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Team:British Columbia/Characterized
From 2011.igem.org
Here are the new parts we aim to characterize:
A variety of monoterpene synthases that produce alpha- and beta- pinene, carene and cineole.
ERG20 and erg20-2
IDI1
HMG20
GAL galactose-inducible yeast promoter
GPD constitutive yeast promoter
3-Carene Synthase
NCBI: HQ426167
The 3-carene synthase is a monoterpene synthase isolated from Picea sitchensis by Chris Keeling and Dr. Joerg Bohlmann. This 3-carene synthase was found to be resistant to white pine weevil, part of the reason why the iGEM team decided to characterize this part in yeast (2011 Keeling). This synthase converts farnesyl pyrophosphate to 66.4% (+)-3-Carene, 16.3% Terpinolene, 2.7% (-)-alpha-pinene, 2.5% Terpinen-4-ol, 2.1% (-)-beta-phellandrene, 2.1% myrcene, 1.4% alpha-Terpinol, 0.8% gamma-terpinene, 1% others (2011 Keeling).
The 3-carene synthase has been previously characterized by expression in C41 DE3 cells, with a His-tag in vitro purification. The purified enzymes were assayed with GPP and sent for GCMS (2011 Keeling).
The iGEM team aims to characterize the 3-carene synthase in vivo in sacchromeyces cerevisae. The products wil be sent for GCMS.
Sources: (1) Keeling, CI, Weisshar S, Ralph SG, Jancsik S, Hamberger B, Dullat HK, and Joerg Bohlmann. Transcriptome mining, functional characterization, and phylogeny of a large terpene synthase gene family in spruce (Picea spp.) BMC. 2011 Mar 7 11: 43.
Here are the existing parts we aim to characterize/improve:
BBa_K118025: Limonene Synthase Composite + Lac promotor BBa_K118024: Limonene Synthase Composite BBa_J63006: Yeast GAL1 Promoter
