Breeding of a High-Yield Glutathione-Producing Strain by Atmospheric and Room Temperature Plasma Mutagenesis and Microbial Microdroplet Culture System
In order to improve the ability of a wild-type strain (BY-1) of Pichia pastoris to produce glutathione (GSH), a mutant strain designated BY-1-26 was obtained by atmospheric and room temperature plasma (ARTP) mutagenesis. To further improve the production of glutathione by the mutant during shake fla...
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| Format: | Article |
| Language: | English |
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China Food Publishing Company
2023-02-01
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| Series: | Shipin Kexue |
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| Online Access: | https://www.spkx.net.cn/fileup/1002-6630/PDF/2023-44-4-027.pdf |
| _version_ | 1827998324884504576 |
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| author | LING Siyu, WANG Zhou, ZHANG Huimin, LI Chuang, LIU Qingtao, LIU Yan, XUE Zhenglian |
| author_facet | LING Siyu, WANG Zhou, ZHANG Huimin, LI Chuang, LIU Qingtao, LIU Yan, XUE Zhenglian |
| author_sort | LING Siyu, WANG Zhou, ZHANG Huimin, LI Chuang, LIU Qingtao, LIU Yan, XUE Zhenglian |
| collection | DOAJ |
| description | In order to improve the ability of a wild-type strain (BY-1) of Pichia pastoris to produce glutathione (GSH), a mutant strain designated BY-1-26 was obtained by atmospheric and room temperature plasma (ARTP) mutagenesis. To further improve the production of glutathione by the mutant during shake flask culture, 1,2,4-triazole was added to the culture medium. Finally, adaptive evolution of the mutant strain was carried out on a microbial microdroplet culture system (MMC) using 1,2,4-triazole as a screening factor. As a result, a high-yield glutathione-producing mutant strain BY-2-24 was obtained, and its genetic stability was investigated. The results showed that a high-yield glutathione-producing mutant strain could be obtained using the wild-type strain BY-1 by ARTP mutagenesis, primary resistance screening on gradient plates, adaptive evolution on MMC and shake-flask secondary screening. The yield of glutathione produced by BY-2-24 in shake flask culture was (312.13 ± 2.62) mg/L, which was 134.26% higher than that produced by the original strain, and this mutant still had good genetic stability after seven passages. Meanwhile, the biomass was increased by 118.33%, indicating that the growth ability of the mutant strain was improved compared to the original strain. The result of this study show that the combination of ARTP and MMC can be used as a simple and effective screening method for excellent mutant strains, which provides a reference for high throughput selection of target strains. |
| first_indexed | 2024-04-10T05:41:40Z |
| format | Article |
| id | doaj.art-0436c01669334fd29c6d4cc9095b3ea7 |
| institution | Directory Open Access Journal |
| issn | 1002-6630 |
| language | English |
| last_indexed | 2024-04-10T05:41:40Z |
| publishDate | 2023-02-01 |
| publisher | China Food Publishing Company |
| record_format | Article |
| series | Shipin Kexue |
| spelling | doaj.art-0436c01669334fd29c6d4cc9095b3ea72023-03-06T07:13:33ZengChina Food Publishing CompanyShipin Kexue1002-66302023-02-0144420020810.7506/spkx1002-6630-20220302-040Breeding of a High-Yield Glutathione-Producing Strain by Atmospheric and Room Temperature Plasma Mutagenesis and Microbial Microdroplet Culture SystemLING Siyu, WANG Zhou, ZHANG Huimin, LI Chuang, LIU Qingtao, LIU Yan, XUE Zhenglian0(Anhui Engineering Laboratory for Industrial Microbiology Molecular Breeding, Anhui Engineering Technology Research Center of Microbial Fermentation, College of Biology and Food Engineering, Anhui Polytechnic University, Wuhu 241000, China)In order to improve the ability of a wild-type strain (BY-1) of Pichia pastoris to produce glutathione (GSH), a mutant strain designated BY-1-26 was obtained by atmospheric and room temperature plasma (ARTP) mutagenesis. To further improve the production of glutathione by the mutant during shake flask culture, 1,2,4-triazole was added to the culture medium. Finally, adaptive evolution of the mutant strain was carried out on a microbial microdroplet culture system (MMC) using 1,2,4-triazole as a screening factor. As a result, a high-yield glutathione-producing mutant strain BY-2-24 was obtained, and its genetic stability was investigated. The results showed that a high-yield glutathione-producing mutant strain could be obtained using the wild-type strain BY-1 by ARTP mutagenesis, primary resistance screening on gradient plates, adaptive evolution on MMC and shake-flask secondary screening. The yield of glutathione produced by BY-2-24 in shake flask culture was (312.13 ± 2.62) mg/L, which was 134.26% higher than that produced by the original strain, and this mutant still had good genetic stability after seven passages. Meanwhile, the biomass was increased by 118.33%, indicating that the growth ability of the mutant strain was improved compared to the original strain. The result of this study show that the combination of ARTP and MMC can be used as a simple and effective screening method for excellent mutant strains, which provides a reference for high throughput selection of target strains.https://www.spkx.net.cn/fileup/1002-6630/PDF/2023-44-4-027.pdfglutathione; pichia pastoris; atmospheric and room temperature plasma; adaptive evolution; strain selection |
| spellingShingle | LING Siyu, WANG Zhou, ZHANG Huimin, LI Chuang, LIU Qingtao, LIU Yan, XUE Zhenglian Breeding of a High-Yield Glutathione-Producing Strain by Atmospheric and Room Temperature Plasma Mutagenesis and Microbial Microdroplet Culture System Shipin Kexue glutathione; pichia pastoris; atmospheric and room temperature plasma; adaptive evolution; strain selection |
| title | Breeding of a High-Yield Glutathione-Producing Strain by Atmospheric and Room Temperature Plasma Mutagenesis and Microbial Microdroplet Culture System |
| title_full | Breeding of a High-Yield Glutathione-Producing Strain by Atmospheric and Room Temperature Plasma Mutagenesis and Microbial Microdroplet Culture System |
| title_fullStr | Breeding of a High-Yield Glutathione-Producing Strain by Atmospheric and Room Temperature Plasma Mutagenesis and Microbial Microdroplet Culture System |
| title_full_unstemmed | Breeding of a High-Yield Glutathione-Producing Strain by Atmospheric and Room Temperature Plasma Mutagenesis and Microbial Microdroplet Culture System |
| title_short | Breeding of a High-Yield Glutathione-Producing Strain by Atmospheric and Room Temperature Plasma Mutagenesis and Microbial Microdroplet Culture System |
| title_sort | breeding of a high yield glutathione producing strain by atmospheric and room temperature plasma mutagenesis and microbial microdroplet culture system |
| topic | glutathione; pichia pastoris; atmospheric and room temperature plasma; adaptive evolution; strain selection |
| url | https://www.spkx.net.cn/fileup/1002-6630/PDF/2023-44-4-027.pdf |
| work_keys_str_mv | AT lingsiyuwangzhouzhanghuiminlichuangliuqingtaoliuyanxuezhenglian breedingofahighyieldglutathioneproducingstrainbyatmosphericandroomtemperatureplasmamutagenesisandmicrobialmicrodropletculturesystem |