Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary extracted from

  • Rudolph, K.; Parthier, C.; Egerer-Sieber, C.; Geiger, D.; Muller, Y.A.; Kreis, W.; Mueller-Uri, F.
    Expression, crystallization and structure elucidation of gamma-terpinene synthase from Thymus vulgaris (2016), Acta Crystallogr. Sect. F, 72, 16-23 .
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
4.2.3.114 gene TvTPS, DNA and amino acid sequence determination and analysis, recombinant expression of His-tagged enzyme in Escherichia coli strain BL21 CodonPlus Thymus vulgaris

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
4.2.3.114 purified recombinant enzyme, sitting drop method, mixing of 400-600 nl of 10 mg/ml protein in 20 mM Tris-HCl, pH 7.8, 1 mM MgCl2, 4 mM DTT, with reservoir solution containing 0.2 M ammonium tartrate, pH 7.0, 20% w/v PEG 3350, in a 1:1 or 1:2 ratio, equilibration against 0.07 ml reservoir solution, 19°C, X-ray diffraction structure determination and analysis at 1.65-1.90 A resolution, two molecules (monomers A and B) in the asymmetric unit, molecular replacement using the crystal structure of (4S)-limonene synthase from Mentha spicata, PDB ID 2ong, model building Thymus vulgaris

Protein Variants

EC Number Protein Variants Comment Organism
4.2.3.114 D356A site-directed mutagenesis, the activity of the mutant is completely abolished Thymus vulgaris
4.2.3.114 T565A site-directed mutagenesis, the mutation results in a 82% decrease in activity compared to wild-type Thymus vulgaris

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
4.2.3.114 Mg2+ required Thymus vulgaris

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
4.2.3.114 geranyl diphosphate Thymus vulgaris
-
 gamma-terpinene + diphosphate
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
4.2.3.114 Thymus vulgaris A0A0M3Q1Q3 isozyme TvTPS1
-
4.2.3.114 Thymus vulgaris K9Y6Y9 isozyme TvTPS2
-

Purification (Commentary)

EC Number Purification (Comment) Organism
4.2.3.114 recombinant His-tagged enzyme from Escherichia coli strain BL21 CodonPlus by nickel affinity chromatography, ultrafiltration, and gel filtration Thymus vulgaris

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4.2.3.114 geranyl diphosphate
-
 Thymus vulgaris gamma-terpinene + diphosphate
-
 ?
4.2.3.114 additional information product identification and quantification by GC-MS analysis Thymus vulgaris ?
-
 ?

Subunits

EC Number Subunits Comment Organism
4.2.3.114 dimer overall structure of each TvTPS1 monomer comprises two alpha-helical domains: an N-terminal domain comprising an alpha/alpha barrel and a C-terminal orthogonal bundle domain, crystal structure analysis Thymus vulgaris

Synonyms

EC Number Synonyms Comment Organism
4.2.3.114 TPS1
-
 Thymus vulgaris
4.2.3.114 TPS2
-
 Thymus vulgaris
4.2.3.114 TvTPS
-
 Thymus vulgaris
4.2.3.114 TvTPS1
-
 Thymus vulgaris
4.2.3.114 TvTPS2
-
 Thymus vulgaris

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
4.2.3.114 25
-
 assay at Thymus vulgaris

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
4.2.3.114 7
-
 assay at Thymus vulgaris

General Information

EC Number General Information Comment Organism
4.2.3.114 additional information putative active site of TvTPS is deduced from its pronounced structural similarity to enzymes from other species of the Lamiaceae family involved in terpenoid biosynthesis Thymus vulgaris
4.2.3.114 physiological function The biosynthesis of gamma-terpinene, a precursor of the phenolic isomers thymol and carvacrol found in the essential oil from Thymus sp., is attributed to the activitiy of gamma-terpinene synthase (TvTPS) Thymus vulgaris