This study used a rumen simulation technique to evaluate the effects of soaking of by-product-rich concentrate (BPC) in 5% lactic acid (LAC; vol/vol) on the rumen microbiota, predicted metagenome, fermentation characteristics, and nutrient degradation without or with supplemented P. The diet was supplemented with 1.6 g of P in the form of monocalcium phosphate per kilogram of dry matter in addition to 284 mg of inorganic P/d per fermentor via artificial saliva. Fermentor fluid was collected for analyses of short-chain fatty acids, fermentation gases, redox potential, and microbiota and feed residues for calculation of nutrient degradation. The microbiota composition was assessed using paired-end Illumina (Illumina Inc., San Diego, CA) MiSeq sequencing of the V3 to V5 region of the 16S rRNA gene. Soaking in LAC reduced the contents of crude protein, neutral and acid detergent fibers, and organic matter fractions as well as ash and P content of the BPC. Both the LAC treatment of BPC and the inorganic P modified the relative bacterial abundances mainly within the predominant orders Bacteroidales and Clostridiales. Supervised DIABLO N-integration networking supported that operational taxonomic units related to BS11, Ruminococcaceae, Christensenellaceae, Eubacterium, and Selenomonas were the most discriminant for the LAC-treated BPC, whereas other operational taxonomic units related to BS11, RFN20, Ruminococcus, and Succiniclasticum were best correlated with the inorganic P supplementation. Integration networking also showed that carbohydrate and pyruvate metabolism, biosynthesis of unsaturated fatty acids, and degradation of several xenobiotics were stimulated by the LAC treatment of BPC. Those data supported the enhanced fermentation activity as indicated by increased total short-chain fatty acid concentration, especially propionate and butyrate, and methane, but decreased ruminal crude protein degradation, with the LAC-treated compared with control-treated BPC. In contrast, despite an increased abundance of imputed functions, such as inositol phosphate metabolism, phosphatidylinositol signaling, and fructose and mannose metabolism, the reduced abundance of the imputed Kyoto Encyclopedia of Genes and Genomes pathway "transcription machinery" as well as the decrease in total short-chain fatty acids and nutrient degradation indicated reduced bacterial metabolic activity with the inorganic P supplementation. In conclusion, soaking of BPC in LAC may favor the proliferation of certain fibrolytic bacterial taxa and stimulate their metabolic activity, whereas the supplemented P to a diet already meeting ruminal P needs may impair ruminal nutrient utilization.