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Literature summary for 2.5.1.28 extracted from

  • Surmacz, L.; Plochocka, D.; Kania, M.; Danikiewicz, W.; Swiezewska, E.
    cis-Prenyltransferase atCPT6 produces a family of very short-chain polyisoprenoids in planta (2014), Biochim. Biophys. Acta, 1841, 240-250.
    View publication on PubMed

Cloned(Commentary)

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

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 [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

Localization Comment Organism GeneOntology No. Textmining
chloroplast
-
Solanum habrochaites 9507
-

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+ required Solanum habrochaites

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
dimethylallyl diphosphate + isopentenyl diphosphate Solanum habrochaites
-
diphosphate + neryl diphosphate
-
?

Organism

Organism UniProt Comment Textmining
Solanum habrochaites W0J9X7 monoterpene-producing accession LA2409, gene NDPS1 of the CPT1 locus
-

Purification (Commentary)

Purification (Comment) Organism
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli by nickel affinity chromatography Solanum habrochaites

Source Tissue

Source Tissue Comment Organism Textmining
glandular trichome type VI glandular trichome Solanum habrochaites
-

Substrates and Products (Substrate)

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

Synonyms Comment Organism
NDPS1
-
Solanum habrochaites
neryl diphosphate synthase 1
-
Solanum habrochaites

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
30
-
assay at Solanum habrochaites

Turnover Number [1/s]

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

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.6
-
assay at Solanum habrochaites

General Information

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 [mM/s]

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