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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
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 confer resistance to the white pine weevil, part of the reason why the iGEM team decided to characterize this part in yeast. 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.
The 3-carene synthase has been previously characterized by expression in C41 DE3 E. coli followed by a His-tag purification. The purified enzymes were assayed in vitro with GPP and sent for GC-MS.
The iGEM team aims to characterize the 3-carene synthase in vivo in S. cerevisiae yeast. The products wil be sent for GC-MS.
Reference: 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 March 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
