Enhanced Glutathione Production in <i>Saccharomyces cerevisiae</i> by High-Throughput Screening System Based on Atmospheric and Room Temperature Plasma (ARTP) Mutagenesis

In this study, we established a mutagenesis and high-throughput screening system to select a high-yielding glutathione (GSH)-producing strain of <i>Saccharomyces cerevisiae</i>. The parent strain was mutated by atmospheric and room temperature plasma (ARTP) technology and cultivated usin...

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Main Authors: Lan Li, Zejian Wang, Ali Mohsin, Yingping Zhuang
Format: Article
Language:English
Published: MDPI AG 2025-04-01
Series:Fermentation
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Online Access:https://www.mdpi.com/2311-5637/11/4/220
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author Lan Li
Zejian Wang
Ali Mohsin
Yingping Zhuang
author_facet Lan Li
Zejian Wang
Ali Mohsin
Yingping Zhuang
author_sort Lan Li
collection DOAJ
description In this study, we established a mutagenesis and high-throughput screening system to select a high-yielding glutathione (GSH)-producing strain of <i>Saccharomyces cerevisiae</i>. The parent strain was mutated by atmospheric and room temperature plasma (ARTP) technology and cultivated using ethionine plate cultivation. Subsequently, high-throughput screening was performed using liquid deep microtiter plates (MTPs) for cultivation and a microplate reader for rapid GSH detection. The results demonstrated the successful selection of a stable mutant strain, S-272, which exhibited significantly enhanced GSH production. Fermentation validation in 5 L bioreactors revealed that S-272 achieved a 14.7% higher final GSH concentration and a 19.5% higher intracellular GSH content compared to the parent strain. The improved performance of S-272 was attributed to enhanced ethanol utilization, elevated activity of γ-glutamylcysteine synthetase (γ-GCS), and increased intracellular trehalose content. This study presents an effective strategy for developing high GSH-yield strains using ARTP complex mutagenesis technology combined with high-throughput screening.
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spelling doaj-art-89cc239977674a8dbe07809cd74c74b72025-08-20T02:28:28ZengMDPI AGFermentation2311-56372025-04-0111422010.3390/fermentation11040220Enhanced Glutathione Production in <i>Saccharomyces cerevisiae</i> by High-Throughput Screening System Based on Atmospheric and Room Temperature Plasma (ARTP) MutagenesisLan Li0Zejian Wang1Ali Mohsin2Yingping Zhuang3State Key Laboratory of Bioreactor Engineering, East China University of Science & Technology, Shanghai 200237, ChinaState Key Laboratory of Bioreactor Engineering, East China University of Science & Technology, Shanghai 200237, ChinaState Key Laboratory of Bioreactor Engineering, East China University of Science & Technology, Shanghai 200237, ChinaState Key Laboratory of Bioreactor Engineering, East China University of Science & Technology, Shanghai 200237, ChinaIn this study, we established a mutagenesis and high-throughput screening system to select a high-yielding glutathione (GSH)-producing strain of <i>Saccharomyces cerevisiae</i>. The parent strain was mutated by atmospheric and room temperature plasma (ARTP) technology and cultivated using ethionine plate cultivation. Subsequently, high-throughput screening was performed using liquid deep microtiter plates (MTPs) for cultivation and a microplate reader for rapid GSH detection. The results demonstrated the successful selection of a stable mutant strain, S-272, which exhibited significantly enhanced GSH production. Fermentation validation in 5 L bioreactors revealed that S-272 achieved a 14.7% higher final GSH concentration and a 19.5% higher intracellular GSH content compared to the parent strain. The improved performance of S-272 was attributed to enhanced ethanol utilization, elevated activity of γ-glutamylcysteine synthetase (γ-GCS), and increased intracellular trehalose content. This study presents an effective strategy for developing high GSH-yield strains using ARTP complex mutagenesis technology combined with high-throughput screening.https://www.mdpi.com/2311-5637/11/4/220glutathione (GSH)<i>Saccharomyces cerevisiae</i>high-throughput screeningatmospheric and room temperature plasma (ARTP)fermentation
spellingShingle Lan Li
Zejian Wang
Ali Mohsin
Yingping Zhuang
Enhanced Glutathione Production in <i>Saccharomyces cerevisiae</i> by High-Throughput Screening System Based on Atmospheric and Room Temperature Plasma (ARTP) Mutagenesis
Fermentation
glutathione (GSH)
<i>Saccharomyces cerevisiae</i>
high-throughput screening
atmospheric and room temperature plasma (ARTP)
fermentation
title Enhanced Glutathione Production in <i>Saccharomyces cerevisiae</i> by High-Throughput Screening System Based on Atmospheric and Room Temperature Plasma (ARTP) Mutagenesis
title_full Enhanced Glutathione Production in <i>Saccharomyces cerevisiae</i> by High-Throughput Screening System Based on Atmospheric and Room Temperature Plasma (ARTP) Mutagenesis
title_fullStr Enhanced Glutathione Production in <i>Saccharomyces cerevisiae</i> by High-Throughput Screening System Based on Atmospheric and Room Temperature Plasma (ARTP) Mutagenesis
title_full_unstemmed Enhanced Glutathione Production in <i>Saccharomyces cerevisiae</i> by High-Throughput Screening System Based on Atmospheric and Room Temperature Plasma (ARTP) Mutagenesis
title_short Enhanced Glutathione Production in <i>Saccharomyces cerevisiae</i> by High-Throughput Screening System Based on Atmospheric and Room Temperature Plasma (ARTP) Mutagenesis
title_sort enhanced glutathione production in i saccharomyces cerevisiae i by high throughput screening system based on atmospheric and room temperature plasma artp mutagenesis
topic glutathione (GSH)
<i>Saccharomyces cerevisiae</i>
high-throughput screening
atmospheric and room temperature plasma (ARTP)
fermentation
url https://www.mdpi.com/2311-5637/11/4/220
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AT zejianwang enhancedglutathioneproductioninisaccharomycescerevisiaeibyhighthroughputscreeningsystembasedonatmosphericandroomtemperatureplasmaartpmutagenesis
AT alimohsin enhancedglutathioneproductioninisaccharomycescerevisiaeibyhighthroughputscreeningsystembasedonatmosphericandroomtemperatureplasmaartpmutagenesis
AT yingpingzhuang enhancedglutathioneproductioninisaccharomycescerevisiaeibyhighthroughputscreeningsystembasedonatmosphericandroomtemperatureplasmaartpmutagenesis