Analysis of the interaction of melatonin with glycolytic proteins in Saccharomyces cerevisiae during alcoholic fermentation
Melatonin is a bioactive compound with antioxidant properties, that has been found in many fermented beverages, such as beer and wine . Indeed, it has been shown that yeast can synthesize melatonin during alcoholic fermentation, although its role inside the cell, as well as the metabolic pathway involved in its synthesis, is still unclear . Recent studies showed that during fermentation, melatonin interacts with different proteins of the glycolytic pathway in both Saccharomyces and non-Saccharomyces yeast, for instance glyceraldehyde 3-phosphate dehydrogenase, pyruvate kinase or enolase . The aim of this study was to deepen in the role of the melatonin within yeast cells, and in the interaction with glycolytic proteins. For that purpose, we performed fermentations with both single and double mutant strains of the different glyceraldehyde 3-phosphate dehydrogenase isoforms (Tdh1, Tdh2 and Tdh3). Melatonin from fermentation samples was analyzed by liquid chromatography mass spectrometry, and proteins bound to melatonin were immunopurified by melatonin IgG-Dynabeads. Intracellular melatonin on the tdhmutant strains during fermentation showed a similar pattern to the wild type strain. Regarding the protein binding to melatonin, in the single and double mutants, we observed that there was only protein binding when the Tdh2 protein was present. Then, we analysed the possible binding sites between Tdh2 and melatonin, using Swissdock and UFCS Chimera programs. The prediction showed that melatonin can form two hydrogen bonds with Tdh2. The results of this study indicate the binding site of melatonin to Tdh2 during fermentation, which could be related to the regulation of yeast carbon metabolism.
This work has been financed by the project PDI2019-108722RB-C33 (MCIN/AEI/10.13039/ 501100011033). SME has had a Martí-Franquès predoctoral grant (2019PMF-PIPF-92).
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Issue: ICGWS 2023