Transcriptional Downregulation of Methanol Metabolism Key Genes During Yeast Death in Engineered Pichia pastoris

IF 3.2 3区 生物学 Q2 BIOCHEMICAL RESEARCH METHODS
Chenbo Wang, Wei Jiang, Chang Yu, Jianye Xia
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Abstract

Pichia pastoris possesses the unique ability to utilize methanol as its sole carbon source, which makes it a proper host for producing various high-value-added products via metabolic engineering. Nevertheless, cell death has been observed during the fermentation of modified P. pastoris, with limited literature elucidating the underlying causes and mechanisms. Understanding the death mechanisms during methanol-based fermentation is crucial for optimizing fermentation strategies, enhancing the accumulation of target products, and reducing production costs. Here, we first sought to eliminate the potential causes of cell death during fermentation, such as inadequate inorganic salts and toxic by-product accumulation. The elimination of these potential causes was achieved efficiently utilizing the high-throughput fermentation equipment. Subsequently, we established a correlation between yeast cell death and the duration of the methanol metabolism period by monitoring the growth of the yeast at different fermentation stages. A critical revelation from this work came from analyzing the yeast's transcriptomic data at various stages of methanol metabolism. It was observed that a significant characteristic of yeast cell death during fermentation was the marked down-regulation of transcript levels of key enzymes involved in the methanol assimilation pathway and genes related to their biosynthesis process. The findings of this work are crucial for better understanding the causes and mechanisms of cell death for engineered P. pastoris during methanol-utilized fermentation.

Abstract Image

酵母死亡过程中甲醇代谢关键基因的转录下调
Pichia pastoris 具有利用甲醇作为唯一碳源的独特能力,这使其成为通过代谢工程生产各种高附加值产品的合适宿主。尽管如此,在改良牧马人发酵过程中仍观察到细胞死亡现象,而阐明其根本原因和机制的文献却很有限。了解甲醇发酵过程中的死亡机制对于优化发酵策略、提高目标产品的积累和降低生产成本至关重要。在这里,我们首先寻求消除发酵过程中细胞死亡的潜在原因,如无机盐不足和有毒副产品积累。我们利用高通量发酵设备有效地消除了这些潜在原因。随后,我们通过监测酵母在不同发酵阶段的生长情况,确定了酵母细胞死亡与甲醇代谢期持续时间之间的相关性。通过分析酵母在甲醇代谢不同阶段的转录组数据,我们得到了一个重要启示。据观察,发酵过程中酵母细胞死亡的一个重要特征是参与甲醇同化途径的关键酶及其生物合成过程相关基因的转录水平明显下调。这项工作的发现对于更好地理解甲醇利用发酵过程中工程酿酒酵母细胞死亡的原因和机制至关重要。
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来源期刊
Biotechnology Journal
Biotechnology Journal Biochemistry, Genetics and Molecular Biology-Molecular Medicine
CiteScore
8.90
自引率
2.10%
发文量
123
审稿时长
1.5 months
期刊介绍: Biotechnology Journal (2019 Journal Citation Reports: 3.543) is fully comprehensive in its scope and publishes strictly peer-reviewed papers covering novel aspects and methods in all areas of biotechnology. Some issues are devoted to a special topic, providing the latest information on the most crucial areas of research and technological advances. In addition to these special issues, the journal welcomes unsolicited submissions for primary research articles, such as Research Articles, Rapid Communications and Biotech Methods. BTJ also welcomes proposals of Review Articles - please send in a brief outline of the article and the senior author''s CV to the editorial office. BTJ promotes a special emphasis on: Systems Biotechnology Synthetic Biology and Metabolic Engineering Nanobiotechnology and Biomaterials Tissue engineering, Regenerative Medicine and Stem cells Gene Editing, Gene therapy and Immunotherapy Omics technologies Industrial Biotechnology, Biopharmaceuticals and Biocatalysis Bioprocess engineering and Downstream processing Plant Biotechnology Biosafety, Biotech Ethics, Science Communication Methods and Advances.
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