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High-Throughput Screening and Dual Feeding Fed-Batch Strategy for Enhanced Single-Cell Oil Accumulation in Yarrowia lipolytica

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Abstract

Herein, we combined metabolic evolution with fluorescence-activated cell sorting (FACS) of cells stained with the lipophilic dye BODIPY for isolation of SCO-overproducing strains of Yarrowia lipolytica. Metabolic evolution was implemented for enrichment of high SCO-accumulating mutant population which were then sorted by fluorescence signals using flow cytometry coupled with FACS. A mutant isolated by this approach exhibited 1.5- and 1.2-fold higher SCO titer and content, respectively, than the wild type under batch culture of sugarcane bagasse hydrolysate complex media. In addition, the mutant had whole-cell fatty acid composition different from that of the wild type with higher oleic and linoleic acids. Dual-stage fed-batch process applied to the mutant yielded high SCO titer of 49.7 g/L from hydrolysates, a fourfold improvement over batch process. This study highlights evolution-based in conjunction with fluorescence-based high-throughput screening as a powerful strategy for attaining high single-cell oil-accumulating phenotype in Y. lipolytica exploited for sustainable biodiesel and oleochemicals synthesis.

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Acknowledgements

The authors would like to thank the National Center for Genetic Engineering and Biotechnology, Thailand, for the financial support of this project (Grant No. P-16-50341).

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  1. National Center for Genetic Engineering and Biotechnology (BIOTEC), 113 Thailand Science Park, Phaholyothin Road, Khlong 1, Khlong Luang, Pathumthani, 12120, Thailand

    Pornkamol Unrean & Verawat Champreda

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  1. Pornkamol Unrean
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  2. Verawat Champreda
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Correspondence to Pornkamol Unrean.

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Unrean, P., Champreda, V. High-Throughput Screening and Dual Feeding Fed-Batch Strategy for Enhanced Single-Cell Oil Accumulation in Yarrowia lipolytica . Bioenerg. Res. 10, 1057–1065 (2017). https://doi.org/10.1007/s12155-017-9865-0

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