Proteomic Analysis of Listeria monocytogenes FBUNT During Biofilm Formation at 10°C in Response to Lactocin AL705.

Constanza Melian,Lucía M Mendoza,Franco Segli,Graciela Margarita Vignolo,Patricia Castellano

doi:10.3389/fmicb.2021.604126

Constanza Melian, Lucía M Mendoza + Show 3 more

Open Access

https://doi.org/10.3389/fmicb.2021.604126

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Abstract

Listeria monocytogenes is one of the major food-related pathogens and is able to survive and multiply under different stress conditions. Its persistence in industrial premises and foods is partially due to its ability to form biofilm. Thus, as a natural strategy to overcome L. monocytogenes biofilm formation, the treatment with lactocin AL705 using a sublethal dose (20AU/ml) was explored. The effect of the presence of the bacteriocin on the biofilm formation at 10°C of L. monocytogenes FBUNT was evaluated for its proteome and compared to the proteomes of planktonic and sessile cells grown at 10°C in the absence of lactocin. Compared to planktonic cells, adaptation of sessile cells during cold stress involved protein abundance shifts associated with ribosomes function and biogenesis, cell membrane functionality, carbohydrate and amino acid metabolism, and transport. When sessile cells were treated with lactocin AL705, proteins’ up-regulation were mostly related to carbohydrate metabolism and nutrient transport in an attempt to compensate for impaired energy generation caused by bacteriocin interacting with the cytoplasmic membrane. Notably, transport systems such as β-glucosidase IIABC (lmo0027), cellobiose (lmo2763), and trehalose (lmo1255) specific PTS proteins were highly overexpressed. In addition, mannose (lmo0098), a specific PTS protein indicating the adaptive response of sessile cells to the bacteriocin, was downregulated as this PTS system acts as a class IIa bacteriocin receptor. A sublethal dose of lactocin AL705 was able to reduce the biofilm formation in L. monocytogenes FBUNT and this bacteriocin induced adaptation mechanisms in treated sessile cells. These results constitute valuable data related to specific proteins targeting the control of L. monocytogenes biofilm upon bacteriocin treatment.

Highlights

Listeria monocytogenes is the causative agent of the severe human and animal disease listeriosis, which has a high mortality rate (Listeriosis, 2017; Foodnet Centers for Disease Control and Prevention, 2018) and whose features include gastroenteritis in healthy individuals that can develop into a severe invasive illness in the elderly, pregnant women, infants, and immunocompromised (Ferreira et al, 2014)
The analysis of L. monocytogenes FBUNT proteome was performed in order to elucidate the regulation of proteins during biofilm formation at 10°C as well as the effect of lactocin AL705
In accordance with a previous study, the calculated MIC of lactocin AL705 against L. monocytogenes FBUNT was 40 arbitrary units per milliliter (AU/ml) (7.4 μg/ml), whereas the subinhibitory concentration of 20 AU/ml caused the highest inhibition of biofilm (75%) but had no effect on the growth of the planktonic population (Melian et al, 2019)

Summary

Introduction

Listeria monocytogenes is the causative agent of the severe human and animal disease listeriosis, which has a high mortality rate (Listeriosis, 2017; Foodnet Centers for Disease Control and Prevention, 2018) and whose features include gastroenteritis in healthy individuals that can develop into a severe invasive illness in the elderly, pregnant women, infants, and immunocompromised (Ferreira et al, 2014) This foodborne pathogen is implicated in sporadic cases, outbreaks, and food recalls worldwide (Desai et al, 2019). Ready-to-eat (RTE) foods are common vehicles involved in listeriosis outbreaks, this stimulating many countries to create specific legislation aimed at controlling L. monocytogenes in RTE foods after the 1990s (Camargo et al, 2017) The persistence of this pathogen in food environments is partly due to its ability to form biofilms; bacterial adhesion to industrial surfaces is of great concern as a chronic source of contamination (Bridier et al, 2015; Coughlan et al, 2016). There is abundant evidence indicating that pathogens capable of forming biofilm are self-protected from common cleaning procedures, allowing them to remain in the environment postsanitation and increasing their resistance to antimicrobials/ sanitizers (Bae et al, 2012; Rodríguez-López et al, 2018)

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