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<p>RNA helicases are a class of highly conserved RNA-binding proteins that unwind double-stranded RNA by hydrolyzing ATP, playing essential roles in processes such as ribosome biogenesis, pre-mRNA splicing, transport, translation, and decay of pre-mRNA. However, their functional roles in the stress responses of <i>Candida glycerinogenes</i> remain uncharacterized to date. Herein, <i>CgDBP7</i> (<i>C. glycerinogenes</i> DEAD-box RNA helicase <i>DBP7</i>) and its downstream responsive gene <i>CgMAK5</i> (<i>C. glycerinogenes</i> ATP-dependent RNA helicase <i>MAK5</i>) were first characterized as dual negative regulators of high-salinity tolerance in <i>C. glycerinogenes</i>. Mechanistically, the enhanced high-salinity tolerance of the engineered strain <i>C. glycerinogenes-antiCgDBP7a-antiCgMAK5</i> was associated with intracellular reactive oxygen species (ROS) detoxification and glycerol biosynthesis. Glycerol acts as an osmoprotectant to balance osmotic pressure under high salt, while efficient ROS detoxification mitigates cellular oxidative damage, collectively boosting tolerance and metabolite production. Under high-salinity conditions, the stress-resistant engineered strain <i>C. glycerinogenes-antiCgDBP7a-antiCgMAK5</i> achieved a 13.5% and 16.5% increase in glycerol and ethanol titers, respectively. In undetoxified lignocellulosic hydrolysate, this recombinant strain exhibited a further increase of 45.2% in ethanol titer and 13.4% in glycerol titer. Collectively, these findings demonstrate that <i>CgDBP7</i> and <i>CgMAK5</i> form a two-layered repressive circuit and serve as key genetic elements for salinity stress adaptation in <i>C. glycerinogenes,</i> providing programmable targets for engineering robust industrial yeast.</p>

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RNA helicase gene CgDBP7 improves the high salt tolerance of Candida glycerinogenes by regulating CgMAK5

  • 期刊论文

  • 发表日期：

    2026年4月13日

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  • Chungui Li,
  • Xiaoqing Hao,
  • Bin zhuge,
  • Hong Zong,
  • Ge Huang,
  • Yifan Wu,
  • Jiajun Peng,
  • Tong Li,
  • Kaina Wei,
  • Xinyao Lu

摘要

RNA helicases are a class of highly conserved RNA-binding proteins that unwind double-stranded RNA by hydrolyzing ATP, playing essential roles in processes such as ribosome biogenesis, pre-mRNA splicing, transport, translation, and decay of pre-mRNA. However, their functional roles in the stress responses of Candida glycerinogenes remain uncharacterized to date. Herein, CgDBP7 (C. glycerinogenes DEAD-box RNA helicase DBP7) and its downstream responsive gene CgMAK5 (C. glycerinogenes ATP-dependent RNA helicase MAK5) were first characterized as dual negative regulators of high-salinity tolerance in C. glycerinogenes. Mechanistically, the enhanced high-salinity tolerance of the engineered strain C. glycerinogenes-antiCgDBP7a-antiCgMAK5 was associated with intracellular reactive oxygen species (ROS) detoxification and glycerol biosynthesis. Glycerol acts as an osmoprotectant to balance osmotic pressure under high salt, while efficient ROS detoxification mitigates cellular oxidative damage, collectively boosting tolerance and metabolite production. Under high-salinity conditions, the stress-resistant engineered strain C. glycerinogenes-antiCgDBP7a-antiCgMAK5 achieved a 13.5% and 16.5% increase in glycerol and ethanol titers, respectively. In undetoxified lignocellulosic hydrolysate, this recombinant strain exhibited a further increase of 45.2% in ethanol titer and 13.4% in glycerol titer. Collectively, these findings demonstrate that CgDBP7 and CgMAK5 form a two-layered repressive circuit and serve as key genetic elements for salinity stress adaptation in C. glycerinogenes, providing programmable targets for engineering robust industrial yeast.