant NDPS1-M10 | Solanum habrochaites |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
chloroplast | - |
Solanum habrochaites | 9507 | - |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | required | Solanum habrochaites |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
dimethylallyl diphosphate + isopentenyl diphosphate | Solanum habrochaites | - |
diphosphate + neryl diphosphate | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Solanum habrochaites | W0J9X7 | monoterpene-producing accession LA2409, gene NDPS1 of the CPT1 locus | - |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli by nickel affinity chromatography | Solanum habrochaites |
Source Tissue | Comment | Organism | Textmining |
---|---|---|---|
glandular trichome | type VI glandular trichome | Solanum habrochaites | - |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
dimethylallyl diphosphate + isopentenyl diphosphate | - |
Solanum habrochaites | diphosphate + neryl diphosphate | - |
? | |
additional information | the relative positioning of aromatic amino acid residues at positions 100 and 107 determines the ability of these enzymes to synthesize neryl diphosphate or (2Z,6Z)-farnesyl diphosphate | Solanum habrochaites | ? | - |
? | |
neryl diphosphate + isopentenyl diphosphate | mutant NDPS1-M10 | Solanum habrochaites | diphosphate + (2Z,6Z)-farnesyl diphosphate | - |
? |
Synonyms | Comment | Organism |
---|---|---|
NDPS1 | - |
Solanum habrochaites |
neryl diphosphate synthase 1 | - |
Solanum habrochaites |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
assay at | Solanum habrochaites |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0064 | - |
dimethylallyl diphosphate | pH 7.6, 30°C, mutant NDPS1-M10 | Solanum habrochaites | |
0.011 | - |
isopentenyl diphosphate | pH 7.6, 30°C, mutant NDPS1-M10 | Solanum habrochaites | |
0.026 | - |
dimethylallyl diphosphate | pH 7.6, 30°C, wild-type enzyme | Solanum habrochaites | |
0.047 | - |
isopentenyl diphosphate | pH 7.6, 30°C, wild-type enzyme | Solanum habrochaites | |
0.088 | - |
dimethylallyl diphosphate | pH 7.6, 30°C, mutant NDPS1-M4 | Solanum habrochaites | |
0.16 | - |
isopentenyl diphosphate | pH 7.6, 30°C, mutant NDPS1-M4 | Solanum habrochaites | |
0.18 | - |
neryl diphosphate | pH 7.6, 30°C, mutant NDPS1-M4 | Solanum habrochaites | |
3.7 | - |
neryl diphosphate | pH 7.6, 30°C, mutant NDPS1-M10 | Solanum habrochaites |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.6 | - |
assay at | Solanum habrochaites |
General Information | Comment | Organism |
---|---|---|
evolution | biochemical evolution of terpene biosynthesis in the glandular trichomes of Solanum species, comparative sequence analysis and evolutionary relationship of NDPS1 and (2Z,6Z)-farnesyl diphosphate synthase (EC 2.5.1.92, zFPS), cDNAs from the CPT1 locus, phylogenetic analysis, overview | Solanum habrochaites |
additional information | homology modeling and three-dimensional structure | Solanum habrochaites |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.15 | - |
dimethylallyl diphosphate | pH 7.6, 30°C, mutant NDPS1-M10 | Solanum habrochaites | |
0.23 | - |
dimethylallyl diphosphate | pH 7.6, 30°C, wild-type enzyme | Solanum habrochaites | |
0.28 | - |
neryl diphosphate | pH 7.6, 30°C, mutant NDPS1-M10 | Solanum habrochaites | |
0.33 | - |
isopentenyl diphosphate | pH 7.6, 30°C, wild-type enzyme | Solanum habrochaites | |
0.74 | - |
isopentenyl diphosphate | pH 7.6, 30°C, mutant NDPS1-M10 | Solanum habrochaites | |
1.6 | - |
dimethylallyl diphosphate | pH 7.6, 30°C, mutant NDPS1-M4 | Solanum habrochaites | |
1.8 | - |
isopentenyl diphosphate | pH 7.6, 30°C, mutant NDPS1-M4 | Solanum habrochaites | |
1.8 | - |
neryl diphosphate | pH 7.6, 30°C, mutant NDPS1-M4 | Solanum habrochaites |