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Literature summary extracted from

  • Zhou, K.; Gao, Y.; Hoy, J.A.; Mann, F.M.; Honzatko, R.B.; Peters, R.J.
    Insights into diterpene cyclization from structure of bifunctional abietadiene synthase from Abies grandis (2012), J. Biol. Chem., 287, 6840-6850.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
4.2.3.18 expressed in Escherichia coli C41 OverExpress and B834(DE3) cells Abies grandis
4.2.3.132 DELTA84 pseudo-mature form of abietadiene synthase is expressed in Escherichia coli C41 and B834(DE3) cells Abies grandis

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
4.2.3.18 hanging drop vapor diffusion method, using 24% (v/v) PEG 8000, 0.1 M sodium citrate, pH 5.1, 0.1 M dibasic ammonium phosphate Abies grandis
4.2.3.132 hanging drop vapor diffusion method, using 24% (w/v) PEG 8000, 0.1 M sodium citrate, pH 5.1, 0.1 M dibasic ammonium phosphate Abies grandis

Protein Variants

EC Number Protein Variants Comment Organism
4.2.3.18 N451A this mutation reduces class II (protonation-initiated cyclization) activity about 100fold, with essentially no effect on class I (ionization-initiated cyclization) activity Abies grandis
4.2.3.132 D621A the mutation selectively abrogates abietadiene synthase class I activity Abies grandis
4.2.3.132 N451A the mutant shows reduced class I abietadiene synthase activity compared to the wild type enzyme Abies grandis
4.2.3.132 R356A the mutant shows reduced class I abietadiene synthase activity compared to the wild type enzyme Abies grandis

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
4.2.3.132 0.0007
-
(+)-copalyl diphosphate mutant enzyme R356A, pH and temperature not specified in the publication Abies grandis
4.2.3.132 0.0014
-
(+)-copalyl diphosphate mutant enzyme N451A, pH and temperature not specified in the publication Abies grandis
4.2.3.132 0.002
-
(+)-copalyl diphosphate wild type enzyme, pH and temperature not specified in the publication 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
-
abieta-7,13-diene + diphosphate
-
?

Organism

EC Number Organism UniProt Comment Textmining
4.2.3.18 Abies grandis Q38710
-
-
4.2.3.132 Abies grandis Q38710
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
4.2.3.18 Mono Q column chromatography and type II ceramic hydroxyapatite column chromatography Abies grandis
4.2.3.132 type II ceramic hydroxyapatite column chromatography and Mono Q column chromatography Abies grandis

Storage Stability

EC Number Storage Stability Organism
4.2.3.18 -80°C, purified enzyme in 10 mM Bis-Tris, pH 6.8, 10% (v/v) glycerol,150 mM KCl, 10 mM MgCl2, and 5 mM dithiohtreitol, several months, without significant loss of activity Abies grandis
4.2.3.132 -80°C, 10 mM Bis-Tris, pH 6.8, 10% (v/v) glycerol, 150 mM KCl, 10 mM MgCl2, and 5 mM dithiothreitol, several months, without significant loss of activity Abies grandis

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4.2.3.18 (+)-copalyl diphosphate
-
Abies grandis abieta-7,13-diene + diphosphate
-
?
4.2.3.132 (+)-copalyl diphosphate
-
Abies grandis neoabietadiene + diphosphate
-
?

Subunits

EC Number Subunits Comment Organism
4.2.3.132 homotetramer x-ray crystallography Abies grandis

Synonyms

EC Number Synonyms Comment Organism
4.2.3.18 abietadiene synthase
-
Abies grandis
4.2.3.132 abietadiene synthase cf. EC 4.2.3.18 Abies grandis

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
4.2.3.132 1.1
-
(+)-copalyl diphosphate mutant enzyme R356A, pH and temperature not specified in the publication Abies grandis
4.2.3.132 1.9
-
(+)-copalyl diphosphate mutant enzyme N451A, pH and temperature not specified in the publication Abies grandis
4.2.3.132 2.6
-
(+)-copalyl diphosphate wild type enzyme, pH and temperature not specified in the publication Abies grandis