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Literature summary extracted from

  • Zhou, K.; Peters, R.J.
    Investigating the conservation pattern of a putative second terpene synthase divalent metal binding motif in plants (2009), Phytochemistry, 70, 366-369.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
4.2.3.18 Site-directed mutagenesis is carried out via PCR amplification with overlapping mutagenic primers, and the mutant genes verify by complete sequencing. The resulting wild type and mutant genes are then transferred via directional recombination to the T7-promoter and N-terminal 6his fusion expression vector pDEST17. Use of the pDEST17 vector results in a 25 amino acid residue linker between the 6 His-tag and the cloned protein. Abies grandis
5.5.1.12 Site-directed mutagenesis is carried out via PCR amplification with overlapping mutagenic primers, and the mutant genes verify by complete sequencing. The resulting wild type and mutant genes are then transferred via directional recombination to the T7-promoter and N-terminal 6xhis fusion expression vector pDEST17. Use of the pDEST17 vector results in a 25 amino acid residue linker between the 6 His-tag and the cloned protein. Abies grandis

Protein Variants

EC Number Protein Variants Comment Organism
4.2.3.18 D621A eliminates class I activity Abies grandis
4.2.3.18 T769A is active Abies grandis
4.2.3.18 T769G is active Abies grandis
4.2.3.18 T769S is active Abies grandis
5.5.1.12 D621A eliminates class I activity Abies grandis
5.5.1.12 T769A enzyme activity Abies grandis
5.5.1.12 T769G enzyme activity Abies grandis
5.5.1.12 T769S enzyme activity Abies grandis

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
5.5.1.12 additional information
-
 geranylgeranyl diphosphate mutant T769A, value not determined Abies grandis
5.5.1.12 0.0012
-
 geranylgeranyl diphosphate wild-type Abies grandis
5.5.1.12 0.0015
-
 geranylgeranyl diphosphate mutant T769G Abies grandis
5.5.1.12 0.0015
-
 geranylgeranyl diphosphate mutant T769S Abies grandis

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
4.2.3.18 K+ 100 mM included in assay medium Abies grandis
4.2.3.18 Mg2+ terpene synthases (TPS) require divalent metal ion co-factors, typically magnesium Abies grandis
5.5.1.12 K+ 100 mM included in assay medium Abies grandis
5.5.1.12 Mg2+ terpene synthases (TPS) require divalent metal ion co-factors, typically magnesium Abies grandis

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
4.2.3.18 copalyl diphosphate Abies grandis class I terpene synthase reaction abietadienes + diphosphate
-
 ir
5.5.1.12 geranylgeranyl diphosphate Abies grandis class II terpene synthase reaction copalyl diphosphate
-
 r

Organism

EC Number Organism UniProt Comment Textmining
4.2.3.18 Abies grandis
-
-
-
5.5.1.12 Abies grandis
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
4.2.3.18 purification using Ni-NTA His-bind resin in batch mode Abies grandis
5.5.1.12 AgAS is purified using Ni-NTA His-bind resin in batch mode Abies grandis

Source Tissue

EC Number Source Tissue Comment Organism Textmining
4.2.3.18 cell culture
-
 Abies grandis
-
5.5.1.12 cell culture
-
 Abies grandis
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4.2.3.18 copalyl diphosphate class I terpene synthase reaction Abies grandis abietadienes + diphosphate
-
 ir
5.5.1.12 geranylgeranyl diphosphate class II terpene synthase reaction Abies grandis copalyl diphosphate
-
 r

Synonyms

EC Number Synonyms Comment Organism
4.2.3.18 abietadiene synthase
-
 Abies grandis
4.2.3.18 AGAS
-
 Abies grandis
5.5.1.12 abietadiene synthase class II synthase Abies grandis
5.5.1.12 AGAS
-
 Abies grandis

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
4.2.3.18 30
-
 assay at Abies grandis
5.5.1.12 30
-
 assay at Abies grandis

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
5.5.1.12 additional information
-
 geranylgeranyl diphosphate mutant T769A, value below 10000 Abies grandis
5.5.1.12 0.003
-
 geranylgeranyl diphosphate mutant T769G Abies grandis
5.5.1.12 0.03
-
 geranylgeranyl diphosphate mutant T769S Abies grandis
5.5.1.12 0.4
-
 geranylgeranyl diphosphate wild-type Abies grandis

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
4.2.3.18 7.2
-
 assay at Abies grandis
5.5.1.12 7.2
-
 assay at Abies grandis