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Literature summary for 4.2.3.24 extracted from

  • Li, Z.; Gao, R.; Hao, Q.; Zhao, H.; Cheng, L.; He, F.; Liu, L.; Liu, X.; Chou, W.K.; Zhu, H.; Cane, D.E.
    The T296V mutant of amorpha-4,11-diene synthase is defective in allylic diphosphate isomerization but retains the ability to cyclize the intermediate (3R)-nerolidyl diphosphate to amorpha-4,11-diene (2016), Biochemistry, 55, 6599-6604 .
    View publication on PubMedView publication on EuropePMC
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Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli Rosetta(DE3) cells Artemisia annua

Protein Variants

Protein Variants Comment Organism
T296A the mutant predominantly retains amorphadiene synthase activity by converting (2E,6E)-farnesyl diphosphate to 78% amorpha-4,11-diene and 5% (E)-beta-farnesene Artemisia annua
T296I the mutant converts (2E,6E)-farnesyl diphosphate to 100% (E)-beta-farnesene Artemisia annua
T296L the mutant converts (2E,6E)-farnesyl diphosphate to 35.24% amorpha-4,11-diene and 54.05% (E)-beta-farnesene Artemisia annua
T296S the mutant converts (2E,6E)-farnesyl diphosphate to 90.32% amorpha-4,11-diene and 1.57% (E)-beta-farnesene Artemisia annua
T296V the mutant is defective in allylic diphosphate isomerization but retains the ability to cyclize the intermediate (3R,6E)-nerolidyl diphosphate to amorpha-4,11-diene. The mutant catalyzes the abortive conversion of (2E,6E)-farnesyl diphosphate mainly into the acyclic product (E)-beta-farnesene (88%) Artemisia annua

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0108
-
 (2E,6E)-farnesyl diphosphate mutant enzyme T296A, at pH 7.2 and 30°C Artemisia annua
0.0153
-
 (2E,6E)-farnesyl diphosphate wild type enzyme, at pH 7.2 and 30°C Artemisia annua
0.0198
-
 (2E,6E)-farnesyl diphosphate mutant enzyme T296I, at pH 7.2 and 30°C Artemisia annua
0.0282
-
 (2E,6E)-farnesyl diphosphate mutant enzyme T296V, at pH 7.2 and 30°C Artemisia annua
0.033
-
 (2E,6E)-farnesyl diphosphate mutant enzyme T296S, at pH 7.2 and 30°C Artemisia annua

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+ required Artemisia annua

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
(2E,6E)-farnesyl diphosphate Artemisia annua
-
 amorpha-4,11-diene + diphosphate
-
 ?

Organism

Organism UniProt Comment Textmining
Artemisia annua Q9AR04
-
-

Purification (Commentary)

Purification (Comment) Organism
Ni-NTA column chromatography Artemisia annua

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
(2E,6E)-farnesyl diphosphate
-
 Artemisia annua amorpha-4,11-diene + diphosphate
-
 ?

Synonyms

Synonyms Comment Organism
ADS
-
 Artemisia annua
amorphadiene synthase
-
 Artemisia annua

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.0013
-
 (2E,6E)-farnesyl diphosphate mutant enzyme T296I, at pH 7.2 and 30°C Artemisia annua
0.0015
-
 (2E,6E)-farnesyl diphosphate mutant enzyme T296V, at pH 7.2 and 30°C Artemisia annua
0.0059
-
 (2E,6E)-farnesyl diphosphate mutant enzyme T296A, at pH 7.2 and 30°C Artemisia annua
0.0075
-
 (2E,6E)-farnesyl diphosphate mutant enzyme T296S, at pH 7.2 and 30°C Artemisia annua
0.0163
-
 (2E,6E)-farnesyl diphosphate wild type enzyme, at pH 7.2 and 30°C Artemisia annua

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.05
-
 (2E,6E)-farnesyl diphosphate mutant enzyme T296V, at pH 7.2 and 30°C Artemisia annua
0.07
-
 (2E,6E)-farnesyl diphosphate mutant enzyme T296I, at pH 7.2 and 30°C Artemisia annua
0.23
-
 (2E,6E)-farnesyl diphosphate mutant enzyme T296S, at pH 7.2 and 30°C Artemisia annua
0.55
-
 (2E,6E)-farnesyl diphosphate mutant enzyme T296A, at pH 7.2 and 30°C Artemisia annua
1.06
-
 (2E,6E)-farnesyl diphosphate wild type enzyme, at pH 7.2 and 30°C Artemisia annua