Abstract
Sustainable production of oleochemicals requires establishment of cell factory platform strains. The yeast Saccharomyces cerevisiae is an attractive cell factory as new strains can be rapidly implemented into existing infrastructures such as bioethanol production plants. Here we show high-level production of free fatty acids (FFAs) in a yeast cell factory, and the production of alkanes and fatty alcohols from its descendants. The engineered strain produces up to 10.4 g l−1 of FFAs, which is the highest reported titre to date. Furthermore, through screening of specific pathway enzymes, endogenous alcohol dehydrogenases and aldehyde reductases, we reconstruct efficient pathways for conversion of fatty acids to alkanes (0.8 mg l−1) and fatty alcohols (1.5 g l−1), to our knowledge the highest titres reported in S. cerevisiae. This should facilitate the construction of yeast cell factories for production of fatty acids derived products and even aldehyde-derived chemicals of high value.
Highlights
Sustainable production of oleochemicals requires establishment of cell factory platform strains
We demonstrated for the first time the significant conversion of free fatty acids (FFAs) to alkanes and fatty alcohols in yeast, and we showed that this FFA dependent pathway is far more efficient than the earlier reported route from fatty acyl-CoA (Fig. 3b and Supplementary Fig. 8)
The final engineered strain had a 20% lower biomass level in YPD medium, which indicated that the combination of disrupting FFA activation and neutral lipid recycle was harmful to the cell, and might retard growth further in minimal media with lower and less diverse nutrient availability
Summary
Sustainable production of oleochemicals requires establishment of cell factory platform strains. Plasmid-expression retarded the cell growth probably due to the metabolic burden (Supplementary Fig. 1), we genomicintegrated the optimized actyl-CoA pathway consisting of MmACL, RtME, CTP1 and ‘MDH3, which improved FFA production to 0.80 g l À 1 (strain YJZ41, Fig. 2a). Genomic integration of both RtFAS and the acetyl-CoA pathway (YJZ45) increased the FFA titre to 0.92 g l À 1 in shake flasks and the corresponding prototrophic strain YJZ45U reached 7.0 g l À 1 in fed-batch cultivation.
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
