Characterization of yeast population from unstudied natural sources in La Mancha region.

AIMS: This study aims to identify the yeast species and strains which entitled an unstudied area of Spain and evaluate the yeast species diversity richness and the genetic variety. METHODS AND RESULTS: A total of 702 yeasts were isolated from different environments in a central Spanish region (La Mancha) with diverse sources of origin (food, animals, flowers and environmental sources) during spring season. Thanks to the analysis carried out by the PCR-RFLP technique and sequencing, 35 species were identified. A neighbour-joining phylogenetic tree was created based on D1/D2 sequences. Moreover 330 strains were determined by PCR-RAPD and their profiles were analysed using the bioinformatics programme BioNumerics 7·6. The Simpson's index (D) and the genetic diversity percentage were calculated with the aim of studying the richness of the species in each environment and the genetic variety in each species. CONCLUSIONS: This study has permitted to know that the majority of the species found was Diutina rugosa while the most ubiquitous was Rhodotorula mucilaginosa which expose the dispersion capability of this species. The diversity parameters has revealed that the highest species richness was associated to environmental samples and the highest genetic variety was presented in those species with better dispersion capability or a smaller number of isolates. SIGNIFICANCE AND IMPACT OF THE STUDY: This study permits to better understand the yeast communities in La Mancha region which gives a value the microbial potential of this region.

Biotechnological application of yeasts in food science: Starter cultures, probiotics and enzyme production.

This review is an account of experiences of two research teams (from Italy to Spain); the leading idea is the following: yeasts represent valuable sources in food science and microbiology and are a kind of food factories, because of the potentiality of whole cells or for their produced compounds. This review covers three major areas: the first section addresses the role of yeasts as starter cultures with a special focus on wine. The second section is an update on probiotic yeasts. Finally, the focus of the last section is on enzymes produced by yeasts, with a short description of the removal of mycotoxin.

Co-inoculation of different Saccharomyces cerevisiae strains and influence on volatile composition of wines.

Wine is the result of the performance of different yeast strains throughout the fermentation in both spontaneous and inoculated processes. 22 Saccharomyces cerevisiae strains were characterized by microsatellite fingerprinting, selecting 6 of them to formulate S. cerevisiae mixed cultures. The aim of this study was to ascertain a potential benefit to use mixed cultures to improve wine quality. For this purpose yeasts behavior was studied during co-inoculated fermentations. Aromatic composition of the wines obtained was analyzed, and despite the fact that only one strain dominated at the end of the process, co-cultures released different concentrations of major volatile compounds than single strains, especially higher alcohols and acetaldehydes. Nevertheless, no significant differences were found in the type and quantity of the amino acids assimilated. This study demonstrates that the final wine composition may be modulated and enhanced by using suitable combinations of yeast strains.

Amino acid uptake by wild and commercial yeasts in single fermentations and co-fermentations.

Musts require nitrogen-containing compounds in order to ensure yeast development. This study examined the nitrogen-nutrient requirements of two commercial yeasts and three wild strains isolated from inoculated fermentations. The results showed that wild strains generally consumed lower amounts of amino acids than commercial yeasts. Most amino acids were assimilated during the exponential growth phase; only a few - including asparagine and histidine - were metabolized until the end of fermentation. The study also sought to determine whether industrial drying affected yeast nitrogen requirements.

Optimization of a rapid method for studying the cellular location of beta-glucosidase activity in wine yeasts.

AIMS: To improve a method for determining beta-glucosidase activity and to apply it in yeasts isolated from wine ecosystems from "La Mancha" region and to know its cellular location. METHODS AND RESULTS: A total of 82 wine yeasts were identified (PCR/RFLP) and evaluated for their beta-glucosidase activity. First, they were qualitatively evaluated by growth on YNB cellobiose, the activity was quantified using different culture media, under aerobic and anaerobic conditions and cells after 24-72 h of growth. To study the location activity, five fractions were obtained (supernatant, whole cell, cell wall, cytosol and cell membrane). The enzymatic assays were optimized, being: growth in YP cellobiose for 72 h in aeration conditions and, after cell removing, enzyme analysis with 128 g l(-1) of cellobiose as substrate, for 30 min at 30 degrees C. The genus that displayed the greatest activity were Pichia, Hanseniaspora and Rhodotorula, and the activity was intracellular. CONCLUSIONS: The study showed that beta-glucosidase activity was induced by the carbon source and was aerobic dependent. The non-Saccharomyces species displayed the greatest activity, which was intracellular and strain-dependent. SIGNIFICANCE AND IMPACT OF THE STUDY: This study developed a reliable method for screening beta-glucosidase activity in yeasts isolated from wine ecosystems. This activity is very important in the release of monoterpenols from glycoside precursors for the enhancement of wine aromas.