Genetic Diversity of Leuconostoc mesenteroides Isolates from Traditional Montenegrin Brine Cheese.

Werner Ruppitsch,Jasmin Sucher,Anna Stöger,Aleksandra Martinović,Andjela Nisic,Adriana Cabal,Franz Allerberger,Patrick Hyden

doi:10.3390/microorganisms9081612

Abstract

In many dairy products, Leuconostoc spp. is a natural part of non-starter lactic acid bacteria (NSLAB) accounting for flavor development. However, data on the genomic diversity of Leuconostoc spp. isolates obtained from cheese are still scarce. The focus of this study was the genomic characterization of Leuconostoc spp. obtained from different traditional Montenegrin brine cheeses with the aim to explore their diversity and provide genetic information as a basis for the selection of strains for future cheese production. In 2019, sixteen Leuconostoc spp. isolates were obtained from white brine cheeses from nine different producers located in three municipalities in the northern region of Montenegro. All isolates were identified as Ln. mesenteroides. Classical multilocus sequence tying (MLST) and core genome (cg) MLST revealed a high diversity of the Montenegrin Ln. mesenteroides cheese isolates. All isolates carried genes of the bacteriocin biosynthetic gene clusters, eight out of 16 strains carried the citCDEFG operon, 14 carried butA, and all 16 isolates carried alsS and ilv, genes involved in forming important aromas and flavor compounds. Safety evaluation indicated that isolates carried no pathogenic factors and no virulence factors. In conclusion, Ln. mesenteroides isolates from Montenegrin traditional cheeses displayed a high genetic diversity and were unrelated to strains deposited in GenBank.

Highlights

Introduction published maps and institutional affilThe genus Leuconostoc (Ln.) currently comprises 14 species and eight subspecies, all being Gram-positive, non-spore-forming, non-motile Lactic Acid Bacteria (LAB)
The aim of this study was to explore the genetic diversity of Ln. mesenteroides isolates obtained from Montenegrin brine cheeses using whole genome sequencing to prove the origin of the Montenegrin strains that can be used in future industrial cheese production
Inhz 21 mm Traditional food products are an acknowledged part of the identity, culture, and heritage of a country and, in addition, they may serve as a valuable resource for future food production [16,17,52]

Summary

Introduction

The genus Leuconostoc (Ln.) currently comprises 14 species and eight subspecies, all being Gram-positive, non-spore-forming, non-motile Lactic Acid Bacteria (LAB). Within the genus Leuconostoc, Ln. mesenteroides, Ln. pseudomesenteroides and Ln. lactis have their role in food fermentation and can be isolated from various food-related ecological niches, including beverages, meat and dairy products, and some plant materials, implying wide distribution and specialized adaptation to these diverse environments [1,2,3,4]. Ln. mesenteroides due to the common properties they shared [6] and their high degree of relatedness shown by DNA—DNA hybridization [5]. Leuconostoc spp. are usually applied as an adjunct culture in combination with the fast acid producing Lactococcus spp., as undefined mixed (DL) iations

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