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<p><i>Pseudomonas plecoglossicida</i> is an industrial strain for the production of 2-keto-D-gluconic acid (2-KGA); however, its fermentation performance and 2-KGA yield significantly decline under high-temperature conditions. Tandem mass tag (TMT)-based quantitative proteomics was performed to investigate thermal adaptation mechanisms in <i>Pseudomonas plecoglossicida</i> at 32&#xa0;°C, 36&#xa0;°C, and 40&#xa0;°C. In total, 565 and 1,665 differentially expressed proteins (DEPs) were identified at 36&#xa0;°C and 40&#xa0;°C vs. 32&#xa0;°C, respectively. Functional enrichment revealed significant proteomic rewiring in carbohydrate metabolism, oxidative phosphorylation, and ubiquinone biosynthesis under thermal stress. The Entner-Doudoroff (ED) pathway acts as a temperature-dependent metabolic switch, redirecting glucose flux from extracellular 2-keto-D-gluconic acid (2-KGA) biosynthesis to intracellular catabolism to support cellular survival. Key ED pathway enzymes (Zwf-1, KguD) were upregulated, while extracellular 2-KGA synthetic enzyme Gcd was downregulated. The remodeling of the respiratory chain, including cytochrome bc<sub>1</sub> complexes and terminal oxidases, enhanced electron transport efficiency as a coordinated adaptive response. These findings reveal the proteomic basis of thermal adaptation in <i>P. plecoglossicida</i> and provide molecular targets for engineering thermotolerant strains to optimize industrial 2-KGA fermentation.</p>

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Proteomics analysis of Pseudomonas plecoglossicida under thermal stress: the Entner-Doudoroff pathway as a temperature-dependent metabolic switch for thermal adaptation

  • 期刊论文

  • 发表日期：

    2026年7月4日

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  • Da-Ming Wang,
  • Yu-Ming Jing,
  • Qing-Hong Wang,
  • Lei Sun,
  • Wen-Jing Sun

摘要

Pseudomonas plecoglossicida is an industrial strain for the production of 2-keto-D-gluconic acid (2-KGA); however, its fermentation performance and 2-KGA yield significantly decline under high-temperature conditions. Tandem mass tag (TMT)-based quantitative proteomics was performed to investigate thermal adaptation mechanisms in Pseudomonas plecoglossicida at 32 °C, 36 °C, and 40 °C. In total, 565 and 1,665 differentially expressed proteins (DEPs) were identified at 36 °C and 40 °C vs. 32 °C, respectively. Functional enrichment revealed significant proteomic rewiring in carbohydrate metabolism, oxidative phosphorylation, and ubiquinone biosynthesis under thermal stress. The Entner-Doudoroff (ED) pathway acts as a temperature-dependent metabolic switch, redirecting glucose flux from extracellular 2-keto-D-gluconic acid (2-KGA) biosynthesis to intracellular catabolism to support cellular survival. Key ED pathway enzymes (Zwf-1, KguD) were upregulated, while extracellular 2-KGA synthetic enzyme Gcd was downregulated. The remodeling of the respiratory chain, including cytochrome bc1 complexes and terminal oxidases, enhanced electron transport efficiency as a coordinated adaptive response. These findings reveal the proteomic basis of thermal adaptation in P. plecoglossicida and provide molecular targets for engineering thermotolerant strains to optimize industrial 2-KGA fermentation.