Cloned (Comment) | Organism |
---|---|
mutated enzymes are generated using the QuickChange II site-directed mutagenesis kit. The PCR-based mutagenesis protocol is performed using pET200-D-TOPO harbouring the respective AcTPS2 cDNAs as template. The single-site mutant enzymes overexpressed in Escherichia coli. | Actinidia chinensis |
Protein Variants | Comment | Organism |
---|---|---|
K514A | AcTPS2 mutant, 90% reduction in monoterpene synthase activity compared to wild-type enzyme | Actinidia chinensis |
K514S | AcTPS2 mutant, presence of Ser is not able to rescue any of the activity lost after the removal of the Lys residue | Actinidia chinensis |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | 10 mM are included in farnesyl diphosphate activity assay | Actinidia chinensis | |
additional information | wild-type AcTPS2 is significantly more active when KCl is absent from the activity buffer | Actinidia chinensis |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | Actinidia chinensis | terpenes produced from the wild-type and mutant enzymes are identical, showing low levels of beta-myrcene from AcTPS2-K514A | ? | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Actinidia chinensis | - |
- |
- |
Purification (Comment) | Organism |
---|---|
recombinant AcTPS2 protein is extracted and purified | Actinidia chinensis |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | terpenes produced from the wild-type and mutant enzymes are identical, showing low levels of beta-myrcene from AcTPS2-K514A | Actinidia chinensis | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
AcTPS2 | - |
Actinidia chinensis |
myrcene synthase | - |
Actinidia chinensis |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
farnesyl diphosphate and geranyl diphosphate activity assay | Actinidia chinensis |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
farnesyl diphosphate activity assay | Actinidia chinensis |