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Microbial degradation of salicylate has been well studied and salicylate hydroxylases play essential roles in linking the peripheral and ring\u2010cleavage catabolic pathways. The direct hydroxylation of salicylate catalyzed by salicylate\u20101\u2010hydroxylase or salicylate\u20105\u2010hydroxylase has been well studied. However, the CoA mediated salicylate 5\u2010hydroxylation pathway has not been characterized in detail. Here, we elucidate the molecular mechanism of the reaction in the conversion of salicylate to gentisate in the carbaryl\u2010degrading strain <jats:italic>Rhizobium<\/jats:italic> sp. X9. Three enzymes (salicylyl\u2010CoA ligase CehG, salicylyl\u2010CoA hydroxylase CehH and gentisyl\u2010CoA thioesterase CehI) catalyzed the conversion of salicylate to gentisate via a route, including CoA thioester formation, hydroxylation and thioester hydrolysis. Further analysis indicated that genes <jats:italic>cehGHI<\/jats:italic> are also distributed in other bacteria from terrestrial environment and marine sediments. These genomic evidences highlight the role of this salicylate degradation pathway in the carbon cycle of soil organic compounds and marine sediments. Our findings of this three\u2010step strategy enhanced the current understanding of CoA mediated degradation of salicylate.<\/jats:p>","DOI":"10.1111\/mmi.14771","type":"journal-article","created":{"date-parts":[[2021,6,14]],"date-time":"2021-06-14T08:23:26Z","timestamp":1623659006000},"page":"783-793","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":8,"title":["Unveiling the CoA mediated salicylate catabolic mechanism in <i>Rhizobium<\/i> sp. 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