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Lysine butyrylation (Kbu) is a lysine acylation modification driven by butyrate, a short-chain fatty acid produced by microbial fermentation in the gut. The identification of Kbu has provided new insights into how gut microbial-driven metabolites regulate host protein function and epigenetic process. Kbu has been detected on histone and non-histone proteins, where it contributes to the regulation of gene expression, cellular metabolism, and physiological functions. Growing evidence has linked Kbu to biological processes including cancer progression, inflammation, metabolic disorders, and host-microbiome interactions.