Fatty acid requiring S. cerevisiae strains such as fas1- or fas2- needs myristic acid and Tween in the medium to be able to grow.
Tween 40 is the commercial name for Polyoxyethylene (20) sorbitan monopalmitate. This compound provides provides palmitic acid (C16).
Preparation:
- Heat the bottle containing Tween 40 briefly in the microwave, 15 s.
- Add Tween 40 to 1/200 of the final medium volume. For example, pipette 1 mL for 200 mL medium.
- Add 0.460 g/L myristic acid (powder). For example 92 mg to 200 mL.
- Autoclave the medium.
Calculations:
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Literature Support for the Protocol
“For the FAS2 knockout strain, the medium contained 2 mM myristic acid with tween 40 (5 g/L) to supplement growth (Chirala et al., 1987). The fatty acid supplement was heated to 55°C for 30 min to increase solubility and then filter-sterilized.”
Leber, Christopher, and Nancy A Da Silva. 2014. “Engineering of Saccharomyces Cerevisiae for the Synthesis of Short Chain Fatty Acids.” Biotechnol. Bioeng. 111 (2): 347–58. https://doi.org/10.1002/bit.25021.
“To enable growth of fatty acid auxotrophic strains, YPD medium was supplemented with 0.03% myristic acid and 1% Tween 40”
Scazzari, Mario, Ingo Amm, and Dieter H. Wolf. 2015. “Quality Control of a Cytoplasmic Protein Complex: CHAPERONE MOTORS AND THE UBIQUITIN-PROTEASOME SYSTEM GOVERN THE FATE OF ORPHAN FATTY ACID SYNTHASE SUBUNIT Fas2 OF YEAST*.” Journal of Biological Chemistry 290 (8): 4677–87. https://doi.org/10.1074/jbc.M114.596064.