Molecular and Physiological Diversity of Indigenous Yeasts Isolated from Spontaneously Fermented Wine Wort from Ilfov County, Romania

Abstract

(1) Background: Wine yeast research offers the possibility of isolating new strains with distinct metabolic properties due to the geographical location of the vineyard and the processes used in winemaking. Our study deals with the isolation and identification of six yeasts from spontaneously fermented wine wort from Romania and their characterization as new potential starter culture for traditional beverages, for food industry or biomedicine. (2) Materials and methods: The isolates were identified using conventional taxonomy tests, phenotypic phylogeny analysis (Biolog YT), MALDI-TOF mass spectrometry, PCR-RFLP, and sequencing of the ITS1-5,8S-ITS2 rDNA region. The capacity of the yeasts to grow under thermal, ionic, and osmotic stress was determined. The safe status was confirmed by testing virulence and pathogenicity factors. Assays were performed in order to evaluate the growth inhibition of Candida strains and determine the antimicrobial mechanism of action. (3) Results and discussions: The yeast isolates were identified as belonging to the Metschinikowia, Hanseniaspora, Torulaspora, Pichia, and Saccharomyces genera. All the isolates were able to develop under the tested stress conditions and were confirmed as safe. With the exception of S. cerevisiae CMGB-MS1-1, all the isolates showed good antimicrobial activity based on competition for iron ions or production of killer toxins. (4) Conclusions: The results revealed the resistance of our yeasts to environmental conditions related to industrial and biomedical applications and their high potential as starter cultures and biocontrol agents, respectively.

Probiotic Yeasts and How to Find Them—Polish Wines of Spontaneous Fermentation as Source for Potentially Probiotic Yeasts

“…As a safety aspect, no hemolytic activity was present in our wine-derived strains. That is a result comparable to those obtained by Corbu et al, 2023, and Fernández-Pacheco et al, 2021 [ 59 , 60 ]. However, the strains 27_Sacch_cerevisiae, 32_Pich_kluyveri and 34_Metsch_pulcherrima showed the ability to produce biogenic amines (BAs) that excludes them as candidates for potentially probiotic strains [ 43 ], despite their promising properties in previous assays, due to potential risks for consumers [ 61 , 62 ].…”

Achievements of Autochthonous Wine Yeast Isolation and Selection in Romania—A Review

“…Systematic studies were started only in later 1970s by Kontek et al (1975Kontek et al ( -1977. Later on, a few studies took into account the study of the vineyard or fermented grape must yeast biodiversity as a whole [46,62].…”

“…Some teams made use of MALDI-TOF mass spectrometry, especially that of Bărbulescu et al, wherein the isolated strains were prepared for the analysis after the extraction of peptides with formic acid, ethanol, and acetonitrile [49]. A similar approach was taken by Corbu and Csutak when studying yeast biodiversity in different traditional fermented foods, including wine [62]. For a more accurate physiological identification of the tested strains, phenotypic phylogeny analyses were also performed using Biolog Microbial ID System according to the manufacturers' specifications [63].…”

“…The first PCR-RAPD approach was taken by Oprean, when different Saccharomyces and non-Saccharomyces strains, isolated from Sebes , -Apold vineyard, were identified [65]. Relatively recently, apart from using the ITS-RFLP technique of the ITS1-5.8S rDNA-ITS2 region, taking advantage of the restriction enzymes such as Hinf I, HaeIII, CfoI, and MspI, Corbu and Csutak have also employed the RAPD method for the identification of yeast involved in wine spontaneous fermentation [62,63]. In their case, the intraspecific biodiversity (genetic relatedness) of the isolates was detected by analyzing the RAPD profile obtained for each strain and by calculating the similarity index using the Jaccard coefficient (Sij).…”

Promising Probiotic Properties of the Yeasts Isolated from Rabilé, a Traditionally Fermented Beer Produced in Burkina Faso

“…Additionally, S. cerevisiae , the best-known species, has been extensively studied for its valuable probiotic characteristics [ 33 , 34 , 35 ]. Currently, non- Saccharomyces species have been studied for their potential probiotic attributes [ 33 , 34 , 35 , 36 , 37 , 38 , 39 ]. Ogunremi et al [ 36 ] have isolated new yeast strains from some traditional cereal-based fermented products from Nigeria, such as Candida tropicalis , Issatchenkia orientalis , Pichia kudriavzevii , and Pichia kluyveri , and characterized them as starter cultures with multifunctional potentials to produce cereal-based probiotic products.…”