Our study shows the possibilities of industrial waste (glycerol and whey lactose) utilization in a microbiological way. In these investigations, propionic acid production by Propionibacterium freudenreichii ssp. shermanii was studied using pure glycerol, crude glycerol, and whey lactose as a carbon sources in culture media. Bacterial growth was carried out in batch fermentation at 20 g/L and 40 g/L initial concentration of carbon sources. Results of conducted experiments show that greater production of propionic acid was achieved with 4% carbon source content in culture media. The highest propionic acid accumulation was 22.57 g/L using whey lactose only in culture media, 24.47 g/L and 24.80 g/L using whey lactose with pure glycerol and whey lactose with crude glycerol as substrates, respectively. It was shown that simultaneous fermentation of whey lactose and pure or crude glycerol did not reduce propionic acid yield compared to whey alone as a substrate in culture media. Moreover, a replacement of 50% of whey lactose by crude or pure glycerol caused lower acetic acid accumulation. Reduction of this by-product should simplify extraction of propionic acid from fermentation broth in the distillation process. These results indicate that industrial wastes could be excellent substrates for propionic acid production.