Changes in Bacterial Populations and Their Metabolism over 90 Sequential Cultures on Wheat-Based Thin Stillage

Wheat-based thin stillage (W-TS) is a liquid co-product of wheat fermentation for ethanol production, which typically contains substantial amounts of glycerol. Two-stage fermentation, via endemic microorganisms, can be used in processes to convert this compound to more valuable products and simplify the enrichment process through the clarification of the medium and concentration of particles as a protein-rich concentrate. We recultured bacteria 90 times (72 h at 37 °C) on fresh W-TS to determine the stability of the culture and metabolic processes. Next-generation sequencing of W-TS revealed the presence of a predominant Lactobacillus community that rapidly displaced competing microorganisms (e.g., Pediococcus) in subsequent fermentations. These organisms produced bacteriocins (e.g., helveticin J, interpreted through the presence of bacteriocin genes) and acidified the fermentation broth (through the production of succinic acid: 1.7 g/L, lactic acid: 1.8 g/L, and acetic acid: 4.1 g/L). Furthermore, the microbial community produced cobalamin (inferred through sequencing) and converted glycerol (10 g/L reduced to 3.5 g/L after 72 h) to 1,3-propanediol (6.1 g/L after 72 h). Altogether, Lactobacilli were identified as the predominant endemic microorganisms in W-TS after the first 10 cultures. The community was stable and provided a novel approach to increase the value of organic solutes in W-TS.