Changes in the Polyphenolic Profile and Antioxidant Activity of Wheat Bread after Incorporating Quinoa Flour.

Quinoa is a trend and a promising functional food ingredient. Following previous research into the impact of incorporating quinoa flour on the polyphenol content and antioxidant activity of bread, this study aimed to bridge an existing gap about the qualitative and quantitative polyphenolic profiles of such bread. The UPLC-MS/MS analysis showed that quinoa bread, made with 25% quinoa flour of a black variety, presented more compounds than refined-wheat bread, and levels were remarkably higher in many cases. Consequently, the quinoa bread presented clearly improved polyphenolic content than the wheat bread (12.8-fold higher considering the sum of extractable and hydrolyzable polyphenols), as supported by greater antioxidant activity (around 3-fold). The predominant compounds in the extractable fraction of quinoa bread were p-hydroxybenzoic acid and quercetin (50- and 64-fold higher than in wheat bread, respectively) and rutin (not detected in wheat bread), while ferulic and sinapic acids were the most abundant compounds in the hydrolyzable fraction (7.6- and 13-fold higher than in wheat bread, respectively). The bread-making impact was estimated, and a different behavior for phenolic acids and flavonoids was observed. Extractable phenolic acids were the compounds that decreased the most; only 2 of 12 compounds were enhanced (p-hydroxybenozoic and rosmarinic acid with increments of 64% and 435%, respectively). Flavonoids were generally less affected, and their concentrations considerably rose after the bread-making process (7 of the 13 compounds were enhanced in the extractable fraction) with especially noticeably increases in some cases; e.g., apigenin (876%), kaempferol (1304%), luteolin (580%) and quercetin (4762%). Increments in some extractable flavonoids might be explained as a consequence of the release of the corresponding hydrolyzable forms. The present study provides new information on the suitability of quinoa-containing bread as a suitable vehicle to enhance polyphenols intake and, hence, the antioxidant activity in daily diets.

Proteomic Analysis of Saccharomyces cerevisiae Response to Oxidative Stress Mediated by Cocoa Polyphenols Extract.

The present study addressed the protective effects against oxidative stress (OS) of a cocoa powder extract (CPEX) on the protein expression profile of S. cerevisiae. A proteomic analysis was performed after culture preincubation with CPEX either without stress (-OS) or under stress conditions (+OS) (5 mM of H2O2). LC-MS/MS identified 33 differentially expressed proteins (-OS: 14, +OS: 19) that were included By Gene Ontology analysis in biological processes: biosynthesis of amino acids, carbohydrate metabolism and reactive oxygen species metabolic process. In a gene-knockout strains study, eight proteins were identified as putative candidates for being involved in the protective mechanism of cocoa polyphenols against OS induced by H2O2. CPEX was able to exert its antioxidant activity in yeast mainly through the regulation of: (a) amino acids metabolism proteins by modulating the production of molecules with known antioxidant roles; (b) stress-responsive protein Yhb1, but we were unable to fully understand its down-regulation; (c) protein Prb1, which can act by clipping Histone H3 N-terminal tails that are related to cellular resistance to DNA damaging agents.

Effect of Incorporating White, Red or Black Quinoa Flours on Free and Bound Polyphenol Content, Antioxidant Activity and Colour of Bread.

Interest in quinoa as a functional food ingredient is currently emerging. The flours from white, red and black quinoa seeds were analysed in terms of total polyphenol content and antioxidant activity. They were incorporated at 25% on flour basis into the bread dough formula to evaluate their potential to improve the functional properties of wheat breads. The contribution of extractable polyphenols (soluble forms) and the largely unexplored hydrolysable polyphenols (bound forms that can be found in the residues of the former) were taken into account to reflect a realistic health-promoting potential of breads. The red and black quinoa varieties stood out compared to wheat flour, with about double the polyphenol content and up to 4.7-fold increments in antioxidant activity when considering the sum of extractable and hydrolysable polyphenols. The red and black flours were equally effective in intensifying the antioxidant properties of bread despite the baking process (between 2- and 3-fold). They produced significant changes in the parameters that describe crust and crumb colour (L*, a*, b*). A clear darkening was observed compared to the control bread, an appealing attribute for lovers of unconventional and natural products. According to our results, the flours from the coloured quinoa seeds could be considered interesting antioxidant sources and be applied as natural ingredients in bread-making; new, promising and valuable unconventional products for consumers and producers could be developed.

Evaluation of the Ability of Polyphenol Extracts of Cocoa and Red Grape to Promote the Antioxidant Response in Yeast Using a Rapid Multiwell Assay.

Saccharomyces cerevisiae has been used as a model organism to study the capacity of cocoa and red grape extracts to trigger an antioxidant response. A methodology adapted to microtiter plates has been developed to monitor yeast growth after culture preincubation with food ingredients and exposure to oxidative stress by hydrogen peroxide and menadione. This methodology proved effective in measuring the ability of cocoa and red grape extracts to promote an antioxidant response in yeast, and also the prospect of conducting dose-response studies. Additionally, the method has proven useful to perform studies with mutant strains lacking genes that may be related to the mechanism of action underlying the antioxidant properties. Thus, in a single assay, it is possible to elucidate the sensitivity of strains to oxidative stress, the ability of an ingredient to promote an antioxidant response, and the possible implication of certain genes. Results of assays using strain hst3Δ showed that the antioxidant protection provided by exposure to cocoa and red grape extracts was not present in the strain lacking gene HST3 when H2 O2 and menadione were used as oxidizing agents. This effect was previously reported for cocoa extract only, with H2 O2 as stressor. Moreover, the results showed that the mutant strain hst3Δ is more resistant to menadione and H2 O2 in the absence of preincubation with cocoa and red grape extract, hinting at the possible implication of sirtuin Hst3 in the antioxidant cellular response.

Differences in activation of MAP kinases and variability in the polyglutamine tract of Slt2 in clinical and non-clinical isolates of Saccharomyces cerevisiae.

The concept of Saccharomyces cerevisiae as an emerging opportunistic pathogen is relatively new and it is due to an increasing number of human infections during the past 20 years. There are still few studies addressing the mechanisms of infection of this yeast species. Moreover, little is known about how S. cerevisiae cells sense and respond to the harsh conditions imposed by the host, and whether this response is different between clinical isolates and non-pathogenic strains. In this regard, mitogen-activated protein kinase (MAPK) pathways constitute one of the major mechanisms for controlling transcriptional responses and, in some cases, virulence in fungi. Here we show differences among clinical and non-clinical isolates of S. cerevisiae in the level of activation of the MAPKs Kss1, which controls pseudohyphal and invasive growth, and Slt2, which is required for maintaining the integrity of the cell wall under stress conditions and in the absence of stimulating conditions. Moreover, we report for the first time the existence of length variability in SLT2 alleles of strains with a clinical origin. This is due to the expansion in the number of glutamine-encoding triplets in the microsatellite region coding for the polyglutamine (poly-Q) tract of this gene, which range from 12 to more than 38 repetitions. We suggest that this variability may influence biological features of the Slt2 protein, allowing it to adapt swiftly in order to survive in unusual environments.

Food and probiotic strains from the Saccharomyces cerevisiae species as a possible origin of human systemic infections.

We report four cases of blood cultures testing positive for yeast strains belonging to the species Saccharomyces cerevisiae. Using molecular techniques, RFLP of mtDNA and delta-PCR amplification, we show the association of two of the isolates with non-clinical strains. Specifically, with two commercial bread-making strains and the therapeutic S. boulardii strain. The association of S. boulardii with cases of fungemia has been reported previously. Nevertheless, this is the first time that a baker's yeast has been isolated from blood.

Molecular evolution in yeast of biotechnological interest.

The importance of yeast in the food and beverage industries was only realized about 1860, when the role of these organisms in food manufacture became evident. Since they grow on a wide range of substrates and can tolerate extreme physicochemical conditions, yeasts, especially the genera Saccharomyces and Kluyveromyces, have been applied to many industrial processes, Industrial strains of these genera are highly specialized organisms that have evolved to utilize a range of environments and ecological niches to their full potential. This adaptation is called "domestication". This review describes the phylogenetic relationships among Saccharomyces and Kluyveromyces species and the different mechanisms involved in the adaptive evolution of industrial yeast strains.