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<p>The biosynthesis of surfactin, a potent lipopeptide biosurfactant produced by <i>Bacillus subtilis</i>, imposes a significant metabolic burden on the host organism. Understanding the metabolic costs associated with surfactin production, specifically the ATP demand and precursor diversion resulting from the expression of the large <i>srfAA-AD</i> operon (<i>srfA</i> operon), is crucial for optimizing production strains. In this study, the metabolic burden and growth impacts associated with surfactin biosynthesis in <i>B. subtilis</i> were quantitatively evaluated by comparing a reference surfactin-producing strain (BMV9) with two mutant strains: BMV12, which lacks the <i>srfA</i> operon, and BMV33, which retains the <i>srfA</i> operon but lacks the <i>sfp</i> gene. Our analysis included theoretical calculations of ATP and NADPH + H<sup>+</sup> requirements for <i>de novo</i> surfactin synthesis, and we measured growth behavior and carbon and nitrogen source consumption. Results indicated that surfactin production significantly reduces biomass yield (<i>Y</i><sub><i>X/S</i></sub>) and specific growth rates due to the metabolic costs of expressing the non-ribosomal peptide synthetase (NRPS) and the diversion of key precursors. Notably, BMV12 exhibited higher growth rates compared to the surfactin-producing strain. Proteome analyses further revealed differential protein abundance in non-surfactin-producing strains, indicating altered metabolic pathways that may relieve the metabolic burden associated with surfactin synthesis. These findings highlight the complex trade-offs between secondary metabolite production and cellular growth, providing a foundation for metabolic engineering strategies aimed at optimizing surfactin yields while minimizing metabolic costs.</p>

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Exploring the metabolic burden of surfactin biosynthesis and the metabolic costs of srfA operon expression in Bacillus subtilis

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2026年6月19日

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  • Elvio Henrique Benatto Perino,
  • Eric Hiller,
  • Maliheh Vahidinasab,
  • Sandra Moneta,
  • Bahar Abrishamchi,
  • Kowsala Nagendran,
  • Philipp Hubel,
  • Jens Pfannstiel,
  • Rudolf Hausmann

摘要

The biosynthesis of surfactin, a potent lipopeptide biosurfactant produced by Bacillus subtilis, imposes a significant metabolic burden on the host organism. Understanding the metabolic costs associated with surfactin production, specifically the ATP demand and precursor diversion resulting from the expression of the large srfAA-AD operon (srfA operon), is crucial for optimizing production strains. In this study, the metabolic burden and growth impacts associated with surfactin biosynthesis in B. subtilis were quantitatively evaluated by comparing a reference surfactin-producing strain (BMV9) with two mutant strains: BMV12, which lacks the srfA operon, and BMV33, which retains the srfA operon but lacks the sfp gene. Our analysis included theoretical calculations of ATP and NADPH + H+ requirements for de novo surfactin synthesis, and we measured growth behavior and carbon and nitrogen source consumption. Results indicated that surfactin production significantly reduces biomass yield (YX/S) and specific growth rates due to the metabolic costs of expressing the non-ribosomal peptide synthetase (NRPS) and the diversion of key precursors. Notably, BMV12 exhibited higher growth rates compared to the surfactin-producing strain. Proteome analyses further revealed differential protein abundance in non-surfactin-producing strains, indicating altered metabolic pathways that may relieve the metabolic burden associated with surfactin synthesis. These findings highlight the complex trade-offs between secondary metabolite production and cellular growth, providing a foundation for metabolic engineering strategies aimed at optimizing surfactin yields while minimizing metabolic costs.