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<p>Bacterial biofilms formed by phytopathogens confer formidable resistance to chemical pesticides, underscoring an urgent need for innovative antimicrobial solutions. Antimicrobial peptides (AMPs), exemplified by the potent bee venom derivative melittin, offer a promising alternative owing to their broad-spectrum activity and intrinsic biofilm-disrupting capacity. However, the agricultural application of melittin is severely hindered by rapid environmental degradation, susceptibility to enzymatic degradation, and non-selective cytotoxicity. Here, we report a metal-coordination-driven nanoassembly strategy to enhance the stability and efficacy of melittin. Engineering an N-terminal hexahistidine tag enabled a one-step assembly of melittin into uniform nanoparticles (NanoMel) via Zn²⁺ coordination. This nanoformulation improved the antibacterial potency, lowering the half-maximal effective concentration (EC₅₀) values against <i>Xanthomonas oryzae pv. oryzae</i> (<i>Xoo</i>), <i>Xanthomonas oryzae pv. oryzicola</i> (<i>Xoc</i>) to 3.795&#xa0;µg/mL and 3.202&#xa0;µg/mL, representing a 1.59- and 1.38-fold enhancement over its linear counterpart. Furthermore, NanoMel demonstrated superior biofilm eradication, degrading 86.9% of mature <i>Xoo</i> biofilms at 24&#xa0;µg/mL, significantly outperforming the free peptide. <i>In planta</i> assays revealed that NanoMel provided 68.2% curative and 65.9% protective efficacy against rice bacterial leaf blight at 200&#xa0;µg/mL, surpassing the commercial bactericide thiodiazole-copper 20% suspension concentrate (TC-20% SC). Furthermore, the nanoassemblies effectively attenuated the inherent toxicity of melittin, as evidenced by significantly improved safety profiles in the zebrafish model. Collectively, these findings establish the metal-coordination-driven nanoassembly as a platform for constructing effective and eco-friendly AMP-based bionanobactericides, demonstrating a potent and practical strategy for sustainable plant protection.</p> Graphical Abstract <p></p>

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Metal-driven nanoassembly of hexahistidine-tagged melittin enables superior phytopathogen biofilm degradation with attenuated toxicity

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2026年6月23日

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  • Xingcheng Zhou,
  • Jie Hong,
  • Li Yang,
  • Satyabrata Nanda,
  • Youbo Yu,
  • You Wang,
  • Krongthong Kamonsuangkasem,
  • Ming Chang,
  • Janaki Mohotti,
  • Gefei Hao,
  • Peiyi Wang,
  • Libo Zhang

摘要

Bacterial biofilms formed by phytopathogens confer formidable resistance to chemical pesticides, underscoring an urgent need for innovative antimicrobial solutions. Antimicrobial peptides (AMPs), exemplified by the potent bee venom derivative melittin, offer a promising alternative owing to their broad-spectrum activity and intrinsic biofilm-disrupting capacity. However, the agricultural application of melittin is severely hindered by rapid environmental degradation, susceptibility to enzymatic degradation, and non-selective cytotoxicity. Here, we report a metal-coordination-driven nanoassembly strategy to enhance the stability and efficacy of melittin. Engineering an N-terminal hexahistidine tag enabled a one-step assembly of melittin into uniform nanoparticles (NanoMel) via Zn²⁺ coordination. This nanoformulation improved the antibacterial potency, lowering the half-maximal effective concentration (EC₅₀) values against Xanthomonas oryzae pv. oryzae (Xoo), Xanthomonas oryzae pv. oryzicola (Xoc) to 3.795 µg/mL and 3.202 µg/mL, representing a 1.59- and 1.38-fold enhancement over its linear counterpart. Furthermore, NanoMel demonstrated superior biofilm eradication, degrading 86.9% of mature Xoo biofilms at 24 µg/mL, significantly outperforming the free peptide. In planta assays revealed that NanoMel provided 68.2% curative and 65.9% protective efficacy against rice bacterial leaf blight at 200 µg/mL, surpassing the commercial bactericide thiodiazole-copper 20% suspension concentrate (TC-20% SC). Furthermore, the nanoassemblies effectively attenuated the inherent toxicity of melittin, as evidenced by significantly improved safety profiles in the zebrafish model. Collectively, these findings establish the metal-coordination-driven nanoassembly as a platform for constructing effective and eco-friendly AMP-based bionanobactericides, demonstrating a potent and practical strategy for sustainable plant protection.

Graphical Abstract