When confronted with environmental stress, yeast cell reacts, among others, by modifying the expression of specific genes. In this study, gene expression was analyzed via RT-qPCR to quantify the oxidative stress of Saccharomyces pastorianus during yeast propagation as a reaction to different aeration levels. Target genes were identified, and a reference gene system was developed. Fermentation experiments were conducted in shaking flasks, applying different shaking speeds to generate various aeration efficiencies. The cells were sampled at different propagation stages and, additionally to the expression study, analyzed by flow cytometry after staining with dihydroethidium (DHE) to quantify reactive oxygen species (ROS) inside the cells. The results indicate that high oxygen fermentation conditions led to an increased expression of the catalase-A gene CTA1 during propagation. Furthermore, the determination of cell internal ROS shows increasing oxidative stress over the process in accordance with the RT-qPCR measurements.

Consumer health concerns and regulatory policies lead to a growing demand for sugar-sweetened beverage alternatives. A reduced energy content can be achieved using artificial sweeteners, which often also convey a metallic or bitter off-flavor. Therefore, the alteration of sweetness perception and masking of potential off-flavors are paramount for improving sweet beverages. Trigeminal stimuli, such as capsaicin (spicy) or menthol (cooling), have been used to influence taste perception in food items, although their use in beverages has not yet been systematically investigated. Here, the influence of menthol on sweetness perception in an aqueous solution is examined both on the sensory and psychophysiological level. The addition of menthol had no sensory effect on sweetness perception; however, psychophysiological measurements suggest a boost in the physiological response to cold perception through the addition of sugar. Moreover, menthol addition shifted the recognition threshold of unpleasant bitterness of the sweetener acesulfame-K from 21.35 to 36.93 mg/L, masking the off-flavor. These findings illuminate the complexity of trigeminal perception influences on taste. Further investigation of these effects can render trigeminal stimuli an effective tool to enhance beverage aroma and flavor.

Targeted induced gene expression for industrial fermentation processes in food and beverage production could fulfill future demands. To avoid metabolic burden and disturbances owing to the fermentation procedure, induced gene expression is necessary for combating stress, such as that caused by temperature shifts that occur during the transition from fermentation to maturation in the brewing process. The aim of this study was to target gene expression in industrial yeast using stress-responsive promoters and homologues of the selection marker SMR1. Self-cloning strains of the industrial brewing yeast Saccharomyces pastorianus TUM 34/70 were constructed to overexpress the alcohol acetyltransferase (ATF1) gene under the control of inducible promoters SSA3, HSP104 and UBI4. Transcription analysis shows the highest induction after 72 h of shock situation for HSP104 with 1.3-fold and UBI4 with 2.2-fold. Further, at the end of shock situation the concentrations of ethyl acetate were 1.2- and 1.3-fold higher than the wild type for HSP104 and UBI4, respectively. In addition, the influence of the final temperature and temporal sequence of temperature shock to 4°C had a major impact on expression patterns. Therefore, these data show that temperature-induced gene expression of self-cloning industrial yeast could be an option for optimization of the beverage fermentation.