High-Resolution Melting Analysis Potential for <i>Saccharomyces cerevisiae</i> var. <i>boulardii</i> Authentication in Probiotic-Enriched Food Matrices
To date, the only probiotic yeast with evidence of health-promoting effects is <i>Saccharomyces cerevisiae</i> var. <i>boulardii</i>. The expanded market including dietary supplements and functional foods supplemented with <i>Saccharomyces cerevisiae</i> var. <...
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| Main Authors: | , , |
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| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2024-11-01
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| Series: | BioTech |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2673-6284/13/4/48 |
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| Summary: | To date, the only probiotic yeast with evidence of health-promoting effects is <i>Saccharomyces cerevisiae</i> var. <i>boulardii</i>. The expanded market including dietary supplements and functional foods supplemented with <i>Saccharomyces cerevisiae</i> var. <i>boulardii</i> creates an environment conductive to food adulterations, necessitating rapid testing to verify product probiotic status. Herein, qPCR-HRM analysis was tested for probiotic yeast identification. The effectiveness of the primer pairs’ set was examined, designed to amplify heterogeneous regions in (a) rDNA sequences previously designed to identify food-derived yeast and (b) genes associated with physiological and genotypic divergence of <i>Saccharomyces cerevisiae</i> var. <i>boulardii.</i> Preliminary tests of amplicons’ differentiation power enabled the selection of interspecies sequences for <i>18SrRNA</i> and ITS and genus-specific sequences <i>HO</i>, <i>RPB2</i>, <i>HXT9</i> and <i>MAL11.</i> The multi-fragment qPCR-HRM analysis was sufficient for culture-dependent <i>Saccharomyces cerevisiae</i> var. <i>boulardii</i> identification and proved effective in the authentication of dietary supplements’ probiotic composition. The identification of <i>S. cerevisiae</i> var. <i>boulardii</i> in complex microbial mixtures of kefir succeeded with more specific intragenus sequences <i>HO</i> and <i>RPB2.</i> The predominance of <i>S. cerevisiae</i> var. <i>boulardii</i> in the tested matrices, quantitatively corresponded to the probiotic-enriched food, was crucial for identification with qPCR–HRM analysis. Considering the reported assumptions, qPCR-HRM analysis is an appropriate tool for verifying probiotic-enriched food. |
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| ISSN: | 2673-6284 |