# A 2000-Year-Old Bacillus stercoris Strain Sheds Light on the Evolution of Cyclic Antimicrobial Lipopeptide Synthesis

## TL;DR

A 2000-year-old strain of Bacillus stercoris provides insights into the evolution of antimicrobial lipopeptide synthesis in bacteria.

## Contribution

The study reveals the genome of an ancient Bacillus stercoris strain and its antimicrobial gene conservation.

## Key findings

- The ancient Bacillus stercoris strain Mal05 has genes for fengycin and surfactin biosynthesis.
- These antimicrobial genes are present in all sequenced B. stercoris strains but not under strong purifying selection.
- The genes may provide a competitive advantage without being essential for bacterial fitness.

## Abstract

Some bacteria (notably the genera Bacillus and Clostridium) have the capacity to form endospores that can survive for millions of years in isolated habitats. The genomes of such ancient bacteria provide unique opportunities to understand bacterial evolution and metabolic capabilities over longer time scales. Herein, we sequenced the genome of a 2000-year-old bacterial strain (Mal05) isolated from intact apple seeds recovered during archaeological excavations of a Roman villa in Italy. Phylogenomic analyses revealed that this strain belongs to the species Bacillus stercoris and that it is placed in an early-branching position compared to most other strains of this species. Similar to other Bacillus species, B. stercoris Mal05 had been previously shown to possess antifungal activity. Its genome encodes all the genes necessary for the biosynthesis of fengycin and surfactin, two cyclic lipopeptides known to play a role in the competition of Bacilli with other microorganisms due to their antimicrobial activity. Comparative genomics and analyses of selective pressure demonstrate that these genes are present in all sequenced B. stercoris strains, despite the fact that they are not under strong purifying selection. Hence, these genes may not be essential for the fitness of these bacteria, but they can still provide a competitive advantage against other microorganisms present in the same environment.