Part:BBa_K5184018
SltNPPS-Mvan4662-ShZIS-st-t25ShZPO-sc-t76SlCPR2
SltNPPS (part number), an neryl diphosphate synthase with its N-termial 45 amino acid-trucated. Originally found in Solanum Lycopersium, it is used for transformation of glucose into IPP and DMAPP. Mvan4662 (part number), a farnesyl diphosphate synthase facilitating the formation of longer-chain isoprenoid diphosphates. In our context, it is included for synthesis of Z,Z-FPP from NPP. ShZIS, a zingiberene synthase derived from Solanum habrochaites, catalyzes the formation of sesquiterpenes, a compound with 15 carbons. Specifically, we us ShZIS to produce 7-epi-zingiberene from its substrate Z,Z-FPP. ShZPO, a cytochrome P450 oxygenase found in Solanum habrochaites, carries out two successive oxidations to generate two sesquiterpenes from a monocyclic sesquiterpene as the susbtrate. In our context, ShZPO will act as a zingiberene oxidase, working with its redox partner SlCPR2, a NADPH-cytochrome P450 reductase found in Solanum lycopersicum, to produce the zingiberenoids 9-hydroxy-zingiberene and 9-hydroxy-10,11-epoxyzingiberene from 7-epi-zingiberene. In addition, a spytag-spycatcher system was incorporated, with spytag linked to ShZPO forming an isopeptide bond with the spycathcer linked to SlCPR2. This allows more efficient electron transfer between the two cytochrome P450 enzymes through imitating the colocalization of them on the ER membrane in eukaryotic plants.
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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