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.

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.