Fatty acid requiring S. cerevisiae strains such as fas1- or fas2- needs myristic acid and Tween in the medium to be able to grow.
- 0.5% - 1.0% (v/v) Tween 40 (PubChem)
- 1.3 - 2 mM myristic acid M
w= 228.37 g/mol (C14) (hmy) (PubChem)
Tween 40 is the commercial name for Polyoxyethylene (20) sorbitan monopalmitate. This compound provides provides palmitic acid (C16) and helps solubilize the myristic acid.
Preparation:
- Heat the bottle containing Tween 40 briefly in the microwave, 15 s.
- Add Tween 40 to 0.5% or 1/200 of the final medium volume.
- Add 2 mM = 0.460 g/L myristic acid (powder).
- 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.