An Approach to Increasing Yields of a Sunscreen Compound in Recombinant Yeast Public Deposited

http://ir.library.oregonstate.edu/concern/undergraduate_thesis_or_projects/qr46r2736

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  • The yeast Saccharomyces cerevisiae, commonly used in winemaking, baking, and brewing, also serves as a host for the production of therapeutically valuable pharmaceuticals. Recently, the Bakalinsky laboratory constructed a recombinant yeast strain expressing two zebrafish (Danio rerio) genes in order to produce a UV-protective “sunscreen” compound called gadusol. An antioxidant with sunscreen activity and commercial potential, gadusol is derived from the pentose phosphate pathway (PPP) intermediate sedoheptulose 7-phosphate (S7P) that occurs naturally in yeast. Because it is likely that increased S7P levels can increase gadusol yields in yeast, a method was sought to quantify glucose flux through the PPP relative to flux through glycolysis, the main glucose-consuming pathway in this organism. This thesis project developed such a method and used it to compare glucose flux in the gadusol-producing yeast relative to an isogenic, nongadusol- producing control strain. By feeding cells labeled ¹³C₁-glucose and measuring 𝛿¹³C of CO₂ produced by yeast, using isotope ratio mass spectrometry (IRMS), glucose flux through the PPP was found to be 2.5% higher in the gadusol-producing yeast than in the control strain. This method will provide a simple means of monitoring the effectiveness of cultural and genetic interventions to increase glucose flux through the PPP.
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