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<p>The concept of a methanol-based circular bioeconomy envisions the conversion of greenhouse gases into one-carbon compounds, which are subsequently utilized to produce high-value-added products. However, the toxicity of intermediates limits the metabolic efficiency within chassis strains. Here a carbon metabolism reconfiguration strategy combined with adaptive laboratory evolution was applied to engineer an artificial methylotrophic yeast by accelerating the flux of toxic intermediates into the central carbon metabolism. After modifying the key gene <i>Sfa1</i> identified by adaptive laboratory evolution and introducing a light-responsive switch to dynamically redirect the carbon flux, the engineered strain not only restored its growth in minimal medium with 2% methanol, but also synthesized chondroitin sulfate A driven by the dark reaction. Furthermore, we designed a 3′-phosphoadenosine-5′-phosphosulfate supply strategy and energy adapter to increase the sulfonation level. This study illustrates the exciting potential of methylotrophic <i>Saccharomyces</i> <i>cerevisiae</i> as a platform for biosynthesis.</p><p></p>

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Constructing artificial methylotrophic S. cerevisiae for chondroitin sulfate production using a multidimensional combinatorial strategy

  • 期刊论文

  • 发表日期：

    2026年5月22日

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  • Lei Li,
  • Yixun Jiang,
  • Wenjie Ma,
  • Chenying Li,
  • Ruirui Shi,
  • Guangyao Tang,
  • Yaxin He,
  • Jiaqi Li,
  • Zepin Guo,
  • Yuxin Zhang,
  • Shuhong Mao,
  • Mingdong Yao,
  • Zhengqiang Jiang,
  • Fuping Lu,
  • Hui-Min Qin

摘要

The concept of a methanol-based circular bioeconomy envisions the conversion of greenhouse gases into one-carbon compounds, which are subsequently utilized to produce high-value-added products. However, the toxicity of intermediates limits the metabolic efficiency within chassis strains. Here a carbon metabolism reconfiguration strategy combined with adaptive laboratory evolution was applied to engineer an artificial methylotrophic yeast by accelerating the flux of toxic intermediates into the central carbon metabolism. After modifying the key gene Sfa1 identified by adaptive laboratory evolution and introducing a light-responsive switch to dynamically redirect the carbon flux, the engineered strain not only restored its growth in minimal medium with 2% methanol, but also synthesized chondroitin sulfate A driven by the dark reaction. Furthermore, we designed a 3′-phosphoadenosine-5′-phosphosulfate supply strategy and energy adapter to increase the sulfonation level. This study illustrates the exciting potential of methylotrophic Saccharomyces cerevisiae as a platform for biosynthesis.