Multiplex-PCR Detection of an Atypical <i>Leuconostoc mesenteroides</i> subsp. <i>jonggajibkimchii</i> Phenotype Dominating the Terminal Spoilage Microbial Association of a Fresh Greek Whey Cheese Stored at 4 °C in Vacuum

Multiplex-PCR Detection of an Atypical <i>Leuconostoc mesenteroides</i> subsp. <i>jonggajibkimchii</i> Phenotype Dominating the Terminal Spoilage Microbial Association of a Fresh Greek Whey Cheese Stored at 4 °C in Vacuum

A species-specific multiplex-PCR method and phenotypic tests were combined to evaluate biochemical and genotypic differences between 24 representative <i>Leuconostoc mesenteroides</i> diverse isolates previously found to dominate in spoiled, vacuum-packed Anthotyros whey cheeses stored a...

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Main Authors: Nikoletta Sameli, Eleni Sioziou, Loulouda Bosnea, Spiros Paramithiotis, John Samelis
Format: Article
Language:English
Published: MDPI AG 2024-07-01
Series:Applied Microbiology
Subjects:
<i>Leuconostoc mesenteroides</i> subsp. <i>jonggajibkimchii</i>
subspecies identification
<i>rpoB</i>
multiplex-PCR
<i>araA</i>/<i>dsr</i>/<i>sorA</i> gene profiles
whey cheese spoilage
Online Access:https://www.mdpi.com/2673-8007/4/3/76
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Summary:A species-specific multiplex-PCR method and phenotypic tests were combined to evaluate biochemical and genotypic differences between 24 representative <i>Leuconostoc mesenteroides</i> diverse isolates previously found to dominate in spoiled, vacuum-packed Anthotyros whey cheeses stored at 4 °C for 40 days and identified by 16S rRNA gene sequencing. Based on their phenotypic (API 50 CHL) profiles, the 24 isolates comprised 6 multi-strain and 7 single-strain biotypes. Only two single-strain biotypes (L4A and L4B) produced slime (dextran) from sucrose, and only four biotypes (L2A–L2C, L3; 7 isolates) fermented L-arabinose; the remaining 15 isolates (biotypes L1A–L1F) were dextran-negative, oligofermenting <i>Ln. mesenteroides</i> variants, able to ferment D-xylose and grow at 37 °C. Based on their multiplex-PCR (<i>rpoB</i>, <i>araA</i>, <i>dsr</i>, and <i>sorA</i>) gene profiles in comparison with those of the type strains of the four <i>Ln. mesenteroides</i> subsp. <i>cremoris</i> (<i>rpoB</i>), <i>dextranicum</i> (<i>rpoB</i>/<i>dsr</i>), <i>mesenteroides</i> (<i>rpoB</i>/<i>araA</i>/<i>dsr</i>/<i>sorA</i>), and <i>jonggajibkimchii</i> (<i>rpoB</i>/<i>araA</i>/<i>dsr</i>), no isolate was assigned to the first two subspecies and only four isolates (L2A and L2C) to the subsp. <i>mesenteroides</i>. Ten isolates shared the subsp. <i>jonggajibkimchii</i> profile, while the other ten ones have a fifth atypical profile (<i>rpoB</i>/<i>dsr</i>/<i>sorA</i>), seemingly being closer to the subsp. <i>dextranicum</i>. Particularly the atypical biotype L1B representatives of the most prevalent psychrotrophic <i>Ln. mesenteroides</i> subsp. <i>jonggajibkimchii</i> (<i>rpoB</i>/<i>araA</i>/<i>dsr</i>) genotype at Anthotyros whey cheese spoilage deserve further biochemical and molecular characterization studies.
ISSN:2673-8007

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