Adaptive evolution of engineered yeast for squalene production improvement and its genome‐wide analysis
- 1 March 2021
- Vol. 38 (7), 424-437
- https://doi.org/10.1002/yea.3559
Abstract
In the present study, the adaptive evolution of a metabolically engineered Saccharomyces cerevisiae strain in the presence of an enzyme inhibitor terbinafine for enhanced squalene accumulation via serial transfer leads to the development of robust strains. After adaptation for nearly 1500 h, a strain with higher squalene production efficiency was identified at a specific growth rate of 0.28 h−1 with a final squalene titer of 193 mg/L, which is 16.5‐fold higher than the BY4741 and 3‐fold higher over the metabolically engineered SK22 strain. Whole‐genome sequencing comparison between the reference strain and the evolved variant SK23 has led to the identification of 462 single‐nucleotide variants (SNVs) between both strains, with 102 SNVs affecting metabolism‐related genes. It was also established that F420I mutation of ERG1 in S. cerevisiae improves squalene synthesis. Further, the effect of increased squalene on lipid droplet and neutral lipid pattern in the evolved mutant strains was investigated by fluorescent techniques proving that the neutral lipid content and clustering of lipid droplets increase with an increase in squalene.Keywords
Funding Information
- Karl-Franzens-Universität Graz (KLN1)
- Department of Biotechnology, Government of West Bengal
- Science and Engineering Research Board
- Science and Engineering Research Board (CRG/2019/003323)
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