The authors have recently developed the overexpression profiling ADOPT method. In the ADOPT method, yeast strains overexpressing most of the genes in the budding yeast Saccharomyces cerevisiae genome are mixed and competitively cultured, and the genes overexpressed in the enriched strains are systematically identified. Furthermore, the identified genes can be used to identify bottleneck factors that are necessary but lacking for growth of S. cerevisiae under given conditions. In our previous studies, we have identified bottlenecks in artificially created stress environments in the laboratory, but in this study, we used grape juice for winemaking as an example to see if industrial bottlenecks can be identified. ADOPT experiments with sulfite-added grape juice used in conventional winemaking resulted in a strong enrichment of strains overexpressing the sulfite pump SSU1 and its transcription factor FZF1. Since enhancement of SSU1 function is known to occur in wine yeast acclimation, ADOPT was also shown to be useful in the search for industrial bottlenecks. On the other hand, no genes were strongly enriched by ADOPT in grape juice without sulfite addition, suggesting that grape juice is a balanced medium with few bottlenecks for S. cerevisiae growth.