Continuous bioactivity-dependent evolution of an antibiotic biosynthetic pathway

Area of Science:

• Microbiology
• Synthetic Biology
• Biotechnology

Background:

• Antibiotic biosynthetic gene clusters (BGCs) are crucial for producing bioactive metabolites that confer a survival advantage.
• Natural selection drives the evolution of BGCs, adapting them for expression in diverse organisms.
• Understanding these evolutionary mechanisms can guide strategies for enhancing heterologous antibiotic production.

Purpose of the Study:

• To engineer antibiotic biosynthetic gene clusters (BGCs) for improved production in heterologous hosts.
• To demonstrate the utility of phage-assisted continuous evolution (PACE) for bioactivity-dependent BGC adaptation.
• To explore natural selection principles for optimizing metabolic pathways and product yields.

Main Methods:

• Phage-assisted continuous evolution (PACE) was employed to adapt the bicyclomycin (BCM) BGC.
• Bioactivity-dependent selection was used to drive the evolution of the BCM BGC.
• The adapted BGC was evaluated for improved production in a heterologous host.

Main Results:

• Successful adaptation of the BCM BGC for enhanced production in a heterologous host.
• Demonstrated the efficacy of PACE in achieving bioactivity-dependent evolution of BGCs.
• Achieved improved yields of bicyclomycin through engineered evolutionary pathways.

Conclusions:

• Engineered bioactivity-dependent evolution offers a powerful strategy for optimizing metabolic pathways.
• PACE can be utilized to rapidly adapt BGCs for improved heterologous expression.
• This approach provides novel routes for enhancing natural product yields and developing new antibiotics.