Literature summary extracted from

  • Koeduka, T.; Baiga, T.J.; Noel, J.P.; Pichersky, E.
    Biosynthesis of t-anethole in anise: characterization of t-anol/isoeugenol synthase and an O-methyltransferase specific for a C7-C8 propenyl side chain (2009), Plant Physiol., 149, 384-394.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.1.1.146 expression in Escherichia coli Pimpinella anisum
1.1.1.319 DNA and amino acid sequence determination and analysis, phylogenetic analysis and tree, expression in Escherichia coli Pimpinella anisum
2.1.1.279 expression in Escherichia coli Pimpinella anisum

Inhibitors

EC Number Inhibitors Comment Organism Structure
1.1.1.319 Ca2+ weak inhibition at 2.5 mM Pimpinella anisum
1.1.1.319 Cu2+ strong inhibition at 2.5 mM Pimpinella anisum
1.1.1.319 Fe2+ strong inhibition at 2.5 mM Pimpinella anisum
1.1.1.319 K+ weak inhibition at 2.5 mM Pimpinella anisum
1.1.1.319 Mn2+ weak inhibition at 2.5 mM Pimpinella anisum
1.1.1.319 additional information no inhibitory effect on the activity with 2.5 mM Na+ or Mg2+ Pimpinella anisum
1.1.1.319 Zn2+ strong inhibition at 2.5 mM Pimpinella anisum
2.1.1.146 Cu2+ 2.5 mM, 97% inhibition Pimpinella anisum
2.1.1.146 Fe2+ 2.5 mM, 47% inhibition Pimpinella anisum
2.1.1.146 Mn2+ 2.5 mM, 35% inhibition Pimpinella anisum
2.1.1.146 additional information not inhibitory: 2.5 mM K+, Na+, Ca2+, and Mg2+ Pimpinella anisum
2.1.1.146 Zn2+ 2.5 mM, 97% inhibition Pimpinella anisum
2.1.1.279 Cu2+ 2.5 mM, 97% inhibition Pimpinella anisum
2.1.1.279 Fe2+ 2.5 mM, 47% inhibition Pimpinella anisum
2.1.1.279 Mn2+ 2.5 mM, 35% inhibition Pimpinella anisum
2.1.1.279 additional information no inhibitory effect on the activity is observed by incubation with 2.5 mM K+, Na+, Ca2+, and Mg2+ Pimpinella anisum
2.1.1.279 Zn2+ 2.5 mM, 97% inhibition Pimpinella anisum

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.1.1.319 0.23
-
coniferyl acetate pH 7.5, 25°C Pimpinella anisum
2.1.1.146 0.0193
-
isoeugenol pH 7.5, 25°C Pimpinella anisum
2.1.1.279 0.0193
-
isoeugenol pH 7.5, 25°C Pimpinella anisum
1.1.1.319 0.134
-
4-coumaryl acetate pH 7.5, 25°C Pimpinella anisum
2.1.1.146 0.0545
-
S-adenosyl-L-methionine pH 7.5, 25°C Pimpinella anisum
2.1.1.279 0.0545
-
S-adenosyl-L-methionine pH 7.5, 25°C Pimpinella anisum

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.1.1.319 4-coumaryl acetate + NADPH + H+ Pimpinella anisum
-
t-anol + acetate + NADP+
-
?
1.1.1.319 coniferyl acetate + NADPH + H+ Pimpinella anisum
-
isoeugenol + acetate + NADP+
-
?

Organism

EC Number Organism UniProt Comment Textmining
2.1.1.146 Pimpinella anisum
-
-
-
1.1.1.319 Pimpinella anisum B8RCD2
-
-
2.1.1.279 Pimpinella anisum B8RCD3
-
-

Source Tissue

EC Number Source Tissue Comment Organism Textmining
2.1.1.146 additional information expression throughout the plant with highest level in developing fruit Pimpinella anisum
-
2.1.1.146 fruit expression throughout the plant with highest level in developing fruit Pimpinella anisum
-
1.1.1.319 additional information the gene encoding AIS1 is expressed throughout the plant, its transcript level is highest in developing fruits Pimpinella anisum
-
1.1.1.319 fruit developing Pimpinella anisum
-
2.1.1.279 root
-
Pimpinella anisum
-
2.1.1.279 leaf young, very low activity in mature leaves Pimpinella anisum
-
2.1.1.279 fruit the gene encoding AIMT1 is expressed throughout the plant and the transcript level is highest in developing fruits Pimpinella anisum
-
2.1.1.279 flower
-
Pimpinella anisum
-
2.1.1.279 bud
-
Pimpinella anisum
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.1.1.146 additional information no substrate: chavicol, eugenol, guaiacol, caffeic acid, 2-propylphenol Pimpinella anisum ?
-
?
2.1.1.279 additional information the enzyme does not have activity with eugenol or chavicol Pimpinella anisum ?
-
?
2.1.1.279 S-adenosyl-L-methionine + isoeugenol the enzyme prefers isoeugenol to t-anol by a factor of 2. The enzyme is highly specific for substrates in which the double bond in the propenyl side chain is located between C7 and C8 Pimpinella anisum S-adenosyl-L-homocysteine + isomethyleugenol
-
?
2.1.1.146 S-adenosyl-L-methionine + isoeugenol methylation of the p-OH group Pimpinella anisum S-adenosyl-L-homocysteine + methylisoeugenol
-
?
2.1.1.279 S-adenosyl-L-methionine + shows a 10fold preference for isoeugenol over eugenol The enzyme is highly specific for substrates in which the double bond in the propenyl side chain is located between C7 and C8 Pimpinella anisum S-adenosyl-L-homocysteine + ?
-
?
2.1.1.146 S-adenosyl-L-methionine + 5-methoxyeugenol about 18% of the activity with isoeugenol Pimpinella anisum S-adenosyl-L-homocysteine + ?
-
?
2.1.1.146 S-adenosyl-L-methionine + dihydrochavicol about 23% of the activity with isoeugenol Pimpinella anisum S-adenosyl-L-homocysteine + ?
-
?
2.1.1.146 S-adenosyl-L-methionine + dihydroeugenol about 50% of the activity with isoeugenol Pimpinella anisum S-adenosyl-L-homocysteine + methyldihydroeugenol
-
?
2.1.1.146 S-adenosyl-L-methionine + t-anol methylation of the p-OH group. 45% of the rate with isoeugenol Pimpinella anisum S-adenosyl-L-homocysteine + t-anethole
-
?
1.1.1.319 4-coumaryl acetate + NADPH + H+
-
Pimpinella anisum t-anol + acetate + NADP+
-
?
1.1.1.319 coniferyl acetate + NADPH + H+
-
Pimpinella anisum isoeugenol + acetate + NADP+
-
?
2.1.1.279 S-adenosyl-L-methionine + chavicol the enzyme shows a 10fold preference for t-anol over chavicol. The enzyme is highly specific for substrates in which the double bond in the propenyl side chain is located between C7 and C8 Pimpinella anisum S-adenosyl-L-homocysteine + ?
-
?
2.1.1.279 S-adenosyl-L-methionine + trans-anol trans-anol i.e. 4-[(1E)-prop-1-en-1-yl]phenol. The enzyme prefers isoeugenol to t-anol by a factor of 2. It shows a 10fold preference for t-anol over chavicol. The enzyme is highly specific for substrates in which the double bond in the propenyl side chain is located between C7 and C8 Pimpinella anisum S-adenosyl-L-homocysteine + trans-anethole trans-anethole i.e. 1-methoxy-4-[(1E)-prop-1-en-1-yl]benzene ?

Synonyms

EC Number Synonyms Comment Organism
2.1.1.146 AIMT1
-
Pimpinella anisum
2.1.1.279 AIMT1
-
Pimpinella anisum
1.1.1.319 IGS1
-
Pimpinella anisum
1.1.1.319 t-anol/isoeugenol synthase 1
-
Pimpinella anisum
1.1.1.319 AIS1
-
Pimpinella anisum
1.1.1.319 PaAIS1
-
Pimpinella anisum
2.1.1.279 S-adenosyl-L-methionine:t-anol/isoeugenol O-methyltransferase
-
Pimpinella anisum
2.1.1.279 t-anol O-methyltransferase
-
Pimpinella anisum

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.1.1.319 25
-
assay at Pimpinella anisum

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
2.1.1.146 37
-
stable for at least 30 min Pimpinella anisum
1.1.1.319 37
-
purified recombinant enzyme, 30 min, 85% activity remaining Pimpinella anisum
2.1.1.146 50
-
30 min, 88% residual activity Pimpinella anisum
1.1.1.319 50
-
purified recombinant enzyme, 30 min, nelow 45 activity remaining Pimpinella anisum
2.1.1.279 50
-
30 min, 12% loss of activity Pimpinella anisum
2.1.1.146 65
-
30 min, less than 5% residual activity Pimpinella anisum
2.1.1.279 65
-
30 min, less than 5% of activity remaining Pimpinella anisum
2.1.1.146 20
-
stable for at least 30 min Pimpinella anisum
1.1.1.319 20
-
purified recombinant enzyme, 30 min, stable Pimpinella anisum
2.1.1.279 20 37 30 min, stable Pimpinella anisum

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.1.1.146 0.015
-
isoeugenol pH 7.5, 25°C Pimpinella anisum
1.1.1.319 0.07
-
4-coumaryl acetate pH 7.5, 25°C Pimpinella anisum
1.1.1.319 1.02
-
coniferyl acetate pH 7.5, 25°C Pimpinella anisum
2.1.1.279 0.015
-
isoeugenol pH 7.5, 25°C Pimpinella anisum

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.1.1.146 7.5 8
-
Pimpinella anisum
1.1.1.319 7.5
-
assay at Pimpinella anisum
2.1.1.279 7.5 8 Tris buffer Pimpinella anisum

pH Range

EC Number pH Minimum pH Maximum Comment Organism
2.1.1.146 6 9
-
Pimpinella anisum
2.1.1.279 6 9 active in the range Pimpinella anisum

Cofactor

EC Number Cofactor Comment Organism Structure
1.1.1.319 NADPH dependent on Pimpinella anisum

General Information

EC Number General Information Comment Organism
1.1.1.319 physiological function t-anol/isoeugenol synthase is an NADPH-dependent enzyme that can biosynthesize t-anol and isoeugenol from coumaryl acetate and coniferyl acetate, respectively Pimpinella anisum

KCat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
2.1.1.146 0.78
-
isoeugenol pH 7.5, 25°C Pimpinella anisum
2.1.1.279 0.78
-
isoeugenol pH 7.5, temperature not specified in the publication Pimpinella anisum
1.1.1.319 0.52
-
4-coumaryl acetate pH 7.5, 25°C Pimpinella anisum
1.1.1.319 4.44
-
coniferyl acetate pH 7.5, 25°C Pimpinella anisum