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<p>The tightly regulated central carbon metabolism in <i>Saccharomyces cerevisiae</i>, intricately linked to carbon sources utilized, poses a significant challenge to engineering efforts aimed at increasing the flux through its different pathways. Here, we present a modular deregulation strategy that enables high conversion rates of xylose through the central carbon metabolism. Specifically, employing a multifaceted approach encompassing five different engineering strategies—promoter engineering, transcription factor manipulation, biosensor construction, introduction of heterologous enzymes, and expression of mutant enzymes we engineer different modules of the central carbon metabolism at both the genetic and enzymatic levels. This leads to an enhanced conversion rate of xylose into acetyl-CoA-derived products, with 3-hydroxypropionic acid (3–HP) serving as a representative case in this study. By implementing a combination of these approaches, the developed yeast strain demonstrates a remarkable enhancement in 3–HP productivity, achieving a 4.7–fold increase when compared to our initially optimized 3–HP producing strain grown on xylose as carbon source. These results illustrate that the rational engineering of yeast central metabolism is a viable approach for boosting the metabolic flux towards acetyl–CoA–derived products on a non-glucose carbon source.</p>

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Modular deregulation of central carbon metabolism for efficient xylose utilization in Saccharomyces cerevisiae

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2025年5月16日

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  • Xiaowei Li,
  • Yanyan Wang,
  • Xin Chen,
  • Leon Eisentraut,
  • Chunjun Zhan,
  • Jens Nielsen,
  • Yun Chen

摘要

The tightly regulated central carbon metabolism in Saccharomyces cerevisiae, intricately linked to carbon sources utilized, poses a significant challenge to engineering efforts aimed at increasing the flux through its different pathways. Here, we present a modular deregulation strategy that enables high conversion rates of xylose through the central carbon metabolism. Specifically, employing a multifaceted approach encompassing five different engineering strategies—promoter engineering, transcription factor manipulation, biosensor construction, introduction of heterologous enzymes, and expression of mutant enzymes we engineer different modules of the central carbon metabolism at both the genetic and enzymatic levels. This leads to an enhanced conversion rate of xylose into acetyl-CoA-derived products, with 3-hydroxypropionic acid (3–HP) serving as a representative case in this study. By implementing a combination of these approaches, the developed yeast strain demonstrates a remarkable enhancement in 3–HP productivity, achieving a 4.7–fold increase when compared to our initially optimized 3–HP producing strain grown on xylose as carbon source. These results illustrate that the rational engineering of yeast central metabolism is a viable approach for boosting the metabolic flux towards acetyl–CoA–derived products on a non-glucose carbon source.