Bifunctional fusion protein for biofilm degradation and quorum sensing disruption

Background

Bacterial biofilms pose a major challenge in clinical and industrial environments due to their dense extracellular matrix and heightened tolerance to antibiotics and immune responses. Infections involving biofilms, particularly those caused by Pseudomonas aeruginosa, are difficult to eradicate and contribute to chronic wounds, persistent medical device contamination, and severe respiratory conditions such as those seen in cystic fibrosis.

Because antibiotics struggle to penetrate the protective EPS matrix and many biofilm-embedded bacteria enter dormant states that reduce drug susceptibility, current treatments often fail to fully clear infections, leading to recurrence and escalating resistance. Physical removal is invasive and frequently impractical, and quorum sensing inhibitors alone often lack sufficient potency or broad applicability. These limitations underscore the need for a multifaceted therapeutic strategy capable of dismantling the biofilm matrix while simultaneously disrupting bacterial communication pathways essential for biofilm maintenance and regrowth.

Technology overview

This technology introduces a bifunctional fusion protein that integrates two synergistic enzymatic activities to eliminate Pseudomonas aeruginosa biofilms. The construct combines a truncated PslG amylase that degrades the polysaccharide-rich biofilm matrix with an engineered AiiA lactonase that hydrolyzes AHL quorum sensing molecules to prevent biofilm formation and reduce virulence.

Catalytic efficiency of the AiiA domain is significantly enhanced by replacing native zinc ions with cobalt, resulting in a 90- to 100-fold activity increase. A thioredoxin tag at the N-terminus improves solubility and stability, and a flexible linker and multiple affinity tags facilitate efficient purification. Produced in E. coli and refined through multi-step chroma­tography, the fusion protein demonstrates strong biofilm disruption and quorum quenching in functional assays.

By simultaneously degrading biofilm structure and inhibiting communication pathways critical for persistence, the technology offers a comprehensive, high-potency solution suitable for therapeutic or device-based applications.

Benefits

• Dual-action mechanism disrupts biofilm structure and bacterial signaling.
• Cobalt-enhanced AiiA lactonase provides a 90- to 100-fold catalytic efficiency increase.
• Thioredoxin tag improves solubility and stability for reliable expression.
• Effective against persistent Pseudomonas aeruginosa biofilms
• Enables potential synergy with existing antibiotics

Applications

• Chronic wound treatment
• Medical device coatings
• Respiratory infection therapeutics
• Industrial biofilm management
• Anti-biofilm research tools

Opportunity

• Addresses unmet need for solutions that both degrade biofilms and inhibit their reformation
• Provides a potent, modular platform adaptable to gram-negative pathogens and combination therapies
• Strong potential for integration into topical, inhaled, or device-based antimicrobial products
• Available for exclusive licensing

Intellectual property

U.S. Provisional application serial no. 63/688,574 filed on 08/29/2024

PCT serial no. PCT/US2025/043932 filed on 08/28/2025