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The PDF file includes:

• Fig. S1. Kinetic curves of enzymatic reaction.
• Fig. S2. MS/MS spectrum of lysine-acylated peptides (YGDEQVKQWR) identified in P. mirabilis.
• Fig. S3. MS/MS spectrum of 2-hydroxyisobutyrylated, acetylated, and succinylated peptides under different incubation conditions.
• Fig. S4. Comparative analysis of Khib, Kac, and Ksucc in P. mirabilis.
• Legends for tables S1 to S5
• Table S6. The list of reagents, bacterial strains, and plasmids used in this paper.

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Other Supplementary Material for this manuscript includes the following:

• Table S1 (Microsoft Excel format). The quantification result of H3K9hib and H3K9 probe enrichment in P. mirabilis protein lysate.
• Table S2 (Microsoft Excel format). The identification result of Kac of P. mirabilis.
• Table S3 (Microsoft Excel format). The identification result of Ksucc of P. mirabilis.
• Table S4 (Microsoft Excel format). The quantification result of 2-hydroisobutyrylated proteomes with CobB KO E. coli and WT E. coli.
• Table S5 (Microsoft Excel format). The PRM result of comparison of the quantities of K326ac and K343hib in T vector and CobB-overexpressing P. mirabilis.