Screening of astaxanthin-overproducing Xanthophyllomyces dendrorhous strains via iterative ARTP mutagenesis and cell sorting by flow cytometry

Author:

Ding Ruirui123,Huang Ruilin1234,Su Hang1234,Li Jiawen123,Li Fuli123,Wang Shi'an123ORCID

Affiliation:

1. Key Laboratory of Biofuels, Qingdao Carbon One Biorefining Research Center, Qingdao Institute of Bioenergy and Bioprocess Technology, Chinese Academy of Sciences , Qingdao 266100 , China

2. Shandong Energy Institute , Qingdao 266100 , China

3. Qingdao New Energy Shandong Laboratory , Qingdao 266100 , China

4. University of Chinese Academy of Sciences , College of Life Sciences, Beijing, 100039 , China

Abstract

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 g−1 DCW (dry cell weight) and 5.30 mg astaxanthin g−1 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.

Funder

Natural Key R&D Program of China

Shandong Energy Institute

Publisher

Oxford University Press (OUP)

Reference35 articles.

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