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The discovery of these enzymes is often hampered by the limited availability of phage genomes. Herein, we report a new strategy to mine active peptidoglycan hydrolases from bacterial proteomes by lysin-derived antimicrobial peptide-primed screening. As a proof-of-concept, five\n                    <jats:underline>p<\/jats:underline>\n                    eptidoglycan\n                    <jats:underline>h<\/jats:underline>\n                    ydrolases from the\n                    <jats:italic>\n                      <jats:underline>A<\/jats:underline>\n                      cinetobacter\n                      <jats:underline>b<\/jats:underline>\n                      aumannii\n                    <\/jats:italic>\n                    proteome (PHAb7-PHAb11) were identified using PlyF307 lysin-derived peptide as a template. Among them, PHAb10 and PHAb11 showed potent bactericidal activity against multiple pathogens even after treatment at 100\u00b0C for 1 hr, while the other three were thermosensitive. We solved the crystal structures of PHAb8, PHAb10, and PHAb11 and unveiled that hyper-thermostable PHAb10 underwent a unique folding-refolding thermodynamic scheme mediated by a dimer-monomer transition, while thermosensitive PHAb8 formed a monomer. Two mouse models of bacterial infection further demonstrated the safety and efficacy of PHAb10. In conclusion, our antimicrobial peptide-primed strategy provides new clues for the discovery of promising antimicrobial drugs.\n                  <\/jats:p>","DOI":"10.7554\/elife.98266","type":"journal-article","created":{"date-parts":[[2024,7,15]],"date-time":"2024-07-15T20:25:12Z","timestamp":1721075112000},"update-policy":"https:\/\/doi.org\/10.7554\/elife.98266","source":"Crossref","is-referenced-by-count":3,"title":["Dimer-monomer transition defines a hyper-thermostable peptidoglycan hydrolase mined from bacterial proteome by lysin-derived antimicrobial peptide-primed screening"],"prefix":"10.7554","volume":"13","author":[{"given":"Li","family":"Zhang","sequence":"first","affiliation":[{"id":[{"id":"https:\/\/ror.org\/023b72294","id-type":"ROR","asserted-by":"publisher"}],"name":"National Key Laboratory of Agricultural Microbiology & Hubei Hongshan Laboratory, College of Life Science and Technology, Huazhong Agricultural University","place":["Wuhan, China"]},{"name":"College of 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