# Heterologous Production of Antimicrobial Peptides in Yeast Allows for Massive Assessment of the Activity of DNA-Encoded Antimicrobials In Situ

## TL;DR

This paper describes a method using yeast to mass-produce and screen antimicrobial peptides, helping identify promising new antibiotics.

## Contribution

A novel platform for high-throughput screening of DNA-encoded antimicrobial peptides using engineered yeast.

## Key findings

- Yeast clones producing thanatin and protegrin-1 showed the highest antimicrobial activity.
- Recombinant thanatin production was higher and less toxic compared to protegrin-1.
- The method enables rapid identification of antimicrobial agents with improved therapeutic potential.

## Abstract

Antibiotic resistance threatens global healthcare. In clinical practice,
conventional antibiotics are becoming gradually less effective. Moreover, the
introduction of new antimicrobial agents into clinical practice leads to the
emergence of resistant pathogenic strains within just a few years. Hence, the
development of platforms for massive creation and screening of new
antimicrobial agents is of particular importance. Massive parallel screening
will greatly reduce the time required to identify the most promising drug
candidates. Meanwhile, DNA-encoded antimicrobial agents offer unique
opportunities for the high-throughput development of new antibiotics. Here, the
yeast Pichia pastoris was engineered to produce a panel of
antimicrobial peptides (AMPs), followed by high-throughput screening of AMP
producers that inhibit bacterial growth in situ. Yeast clones
producing thanatin and protegrin-1 exhibited the highest level of antimicrobial
activity among the panel of AMPs under investigation. The production level of
recombinant thanatin was significantly higher than that of protegrin-1, which
correlates with its low toxicity. The designed technique of massive assessment
of the activity of DNA-encoded antimicrobial agents enables the identification
of drug candidates with an increased therapeutic index. Further development of
methods for a rational design of artificial diversity in AMPs, followed by deep
functional profiling of antimicrobial activity, will yield new AMPs with
improved therapeutic characteristics.