Abstract
Approaches for improving astaxanthin yields in Xanthophyllomyces dendrorhous include optimization of fermentation conditions and generation of hyperproducing mutants through random mutagenesis using chemical or physical means. A key limitation of classical mutagenesis is the labor-intensive nature of the screening processes required to find relatively rare mutants having increased carotenoid content, as these are present against a high background of low-interest cells. Here, flow cytometry is described as a high-throughput, single-cell method for primary enrichment of mutagenized cells expressing high levels of astaxanthin. This approach improves the speed and productivity of classical strain selection, enhancing the chances for isolating the carotenoid hyperproducing mutants (CHMs) needed to enable high-titer, economical production of natural astaxanthin.
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References
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Acknowledgments
The methods described here are based on the work of Gil-Hwan An, whose doctoral work in the Johnson laboratory was focused on “Improved astaxanthin production from the red yeast P. rhodozyma” (4), and on the work of Ukibe et al. (5) who recently revisited and revised our original protocol for FCCS-based selection of CHMs.
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Brehm-Stecher, B.F., Johnson, E.A. (2012). Isolation of Carotenoid Hyperproducing Mutants of Xanthophyllomyces dendrorhous (Phaffia rhodozyma) by Flow Cytometry and Cell Sorting. In: Barredo, JL. (eds) Microbial Carotenoids From Fungi. Methods in Molecular Biology, vol 898. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-918-1_14
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DOI: https://doi.org/10.1007/978-1-61779-918-1_14
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