Regulating the Golgi apparatus sorting of proteinase A to decrease its excretion in Saccharomyces cerevisiae

Beer foam stability, a key factor in evaluating overall beer quality, is influenced by proteinase A (PrA). Actin-severing protein cofilin and Golgi apparatus-localized Ca²⁺ ATPase Pmr1 are involved in protein sorting at the trans-Golgi network (TGN) in yeast Curwin et al. (Mol Biol Cell 23:2327-2338, 2012). To reduce PrA excretion into the beer fermentation broth, we regulated the Golgi apparatus sorting of PrA, thereby facilitating the delivery of more PrA to the vacuoles in the yeast cells. In the present study, the cofilin-coding gene COF1 and the Pmr1-coding gene PMR1 were overexpressed in the parental strain W303-1A and designated as W + COF1 and W + PMR1, respectively. The relative expression levels of COF1 in W + COF1 and PMR1 in W + PMR1 were 5.26- and 19.76-fold higher than those in the parental strain. After increases in the expression levels of cofilin and Pmr1 were confirmed, the PrA activities in the wort broth fermented with W + COF1, W + PMR1, and W303-1A were measured. Results showed that the extracellular PrA activities of W + COF1 and W + PMR1 were decreased by 9.24% and 13.83%, respectively, at the end of the main fermentation compared with that of W303-1A. Meanwhile, no apparent differences were found on the fermentation performance of recombinant and parental strains. The research uncovers an effective strategy for decreasing PrA excretion in Saccharomyces cerevisiae.