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Literature summary for 4.2.3.24 extracted from

  • Fang, X.; Li, J.X.; Huang, J.Q.; Xiao, Y.L.; Zhang, P.; Chen, X.Y.
    Systematic identification of functional residues of Artemisia annua amorpha-4,11-diene synthase (2017), Biochem. J., 474, 2191-2202 .
    View publication on PubMed
Search for more literature for 4.2.3.24

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21(DE3) cells Artemisia annua

Protein Variants

Protein Variants Comment Organism
L374Y the mutant enzyme is inactive toward 1,10 closure Artemisia annua
L374Y/G439S the mutant produces 51.9% bisabolyl-type compounds, including alpha-bisabolol (4.7%), beta-bisabolol (7.4%), and cis-gamma-bisabolene (38.6%) Artemisia annua
L374Y/L404V/L405I/G439S the mutations lead to an enzyme generating 80% monocyclic bisabolyltype sesquiterpenes Artemisia annua
T296V the mutation impairs the ring closure activity almost completely. The mutant produces 93.7% linear (E)-beta-farnesene and 6.3% bicyclical amorpha-4,11-diene compared to the wild type enzyme (90% amorpha-4,11-diene) Artemisia annua
T399L/T447G the mutant shows compromised activity in regioselective deprotonation to yield 34.7 and 37.7% normal and aberrant deprotonation products, respectively Artemisia annua
T399S the mutant with increased catalytic efficiency produces 87.4% amorpha-4,11-diene compared to the wild type enzyme (90%) Artemisia annua
T399S/T447S the mutant with reduced catalytic efficiency produces 72.1% amorpha-4,11-diene compared to the wild type enzyme (90%) Artemisia annua
T447S the mutant with increased catalytic efficiency produces 76.9% amorpha-4,11-diene compared to the wild type enzyme (90%) Artemisia annua

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.00191
-
 (2E,6E)-farnesyl diphosphate mutant enzyme T447S, at pH 7.0 and 30°C Artemisia annua
0.00203
-
 (2E,6E)-farnesyl diphosphate wild type enzyme, at pH 7.0 and 30°C Artemisia annua
0.00256
-
 (2E,6E)-farnesyl diphosphate mutant enzyme T399S, at pH 7.0 and 30°C Artemisia annua
0.00366
-
 (2E,6E)-farnesyl diphosphate mutant enzyme T296V, at pH 7.0 and 30°C Artemisia annua
0.00792
-
 (2E,6E)-farnesyl diphosphate mutant enzyme T399S/T447S, at pH 7.0 and 30°C 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
-
-
-

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

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.0162
-
 (2E,6E)-farnesyl diphosphate mutant enzyme T296V, at pH 7.0 and 30°C Artemisia annua
0.186
-
 (2E,6E)-farnesyl diphosphate wild type enzyme, at pH 7.0 and 30°C Artemisia annua
0.321
-
 (2E,6E)-farnesyl diphosphate mutant enzyme T399S, at pH 7.0 and 30°C Artemisia annua
0.334
-
 (2E,6E)-farnesyl diphosphate mutant enzyme T447S, at pH 7.0 and 30°C Artemisia annua
0.518
-
 (2E,6E)-farnesyl diphosphate mutant enzyme T399S/T447S, at pH 7.0 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
4.4
-
 (2E,6E)-farnesyl diphosphate mutant enzyme T296V, at pH 7.0 and 30°C Artemisia annua
65
-
 (2E,6E)-farnesyl diphosphate mutant enzyme T399S/T447S, at pH 7.0 and 30°C Artemisia annua
92
-
 (2E,6E)-farnesyl diphosphate wild type enzyme, at pH 7.0 and 30°C Artemisia annua
125
-
 (2E,6E)-farnesyl diphosphate mutant enzyme T399S, at pH 7.0 and 30°C Artemisia annua
174
-
 (2E,6E)-farnesyl diphosphate mutant enzyme T447S, at pH 7.0 and 30°C Artemisia annua