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Literature summary extracted from

  • Kumeta, Y.; Ito, M.
    Characterization of delta-guaiene synthases from cultured cells of Aquilaria, responsible for the formation of the sesquiterpenes in agarwood (2010), Plant Physiol., 154, 1998-2007.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
4.2.3.87 expressed in Escherichia coli BL21 (DE3) Codon Plus-RIL cells Aquilaria crassna
4.2.3.93 expressed in Escherichia coli BL21 (DE) Codon plus-RIL cells Aquilaria crassna

Protein Variants

EC Number Protein Variants Comment Organism
4.2.3.87 W144R/V241E/P296S/H337Y inactive Aquilaria crassna
4.2.3.87 Y110S/V241E/P296S/H337Y the mutant produces 34.1% alpha-guaiene, 1.3% alpha-humulene and 64.6% delta-guaiene Aquilaria crassna
4.2.3.87 Y110S/W144R/P296S/H337Y inactive Aquilaria crassna
4.2.3.87 Y110S/W144R/V241E/H337Y inactive Aquilaria crassna
4.2.3.87 Y110S/W144R/V241E/P296S inactive Aquilaria crassna
4.2.3.87 Y110S/W144R/V241E/P296S/H337Y the mutant produces 31.1% alpha-guaiene, 1.5% alpha-humulene and 67.4% delta-guaiene Aquilaria crassna
4.2.3.93 Y110S/V241E/P296S/H337Y the mutant shows decreased catalytic efficiency compared to the wild type enzyme Aquilaria crassna
4.2.3.93 Y110S/W144R/V241E/P296S/H337Y the mutant shows decreased catalytic efficiency compared to the wild type enzyme Aquilaria crassna

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
4.2.3.93 0.000685
-
 (2E,6E)-farnesyl diphosphate mutant enzyme Y110S/V241E/P296S/H337Y, in Tris-HCl (25 mM, pH 7.0), at 30°C Aquilaria crassna
4.2.3.93 0.00271
-
 (2E,6E)-farnesyl diphosphate wild type enzyme, in 50 mM Tris-HCl, pH 7.5, at 30°C Aquilaria crassna
4.2.3.93 0.00429
-
 (2E,6E)-farnesyl diphosphate mutant enzyme Y110S/W144R/V241E/P296S/H337Y, in Tris-HCl (25 mM, pH 7.0), at 30°C Aquilaria crassna

Organism

EC Number Organism UniProt Comment Textmining
4.2.3.87 Aquilaria crassna
-
-
-
4.2.3.93 Aquilaria crassna
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
4.2.3.87 Ni-NTA column chromatography, gel filtration Aquilaria crassna
4.2.3.93 Ni-NTA column chromatography Aquilaria crassna

Source Tissue

EC Number Source Tissue Comment Organism Textmining
4.2.3.87 leaf
-
 Aquilaria crassna
-
4.2.3.93 cell culture
-
 Aquilaria crassna
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4.2.3.87 (2E,6E)-farnesyl diphosphate
-
 Aquilaria crassna alpha-guaiene + diphosphate 13% yield ?
4.2.3.93 (2E,6E)-farnesyl diphosphate
-
 Aquilaria crassna delta-guaiene + diphosphate delta-guaiene is the major product (61% yield) ?

Synonyms

EC Number Synonyms Comment Organism
4.2.3.87 TPS
-
 Aquilaria crassna
4.2.3.93 TPS
-
 Aquilaria crassna

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
4.2.3.93 0.000586
-
 (2E,6E)-farnesyl diphosphate mutant enzyme Y110S/W144R/V241E/P296S/H337Y, in Tris-HCl (25 mM, pH 7.0), at 30°C Aquilaria crassna
4.2.3.93 0.00125
-
 (2E,6E)-farnesyl diphosphate mutant enzyme Y110S/V241E/P296S/H337Y, in Tris-HCl (25 mM, pH 7.0), at 30°C Aquilaria crassna
4.2.3.93 0.00499
-
 (2E,6E)-farnesyl diphosphate wild type enzyme, in 50 mM Tris-HCl, pH 7.5, at 30°C Aquilaria crassna

Expression

EC Number Organism Comment Expression
4.2.3.87 Aquilaria crassna the production of alpha-guaiene is induced by treatment with methyl jasmonate up
4.2.3.93 Aquilaria crassna the production of delta-guaiene is induced by treatment with 300 mM methyl jasmonate reaching its maximum after 12 h up

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
4.2.3.93 0.137
-
 (2E,6E)-farnesyl diphosphate mutant enzyme Y110S/W144R/V241E/P296S/H337Y, in Tris-HCl (25 mM, pH 7.0), at 30°C Aquilaria crassna
4.2.3.93 0.182
-
 (2E,6E)-farnesyl diphosphate mutant enzyme Y110S/V241E/P296S/H337Y, in Tris-HCl (25 mM, pH 7.0), at 30°C Aquilaria crassna
4.2.3.93 1.836
-
 (2E,6E)-farnesyl diphosphate wild type enzyme, in 50 mM Tris-HCl, pH 7.5, at 30°C Aquilaria crassna