Multiplexed bacteriocin synthesis to combat and prevent antimicrobial resistance

Area of Science:

• Biotechnology
• Microbiology
• Drug Discovery

Background:

• Bacteriocins offer a promising avenue to address antimicrobial resistance (AMR) due to their natural origin and targeted activity.
• Developing effective bacteriocin-based therapies is crucial for combating infections caused by multidrug-resistant pathogens.

Purpose of the Study:

• To rapidly produce bacteriocin cocktails using cell-free gene expression (CFE) for combating AMR.
• To design targeted bacteriocin combinations that prevent resistance development and enhance efficacy.
• To evaluate the efficacy of bacteriocin cocktails against multidrug-resistant pathogens both in vitro and in vivo.

Main Methods:

• Engineered DNA devices and CFE were employed for rapid synthesis of linear and circular bacteriocin cocktails.
• Bacteriocin pathways were leveraged for designing cocktails with enhanced cell envelope penetration.
• Continuous exchange CFE, DNA part reengineering, and disulfide bond formation optimization were utilized.
• Efficacy was tested against multidrug-resistant human pathogens and in the Galleria mellonella animal model.

Main Results:

• Tailored bacteriocin cocktails effectively eradicated target bacteria and prevented resistance development.
• Optimized synthesis using CFE and reengineered DNA parts improved bacteriocin production.
• Bacteriocin mixtures demonstrated efficacy against various multidrug-resistant human pathogens.
• In vivo testing in Galleria mellonella confirmed the potential of bacteriocin cocktails.

Conclusions:

• Bacteriocin cocktails produced via CFE represent a viable strategy for innovative treatments against multidrug-resistant infections.
• The developed platform facilitates rapid expression and testing of bacteriocin combinations for therapeutic applications.
• Bacteriocins hold significant potential as a next-generation therapeutic approach to combat the AMR crisis.