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Literature summary for 4.2.3.132 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
Search for more literature for 4.2.3.132

Cloned(Commentary)

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

Crystallization (Commentary)

Crystallization (Comment) Organism
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

Protein Variants Comment Organism
D621A the mutation selectively abrogates abietadiene synthase class I activity Abies grandis
N451A the mutant shows reduced class I abietadiene synthase activity compared to the wild type enzyme Abies grandis
R356A the mutant shows reduced class I abietadiene synthase activity compared to the wild type enzyme Abies grandis

KM Value [mM]

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

Organism

Organism UniProt Comment Textmining
Abies grandis Q38710
-
-

Purification (Commentary)

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

Storage Stability

Storage Stability Organism
-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)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
(+)-copalyl diphosphate
-
 Abies grandis neoabietadiene + diphosphate
-
 ?

Subunits

Subunits Comment Organism
homotetramer x-ray crystallography Abies grandis

Synonyms

Synonyms Comment Organism
abietadiene synthase cf. EC 4.2.3.18 Abies grandis

Turnover Number [1/s]

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