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Literature summary extracted from

  • Picaud, S.; Olsson, M.E.; Brodelius, P.E.
    Improved conditions for production of recombinant plant sesquiterpene synthases in Escherichia coli (2007), Protein Expr. Purif., 51, 71-79.
    View publication on PubMed

Application

EC Number Application Comment Organism
4.2.3.77 synthesis improved conditions for expression of Zingiber officinale (+)-germacrene synthase in Escherichia coli. Comparison of bacterial strains; BL21 (DE3), BL21 (DE3) Tuner BL21(DE3) pLysS and BL21 (DE3) pLysS Tuner using different inducing agents. The change between BL21 (DE3) cells and BL21 (DE3) Tuner, induced with IPTG, leads to a twofold increase in enzyme activity in the soluble fraction while a reduction in activity is observed when using the pLysS strains. The same doubling of activity is observed for germacrene synthase when the commonly used BL.21 (DE3) is induced with The Inducer. Addition of 2.5 mM glycine betaine and 660 mM sorbitol to the bacterial growth media results in reduction of growth rate and biomass yield but under these conditions the best overall protein production is obtained Zingiber officinale

Cloned(Commentary)

EC Number Cloned (Comment) Organism
4.2.3.24 into the pET28 vector for expression in Escherichia coli BL21DE3, BL21DE3 Tuner, BL21DE3 pLysS and BL21DE3 pLysS Tuner cells using different inducing agents Artemisia annua
4.2.3.77 expression in Escherichia coli Zingiber officinale

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
4.2.3.24 2-trans,6-trans-farnesyl diphosphate Artemisia annua
-
 amorpha-4,11-diene + diphosphate
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
4.2.3.24 Artemisia annua Q9AR04
-
-
4.2.3.77 Zingiber officinale Q0VHD6
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
4.2.3.24 purified by immobilized metal affinity chromatography on a column loaded with Co2+ Artemisia annua

Renatured (Commentary)

EC Number Renatured (Comment) Organism
4.2.3.24 protein is recovered from the inclusion body fraction Artemisia annua

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
4.2.3.24 0.000002
-
 strain BL21DE3 pLysS Tuner, 6 g/l Inducer, sorbitol, betaine Artemisia annua
4.2.3.24 0.000002
-
 strain BL21DE3 pLysS Tuner, no Inducer Artemisia annua
4.2.3.24 0.000002
-
 strain BL21DE3 pLysS, without IPTG, sorbitol, betaine Artemisia annua
4.2.3.24 0.000003
-
 strain BL21DE3 pLysS Tuner, 1 g/l Inducer, sorbitol, betaine Artemisia annua
4.2.3.24 0.000003
-
 strain BL21DE3 pLysS Tuner, 3 g/l Inducer, sorbitol, betaine Artemisia annua
4.2.3.24 0.000003
-
 strain BL21DE3 pLysS Tuner, no Inducer, sorbitol, betaine Artemisia annua
4.2.3.24 0.000008
-
 strain BL21DE3 pLysS Tuner, 6 g/l Inducer Artemisia annua
4.2.3.24 0.000008
-
 strain BL21DE3 pLysS Tuner, without IPTG Artemisia annua
4.2.3.24 0.000008
-
 strain BL21DE3 pLysS Tuner, without IPTG, sorbitol, betaine Artemisia annua
4.2.3.24 0.000013
-
 strain BL21DE3 pLysS Tuner, 1 g/l Inducer Artemisia annua
4.2.3.24 0.000013
-
 strain BL21DE3 pLysS Tuner, 3 g/l Inducer Artemisia annua
4.2.3.24 0.00002
-
 strain BL21DE3 pLysS Tuner, 0.1 mM IPTG Artemisia annua
4.2.3.24 0.00002
-
 strain BL21DE3 pLysS Tuner, 0.2 mM IPTG Artemisia annua
4.2.3.24 0.00002
-
 strain BL21DE3 pLysS Tuner, 0.5 mM IPTG Artemisia annua
4.2.3.24 0.00002
-
 strain BL21DE3 pLysS, 6 g/l Inducer Artemisia annua
4.2.3.24 0.00002
-
 strain BL21DE3 pLysS, no Inducer Artemisia annua
4.2.3.24 0.00002
-
 strain BL21DE3 pLysS, no Inducer, sorbitol, betaine Artemisia annua
4.2.3.24 0.00002
-
 strain BL21DE3 pLysS, without IPTG Artemisia annua
4.2.3.24 0.00003
-
 strain BL21DE3 pLysS, 0.1 mM IPTG Artemisia annua
4.2.3.24 0.00003
-
 strain BL21DE3 pLysS, 0.2 mM IPTG Artemisia annua
4.2.3.24 0.00003
-
 strain BL21DE3 pLysS, 0.5 mM IPTG Artemisia annua
4.2.3.24 0.00003
-
 strain BL21DE3 pLysS, 1 g/l Inducer Artemisia annua
4.2.3.24 0.00003
-
 strain BL21DE3 pLysS, 1 g/l Inducer, sorbitol, betaine Artemisia annua
4.2.3.24 0.00003
-
 strain BL21DE3 pLysS, 3 g/l Inducer Artemisia annua
4.2.3.24 0.00003
-
 strain BL21DE3 pLysS, 3 g/l Inducer, sorbitol, betaine Artemisia annua
4.2.3.24 0.00003
-
 strain BL21DE3 pLysS, 6 g/l Inducer, sorbitol, betaine Artemisia annua
4.2.3.24 0.00003
-
 strain BL21DE3 Tuner, 0.1 mM IPTG Artemisia annua
4.2.3.24 0.00003
-
 strain BL21DE3 Tuner, 0.2 mM IPTG Artemisia annua
4.2.3.24 0.00003
-
 strain BL21DE3 Tuner, 0.5 mM IPTG Artemisia annua
4.2.3.24 0.00003
-
 strain BL21DE3 Tuner, no Inducer Artemisia annua
4.2.3.24 0.00003
-
 strain BL21DE3 Tuner, without IPTG Artemisia annua
4.2.3.24 0.00003
-
 strain BL21DE3, 0.1 mM IPTG Artemisia annua
4.2.3.24 0.00003
-
 strain BL21DE3, 0.2 mM IPTG Artemisia annua
4.2.3.24 0.00003
-
 strain BL21DE3, 0.5 mM IPTG Artemisia annua
4.2.3.24 0.00003
-
 strain BL21DE3, 1 g/l Inducer Artemisia annua
4.2.3.24 0.00003
-
 strain BL21DE3, 3 g/l Inducer Artemisia annua
4.2.3.24 0.00003
-
 strain BL21DE3, 6 g/l Inducer Artemisia annua
4.2.3.24 0.00005
-
 strain BL21DE3 Tuner, 1 g/l Inducer Artemisia annua
4.2.3.24 0.00005
-
 strain BL21DE3 Tuner, 3 g/l Inducer Artemisia annua
4.2.3.24 0.00005
-
 strain BL21DE3 Tuner, 6 g/l Inducer Artemisia annua
4.2.3.24 0.00005
-
 strain BL21DE3, no Inducer Artemisia annua
4.2.3.24 0.00005
-
 strain BL21DE3, without IPTG Artemisia annua
4.2.3.24 0.00006
-
 strain BL21DE3, without IPTG, sorbitol, betaine Artemisia annua
4.2.3.24 0.00008
-
 strain BL21DE3 Tuner, without IPTG, sorbitol, betaine Artemisia annua
4.2.3.24 0.0001
-
 strain BL21DE3 Tuner, 1 g/l Inducer, sorbitol, betaine Artemisia annua
4.2.3.24 0.00012
-
 strain BL21DE3 Tuner, 3 g/l Inducer, sorbitol, betaine Artemisia annua
4.2.3.24 0.00012
-
 strain BL21DE3, 1 g/l Inducer, sorbitol, betaine Artemisia annua
4.2.3.24 0.00012
-
 strain BL21DE3, no Inducer, sorbitol, betaine Artemisia annua
4.2.3.24 0.00013
-
 strain BL21DE3 Tuner, 6 g/l Inducer, sorbitol, betaine Artemisia annua
4.2.3.24 0.00013
-
 strain BL21DE3 Tuner, no Inducer, sorbitol, betaine Artemisia annua
4.2.3.24 0.00013
-
 strain BL21DE3, 3 g/l Inducer, sorbitol, betaine Artemisia annua
4.2.3.24 0.00015
-
 strain BL21DE3, 0.2 mM IPTG, sorbitol, betaine Artemisia annua
4.2.3.24 0.00015
-
 strain BL21DE3, 6 g/l Inducer, sorbitol, betaine Artemisia annua
4.2.3.24 0.0002
-
 strain BL21DE3 pLysS Tuner, 0.1 mM IPTG, sorbitol, betaine Artemisia annua
4.2.3.24 0.0002
-
 strain BL21DE3 pLysS, 0.1 mM IPTG, sorbitol, betaine Artemisia annua
4.2.3.24 0.0002
-
 strain BL21DE3 pLysS, 0.2 mM IPTG, sorbitol, betaine Artemisia annua
4.2.3.24 0.0002
-
 strain BL21DE3 pLysS, 0.5 mM IPTG, sorbitol, betaine Artemisia annua
4.2.3.24 0.0002
-
 strain BL21DE3 Tuner, 0.1 mM IPTG, sorbitol, betaine Artemisia annua
4.2.3.24 0.0002
-
 strain BL21DE3 Tuner, 0.2 mM IPTG, sorbitol, betaine Artemisia annua
4.2.3.24 0.0002
-
 strain BL21DE3 Tuner, 0.5 mM IPTG, sorbitol, betaine Artemisia annua
4.2.3.24 0.0002
-
 strain BL21DE3, 0.1 mM IPTG, sorbitol, betaine Artemisia annua
4.2.3.24 0.0002
-
 strain BL21DE3, 0.5 mM IPTG, sorbitol, betaine Artemisia annua
4.2.3.24 0.00025
-
 strain BL21DE3 pLysS Tuner, 0.2 mM IPTG, sorbitol, betaine Artemisia annua
4.2.3.24 0.00025
-
 strain BL21DE3 pLysS Tuner, 0.5 mM IPTG, sorbitol, betaine Artemisia annua

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4.2.3.24 2-trans,6-trans-farnesyl diphosphate
-
 Artemisia annua amorpha-4,11-diene + diphosphate
-
 ?

Synonyms

EC Number Synonyms Comment Organism
4.2.3.24 ADS
-
 Artemisia annua
4.2.3.24 amorpha-4,11-diene synthase
-
 Artemisia annua

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
4.2.3.24 30
-
 activity assay Artemisia annua

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
4.2.3.24 6.5
-
 activity assay Artemisia annua