In this study, through quantitative detection of key substances and enzyme activities, an integrated analysis of 16S rRNA sequencing and metatranscriptomics revealed the mechanisms by which salt and oil influence the biotransformation process during anaerobic digestion (AD). The results demonstrated that a salt concentration of 6 g/L promoted lipid metabolism and hydrogenotrophic methanogenesis, while inhibiting the acetoclastic pathway. An oil concentration of 5 g/L facilitated the expression of key enzyme-encoding genes involved in β-oxidation of long-chain fatty acids, transcription, and acetoclastic methanogenesis. It also promoted the enrichment of syntrophic propionate/butyrate oxidation bacteria (Syntrophomonas and DMER64). Salt/oil co-addition enhanced the expression of genes related to glucose metabolism, amino acid metabolism, organic acid synthesis, and quorum sensing. Furthermore, salt/oil co-addition inhibited the secretion of key enzymes related to methanogens by impeding the transcription process. Collectively, these findings provide systematic insights into how salt and oil affect the biochemical metabolic mechanisms of AD.
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