1. Screening of astaxanthin-overproducing Xanthophyllomyces dendrorhous strains via iterative ARTP mutagenesis and cell sorting by flow cytometry.
- Author
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Ding, Ruirui, Huang, Ruilin, Su, Hang, Li, Jiawen, Li, Fuli, and Wang, Shi'an
- Subjects
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ASTAXANTHIN , *FLOW cytometry , *MUTAGENESIS - Abstract
Aims The astaxanthin-producing yeast Xanthophyllomyces dendrorhous is widely used in aquaculture. Due to the production of carotenoid, this yeast shows visible color; however, high-throughput approaches for identification of astaxanthin-overproducing strains remain rare. Methods and results This study verified an effective approach to identify astaxanthin-overproducing mutants of X. dendrorhous by flow cytometry (FCM) and cell sorting. First, the mutant libraries were generated by atmospheric and room-temperature plasma (ARTP) mutagenesis. Second, a highly direct correlation between the concentrations of intracellular astaxanthin and the levels of emitting fluorescence was constructed by testing a variety of astaxanthin-contained populations via FCM and cell sorting. Third, iterative cell sorting efficiently improves the identification of astaxanthin-overproducing strains. Finally, two mutants producing 4.96 mg astaxanthin gā1 DCW (dry cell weight) and 5.30 mg astaxanthin gā1 DCW were obtained, which were 25.3% and 33.8% higher than that of the original strain, respectively. Conclusions This study demonstrated that iterative ARTP mutagenesis along with cell sorting by FCM is effective for identifying astaxanthin-overproduction strains. [ABSTRACT FROM AUTHOR]
- Published
- 2024
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