{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,25]],"date-time":"2026-09-25T21:44:40Z","timestamp":1790372680753,"version":"4.1.0"},"reference-count":42,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2024,7,30]],"date-time":"2024-07-30T00:00:00Z","timestamp":1722297600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Cell. Infect. Microbiol."],"abstract":"<jats:sec><jats:title>Background<\/jats:title><jats:p>Aminoglycoside-modifying enzymes (AMEs) play an essential role in bacterial resistance to aminoglycoside antimicrobials. With the development of sequencing techniques, more bacterial genomes have been sequenced, which has aided in the discovery of an increasing number of novel resistance mechanisms.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>The bacterial species was identified by 16S rRNA gene homology and average nucleotide identity (ANI) analyses. The minimum inhibitory concentration (MIC) of each antimicrobial was determined by the agar dilution method. The protein was expressed with the pCold I vector in <jats:italic>E. coli<\/jats:italic> BL21, and enzyme kinetic parameters were examined. The whole-genome sequence of the bacterium was obtained via the Illumina and PacBio sequencing platforms. Reconstruction of the phylogenetic tree, identification of conserved functional residues, and gene context analysis were performed using the corresponding bioinformatic techniques.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>A novel aminoglycoside resistance gene, designated <jats:italic>aph(3\u2019)-Ie<\/jats:italic>, which confers resistance to ribostamycin, kanamycin, sisomicin and paromomycin, was identified in the chromosome of the animal bacterium <jats:italic>Citrobacter gillenii<\/jats:italic> DW61, which exhibited a multidrug resistance phenotype. APH(3\u2019)-Ie showed the highest amino acid identity of 74.90% with the functionally characterized enzyme APH(3\u2019)-Ia. Enzyme kinetics analysis demonstrated that it had phosphorylation activity toward four aminoglycoside substrates, exhibiting the highest affinity (<jats:italic>K<\/jats:italic><jats:sub>m<\/jats:sub>, 4.22 \u00b1 0.88 \u00b5M) and the highest catalytic efficiency [<jats:italic>k<\/jats:italic><jats:sub>cat<\/jats:sub>\/<jats:italic>K<\/jats:italic><jats:sub>m<\/jats:sub>, (32.27 \u00b1 8.14) \u00d7 10<jats:sup>4<\/jats:sup>] for ribomycin. Similar to the other APH(3\u2019) proteins, APH(3\u2019)-Ie contained all the conserved functional sites of the APH family. The <jats:italic>aph(3\u2019)-Ie<\/jats:italic> homologous genes were present in <jats:italic>C. gillenii<\/jats:italic> isolates from different sources, including some of clinical significance.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusion<\/jats:title><jats:p>In this work, a novel chromosomal aminoglycoside resistance gene, designated <jats:italic>aph(3\u2019)-Ie<\/jats:italic>, conferring resistance to aminoglycoside antimicrobials, was identified in a rabbit isolate <jats:italic>C. gillenii<\/jats:italic> DW61. The elucidation of the novel resistance mechanism will aid in the effective treatment of infections caused by pathogens carrying such resistance genes.<\/jats:p><\/jats:sec>","DOI":"10.3389\/fcimb.2024.1435123","type":"journal-article","created":{"date-parts":[[2024,7,30]],"date-time":"2024-07-30T15:09:18Z","timestamp":1722352158000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":8,"title":["APH(3\u2019)-Ie, an aminoglycoside-modifying enzyme discovered in a rabbit-derived Citrobacter gillenii isolate"],"prefix":"10.3389","volume":"14","author":[{"given":"Naru","family":"Lin","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuning","family":"Sha","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Guozhi","family":"Zhang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chunhan","family":"Song","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuan","family":"Zhang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jingxuan","family":"Zhao","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dawei","family":"Huang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Junwan","family":"Lu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Qiyu","family":"Bao","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wei","family":"Pan","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2024,7,30]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"D690","DOI":"10.1093\/nar\/gkac920","article-title":"CARD 2023: expanded curation, support for machine learning, and resistome prediction at the Comprehensive Antibiotic Resistance Database","volume":"51","author":"Alcock","year":"2023","journal-title":"Nucleic Acids Res."},{"key":"B2","doi-asserted-by":"publisher","DOI":"10.1099\/mgen.0.000923","article-title":"Multidrug-resistant toxigenic Corynebacterium diphtheriae sublineage 453 with two novel resistance genomic islands","volume":"9","author":"Arcari","year":"2023","journal-title":"Microbial Genomics"},{"key":"B3","doi-asserted-by":"publisher","DOI":"10.1002\/chem.202200883","article-title":"Late-stage modification of aminoglycoside antibiotics overcomes bacterial resistance mediated by APH(3') kinases","volume":"28","author":"Bastian","year":"2022","journal-title":"Chem. 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