Team:British Columbia/Notebook/Week 20

Diterpene Production in Yeast with LAS and ISO diterpene synthases
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.

G. clavigera and S. cerevisiae co-culture experiments

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

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

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

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.

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

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