The Reducing Capacity of Thioredoxin on Oxidized Thiols in Boiled Wort

Free thiol-containing proteins are suggested to work as antioxidants in beer, but the majority of thiols in wort are present in their oxidized form as disulfides and are therefore not active as antioxidants. Thioredoxin, a disulfide-reducing protein, is released into the wort from some yeast strains during fermentation. The capacity of the thioredoxin enzyme system (thioredoxin, thioredoxin reductase, NADPH) to reduce oxidized thiols in boiled wort under fermentation-like conditions was studied. Free thiols were quantitated in boiled wort samples by derivatization with ThioGlo1 and fluorescence detection of thiol-derivatives. When boiled wort was incubated with all components of the thioredoxin system at pH 7.0 and 25 °C for 60 min under anaerobic conditions, the free thiol concentration increased from 25 to 224 μM. At pH values similar to wort (pH 5.7) and beer (pH 4.5), the thioredoxin system was also capable of increasing the free thiol concentration, although with lower efficiency to 187 and 170 μM, respectively. The presence of sulfite, an important antioxidant in beer secreted by the yeast during fermentation, was found to inactivate thioredoxin by sulfitolysis. Reduction of oxidized thiols by the thioredoxin system was therefore only found to be efficient in the absence of sulfite.