Part:BBa_K5184003
ShZPO
To equip our insecticide with enhanced prevention efficacy against T. urticae, we also decide to synthesize 9-hydroxy-zingiberene (9HZ) and 9-hydroxy-10,11-epoxy zingiberene (9H10epoZ), two oxidized products of the monocyclic sesquiterpene 7epiZ. This requires the involvement of ShZPO, a zingiberene oxidase, which is able to produce 9HZ and 9H10epoZ from 7epiZ in E. coli. It will be collectively used with the reductase SlCPR2 or AtCPR1, which acts as its redox partner and electron supplier. Our exploration of the oxidase provide future iGEM team with a novel way of generating sesquiterpenes from a monocyclic sesquiterpene through oxidation carried out by the collaboration of an oxidase and a reductase.
Usage and Biology
ShZPO is a cytochrome P450 oxygenase found in Solanum habrochaites. It carries out two successive oxidations to generate two sesquiterpenes from a monocyclic sesquiterpene as the susbtrate. ShZPO requires a cytovhrome P450 reductase, in our case SlCPR2 or AtCPR1 as its redox partner and electron supplier. The enzyme was found to be predominantly localized on the ER membrane. The co-localization of oxidase ShZPO and reductase SlCPR2 on the ER membrane ensures efficient electron transfer. We constructed a novel sesquiterpene synthesis pathway in E. coli. Using glucose as our raw material, we introduce the MVA pathway, which transforms glucose into dimethylallyl diphosphate (DMAPP) and isopentenyl diphosphate (IPP). Afterwards, SltNPPS, a neryl diphosphate synthase catalyze the production of NPP from IPP and DMAPP. Mvan4662 is then introduced to catalyze the formation of Z,Z-FPP. Then, ShZIS transforms Z,Z-FPP into 7epiZ. In the end, ShZPO works collaboratively with SlCPR2 or AtCPR1.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
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