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<p>Heterologous protein expression in <i>E. coli</i> facilitates high yield and quality, but environmental stress often destabilizes these proteins, affecting their activity. This study investigates a peptide co-expression system to enhance enzyme thermostability, demonstrated here using lipase as a model. Specifically, Late Abundant Embryogenesis (LEA) proteins were employed to protect proteins from damage. Recombinant lipase (MLipA) from <i>Bacillus licheniformis</i> was expressed alongside a LEA-like peptide, inspired by the 11 repetitive amino acid sequences of LEA proteins. Four LEA-like peptide co-expression systems were evaluated for their impact on lipase thermostability using a standard lipase assay. The purified recombinant MLipA was tested at 45&#xa0;°C, above its optimal temperature, revealing a two-fold increase in activity when co-expressed with the LEA-II-like peptide. Sequence analysis indicated that LEA-II contains more polar and aliphatic residues, enhancing the stability of the MLipA-LEA II complex at elevated temperatures. Molecular docking suggested that specific residues of LEA-II interact with lipase to facilitate enzyme–substrate complex formation with <i>p</i>NPL. Molecular dynamics simulations further confirmed that the MLipA-LEA II complex is more stable at 318.15&#xa0;K compared to MLipA alone. MM-PBSA analysis showed improved binding affinity of <i>p</i>NPL in the MLipA-LEA II complex. Together, these findings establish LEA-like peptide co-expression as a novel, sequence-independent strategy for enhancing enzyme thermostability, bridging experimental and computational approaches with potential applications in both industrial biocatalysis and therapeutic protein stabilization.</p> Graphical abstract <p></p>

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Late embryogenesis abundant (LEA) peptide co-expression enhances the thermostability of Bacillus licheniformis lipase: an experimental and computational study

  • 期刊论文

  • 发表日期：

    2026年7月22日

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  • Ammar Khazaal Kadhim Almansoori,
  • Kang Siang Yu,
  • Faisal Mohamad,
  • Shinya Ikeno,
  • Rajesh Bhimgonda Patil,
  • Georcki Ropón-Palacios,
  • Rashidah Abdul Rahim

摘要

Heterologous protein expression in E. coli facilitates high yield and quality, but environmental stress often destabilizes these proteins, affecting their activity. This study investigates a peptide co-expression system to enhance enzyme thermostability, demonstrated here using lipase as a model. Specifically, Late Abundant Embryogenesis (LEA) proteins were employed to protect proteins from damage. Recombinant lipase (MLipA) from Bacillus licheniformis was expressed alongside a LEA-like peptide, inspired by the 11 repetitive amino acid sequences of LEA proteins. Four LEA-like peptide co-expression systems were evaluated for their impact on lipase thermostability using a standard lipase assay. The purified recombinant MLipA was tested at 45 °C, above its optimal temperature, revealing a two-fold increase in activity when co-expressed with the LEA-II-like peptide. Sequence analysis indicated that LEA-II contains more polar and aliphatic residues, enhancing the stability of the MLipA-LEA II complex at elevated temperatures. Molecular docking suggested that specific residues of LEA-II interact with lipase to facilitate enzyme–substrate complex formation with pNPL. Molecular dynamics simulations further confirmed that the MLipA-LEA II complex is more stable at 318.15 K compared to MLipA alone. MM-PBSA analysis showed improved binding affinity of pNPL in the MLipA-LEA II complex. Together, these findings establish LEA-like peptide co-expression as a novel, sequence-independent strategy for enhancing enzyme thermostability, bridging experimental and computational approaches with potential applications in both industrial biocatalysis and therapeutic protein stabilization.

Graphical abstract