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<p><i>Spirulina</i>, a commercially significant cyanobacterium, also named as <i>Arthrospira</i> or <i>Limnospira</i>, is widely used in food, feed, and pharmaceutical industries due to its rich protein content, phycocyanin, and exopolysaccharides (EPS). The role of the <i>pilA</i> gene was studied to encode the major pilin subunit of the type IV pilus (T4P) system in EPS synthesis and cell aggregation using the industrial strain <i>Spirulina platensis</i> FACHB-439. Through targeted knockout of <i>pilA</i>, we demonstrated that mutants exhibited enhanced EPS production and improved auto-aggregation under static conditions, despite losing phototaxis properties. Transcriptomic analysis revealed the coordinated up-regulation of genes involved in synthesis of partial T4P components (PilB, HpsC, etc.) and EPS (e.g., glycosyltransferase), and nitrogen recycling (Urease E/F), indicating adaptive metabolic responses to T4P deficiency. These findings showed the presence of integrating regulators linking the T4P system with EPS biosynthesis, nitrogen metabolism, and redox homeostasis. This study demonstrated that genetic knockout of <i>pilA</i> can be used to enhance EPS production and aggregation, offering a proof-of-concept for trait improvement in <i>S. platensis</i>, and providing a theoretical foundation and technical roadmap for the future development of industrial algal strains with superior performance.</p>

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Knockout of pilA gene in Spirulina platensis enhances the exopolysaccharide production and auto-aggregation

  • 期刊论文

  • 发表日期：

    2026年2月3日

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  • Yuxi Han,
  • Xiujun Xie,
  • Songcui Wu,
  • Shurui Wang,
  • Shan Gao,
  • Wenhui Gu,
  • Guangce Wang

摘要

Spirulina, a commercially significant cyanobacterium, also named as Arthrospira or Limnospira, is widely used in food, feed, and pharmaceutical industries due to its rich protein content, phycocyanin, and exopolysaccharides (EPS). The role of the pilA gene was studied to encode the major pilin subunit of the type IV pilus (T4P) system in EPS synthesis and cell aggregation using the industrial strain Spirulina platensis FACHB-439. Through targeted knockout of pilA, we demonstrated that mutants exhibited enhanced EPS production and improved auto-aggregation under static conditions, despite losing phototaxis properties. Transcriptomic analysis revealed the coordinated up-regulation of genes involved in synthesis of partial T4P components (PilB, HpsC, etc.) and EPS (e.g., glycosyltransferase), and nitrogen recycling (Urease E/F), indicating adaptive metabolic responses to T4P deficiency. These findings showed the presence of integrating regulators linking the T4P system with EPS biosynthesis, nitrogen metabolism, and redox homeostasis. This study demonstrated that genetic knockout of pilA can be used to enhance EPS production and aggregation, offering a proof-of-concept for trait improvement in S. platensis, and providing a theoretical foundation and technical roadmap for the future development of industrial algal strains with superior performance.