Team:British Columbia/Notebook/Week 20

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Team: British Columbia - 2011.igem.org

Week 20: Oct 16-22



Diterpene production in yeast with LAS and ISO


We obtained four yeast strains from Katrin and Niels (from Bohlmann lab). The yeast strains contained

  1. Levoabietadiene synthase + CYP720B4
  2. Levoabietadiene synthase
  3. Isopimaradiene synthase + CYP720B4
  4. Isopimaradiene synthase

These are diterpene synthases which convert GGPP to the diterpinols, which are hydrolyzed to form diterpenes. These diterpenes can be further hydroxylated to form resin acids. CYP720B4 is an enzyme isolated by members of Bohlmann lab. This enzyme plays a key role in the production of specific acids that have been shown to have anti-white pine weevil effects.



Ubcigem2011beetleplatefungi.jpg

Ubcigem2011beetleplatefungi1.jpg

Ubcigem2011beetleplatefungi2.jpg

Ubcigem2011beetleplatefungi3.jpg

G. clavigera and S. cerevisiae co-culture experiments

Cocultureubcigem2011.jpg

200 G. clavigera spores plated on 1% OMEA for 4 days.


2cocultureubcigem2011.jpg

330 S. cerevisiae spores plated on 1% OMEA for 4 days.


1cocultureubcigem2011.jpg

200 G. clavigera + 330 S. cerevisiae spores + plated on 1% OMEA for 4 days.


3cocultureubcigem2011.jpg

Assorted plates of G. clavigera mycelium plugs and terpenoid-producing S. cerevisiae rings.

S. cerevisiae grows well on YPD media but the G.claviegera grows very slowly.

S. cerevisiae grows slower on 1% OMEA but the G.clavigera grows well.


4cocultureubcigem2011.jpg

G. clavigera mycelium growing outwards towards diterpenoid ISO-producing S. cerevisiae on 1% OMEA.


5cocultureubcigem2011.jpg

G. clavigera mycelium growing very slowly outwards towards diterpenoid P40-producing S. cerevisiae on YPD.


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