Augmented peroxisomal ROS buffering capacity renders oxidative and thermal stress cross-tolerance in yeast

Lin, Nai-Xin; He, Rui-Zhen; Xu, Yan; Xiang, Yu

doi:10.1186/s12934-021-01623-1

Cited by 7 publications

(7 citation statements)

References 66 publications

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“…Previously, we isolated a robust P. pastoris mutant, which exhibited thermal and oxidative co-stress tolerance. In addition, the robust mutant expressed an up to 2.5-fold higher level of heterologous protein than the WT [19]. To determine whether this phenomenon could also occur in the expression of other proteins, a green fluorescence protein (GFP) was expressed in the robust mutant and its WT.…”

Section: Improved Heterologous Protein Expression Ability In the Robust Mutant Under Methanol Inductionmentioning

confidence: 99%

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Oxidative stress tolerance contributes to heterologous protein production in Pichia pastoris

Lin

et al. 2021

Biotechnol Biofuels

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Background Pichia pastoris (syn. Komagataella phaffii) is an important yeast system for heterologous protein expression. A robust P. pastoris mutant with oxidative and thermal stress cross-tolerance was acquired in our previous study. The robust mutant can express a 2.5-fold higher level of lipase than its wild type (WT) under methanol induction conditions. Results In this study, we found that the robust mutant not only can express a high level of lipase, but also can express a high level of other heterogeneous proteins (e.g., green fluorescence protein) under methanol induction conditions. Additionally, the intracellular reactive oxygen species (ROS) levels in the robust mutant were lower than that in the WT under methanol induction conditions. To figure out the difference of cellular response to methanol between the WT and the robust mutant, RNA-seq was detected and compared. The results of RNA-seq showed that the expression levels of genes related to antioxidant, MAPK pathway, ergosterol synthesis pathway, transcription factors, and the peroxisome pathway were upregulated in the robust mutant compared to the WT. The upregulation of these key pathways can improve the oxidative stress tolerance of strains and efficiently eliminate cellular ROS. Hence, we inferred that the high heterologous protein expression efficiency in the robust mutant may be due to its enhanced oxidative stress tolerance. Promisingly, we have indeed increased the expression level of lipase up to 1.6-fold by overexpressing antioxidant genes in P. pastoris. Conclusions This study demonstrated the impact of methanol on the expression levels of genes in P. pastoris and emphasized the contribution of oxidative stress tolerance on heterologous protein expression in P. pastoris. Our results shed light on the understanding of protein expression mechanism in P. pastoris and provided an idea for the rational construction of robust yeast with high expression ability.

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Section: Improved Heterologous Protein Expression Ability In the Robust Mutant Under Methanol Inductionmentioning

confidence: 99%

“…The robust mutant was previously evolved by adaptive laboratory evolution [19]. Both the robust mutant and its WT expressed a recombinant protein lipase encoded by RCL gene and tightly regulated by the methanolinducible promoter P AOX1 .…”

Section: Strain and Plasmid Constructionmentioning

confidence: 99%

Oxidative stress tolerance contributes to heterologous protein production in Pichia pastoris

Lin

et al. 2021

Biotechnol Biofuels

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“…Adaptive evolution‐derived mutant (G14) of Komagataella phaffii was found to be thermotolerant with reduced lipid peroxidation and ROS levels, owing to bigger peroxisomes in G14 and thus, presenting the enhanced oxidative stress response. In addition, augmented activity and expression of peroxisomal catalase was also observed in the G14 (Lin et al, 2021).…”

Section: Evolutionary Basis For Multiple Stress Conditionsmentioning

confidence: 93%

Recent advances in yeast genome evolution with stress tolerance for green biological manufacturing

Zhang

Guo

et al. 2022

Biotech & Bioengineering

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Green biological manufacturing is a revolutionary industrial model utilizing yeast as a significant microbial cell factory to produce biofuels and other biochemicals. However, biotransformation efficiency is often limited owing to several stress factors resulting from environmental changes or metabolic imbalance, leading to the slow growth of cells, compromised yield, and enhanced energy consumption. These factors make biological manufacturing competitively less economical. In this regard, minimizing the stress impact on microbial cell factories and strong robust performance have been an interesting area of interest in the last few decades. In this review, we focused on revealing the stress factors and their associated mechanisms for yeast in biological manufacturing. To improve yeast tolerance, rational and irrational strategies were introduced, and the molecular basis of genome evolution in yeast was also summarized. Furthermore, strategies of genome‐directed evolution such as homology directed repair and nonhomologous end‐joining, and the synthetic chromosome recombination and modification by LoxP‐mediated evolution and their association with stress tolerance was highlighted. We hope that genome evolution provides new insights for solving the limitations of the natural phenotypes of microorganisms in industrial fermentation for the production of valuable compounds.

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“…Interestingly, the phenomenon that the inner cell wall thickens happened in a lager yeast which exhibited a multiple stress tolerance [74]. Fascinatingly, in our research on P. pastoris, it was also found that the environmental stress tolerance was increased, and the inner cell wall became thicker [75]. Interestingly, the CWI pathway is not the only way to maintain cell wall integrity.…”

Section: Cwi Pathwaymentioning

confidence: 71%

“…This process is achieved mainly by trehalose, UPS, and autophagy antioxidants to scavenge cellular ROS and eliminate oxidative stress. Especially, in our previous work, we found that high temperature induced OSR and HSR simultaneously in a heat-sensitive Pichia pastoris strain [39].…”

Section: Intracellular Accumulation Of Denatured Proteins and Reactive Oxygen Species (Ros) That Can Be Induced By Most Environmental Strmentioning

confidence: 81%

Overview of yeast environmental stress response pathways and the development of tolerant yeasts

Lin

2021

Syst Microbiol and Biomanuf

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Yeast is widely used for industrial production of various types of products, such as ethanol and enzymes. However, its fermentation efficiency is strongly reduced by harmful environmental stresses. Specifically, harmful environmental stresses damage important cellular components, such as cell wall, cell membrane, proteins, etc. Then, these damages cause cellular metabolic disorders or even death. In the past decades, there has been a portfolio of studies on the environmental stress tolerance of yeasts, which mainly aimed at cell damages caused by different environmental stresses, different ways to improve yeast environmental stress tolerance or a tolerance mechanism for certain environmental stress. However, a comprehensive overview of how yeasts respond to environmental stresses is lacking, and the correlation of tolerance mechanism between different environmental stresses is unclear. In this review, we summarized the general damages induced by most of environmental stresses, the existing major mechanisms of environmental stress tolerance from the perspective of key signalling pathways, and the common ways to improve the resistance to environmental stresses in yeast cells. The tolerance mechanisms of yeast cells to different environmental stresses are diverse, but sometimes they share the same signalling pathway. Cells use sensors on the cell surface to recognize environmental stresses and transmit signals to the nucleus to cause changes in gene expression. By summarizing the main signalling pathways, including MAPK pathway, cAMP/PKA pathway, YAP1/ SKN7 pathway, it will provide a powerful reference for future efforts to promote yeast environmental stress tolerance and study yeast tolerance mechanisms.

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Augmented peroxisomal ROS buffering capacity renders oxidative and thermal stress cross-tolerance in yeast

Cited by 7 publications

References 66 publications

Oxidative stress tolerance contributes to heterologous protein production in Pichia pastoris

Oxidative stress tolerance contributes to heterologous protein production in Pichia pastoris

Recent advances in yeast genome evolution with stress tolerance for green biological manufacturing

Overview of yeast environmental stress response pathways and the development of tolerant yeasts

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