# Evaluation of the Novel Bacteriophage Chage1 and Its Endolysin LysCG1 as Biocontrol Agents against Cronobacter sakazakii in Foods

## TL;DR

This study explores a new bacteriophage and its enzyme to control a harmful bacteria in infant formula and food products.

## Contribution

A novel bacteriophage (Chage1) and its endolysin (LysCG1) are evaluated for biocontrol of Cronobacter sakazakii in food.

## Key findings

- Chage1 is stable across a wide range of temperatures and pH levels and inhibits C. sakazakii growth for up to 24 hours.
- LysCG1, derived from Chage1, shows broader antibacterial activity against multiple Gram-negative bacteria.
- Chage1 and LysCG1 demonstrate antibacterial effects in powdered infant formula and milk samples.

## Abstract

Cronobacter sakazakii is a foodborne pathogen transmitted through powdered infant formula (PIF) and consequently leads to illness in infants. Owing to the pathogen’s occasional antibiotic resistance, bacteriophages offer a promising strategy for controlling this pathogen. Therefore, this study characterized a novel virulent phage (Chage1) that specifically infects C. sakazakii cells. Chage1 exhibited stability across a temperature range of 4–50°C and pH conditions of 4–11. Moreover, Chage1 inhibited bacterial growth for up to 24 h when inoculated into C. sakazakii at a multiplicity of infection (MOI) of 1. Chage1 (MOI of 105) also showed antibacterial activity against two C. sakazakii strains in PIF and milk samples for 8 h at 37°C. Whole genome sequencing of Chage1 revealed a 52,653 bp DNA genome with 72 predicted open reading frames (ORF) containing an endolysin gene (ORF 60). The endolysin LysCG1 was fused with a maltose-binding protein for solubility and its lytic activity against various bacterial strains was evaluated. LysCG1 exhibited a broader lysis spectrum than that of Chage1 by inhibiting the growth of diverse ethylenediaminetetraacetic acid-pretreated Gram-negative bacterial cells. Overall, this study provides a basis for the potential application of Chage1 and its endolysin for the control of C. sakazakii in food production.