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ORIGINAL RESEARCH article

Front. Microbiol.
Sec. Antimicrobials, Resistance and Chemotherapy
Volume 15 - 2024 | doi: 10.3389/fmicb.2024.1475172

Identification and characterization of a novel chromosome-encoded aminoglycoside O-nucleotidyltransferase gene, ant(9)-Id, in Providencia sp. TYF-12 isolated from the marine fish intestine

Provisionally accepted
Yan Yu Yan Yu 1*Runzhi Zhang Runzhi Zhang 2*Wei Pan Wei Pan 3*Susu Chen Susu Chen 4*Junjun Wang Junjun Wang 4*Junwan Lu Junwan Lu 5Qiyu Bao Qiyu Bao 1,4,5Yunliang Hu Yunliang Hu 1*Pengfei Jiang Pengfei Jiang 2*DaWei Huang DaWei Huang 3*
  • 1 Institute of Biomedical Informatics/School of Laboratory Medicine and Life Sciences, Wenzhou Medical University, Wenzhou 325035, China
  • 2 Institute of Molecular Virology and Immunology, Department of Microbiology and Immunology, School of Basic Medical Sciences, Wenzhou Medical University, Wenzhou 325035, China
  • 3 Department of Laboratory Sciences, The People’s Hospital of Yuhuan, Yuhuan 317600, China
  • 4 Department of Laboratory Sciences, Pingyang Hospital of Wenzhou Medical University, Pingyang 325400, China
  • 5 Medical Molecular Biology Laboratory, School of Medicine, Jinhua University of Vocational Technology, Jinhua 321000, China

The final, formatted version of the article will be published soon.

    The mechanisms underlying the resistance of the genus Providencia to aminoglycosides are complex, which poses a challenge for the efficient treatment of infectious diseases caused by these pathogens. To help clinicians treat infections more effectively, a more comprehensive understanding of antibiotic resistance mechanisms is urgently needed.Plates were streaked to isolate bacteria from the intestinal contents of fish.The standard agar dilution method was used to determine the minimum inhibitory concentrations (MICs) of the antimicrobial agents. Molecular cloning was carried out to study the function of the novel antibiotic inactivation gene ant( 9)-Id. The kinetic parameters of ANT(9)-Id were measured by a SpectraMax multifunctional microplate reader. Whole-genome sequencing and bioinformatic analysis were conducted to elucidate the sequence structure and evolutionary relationships of similar genes.The novel aminoglycoside O-nucleotidyltransferase gene ant( 9)-Id was encoded on the chromosome of a species-unclassified isolate designated Providencia sp. TYF-12, which was isolated from the intestine of a marine fish. Among the 11 aminoglycosides tested, ant(9)-Id was resistant to only spectinomycin. The MIC of spectinomycin for the recombinant strain carrying ant(9)-Id (pUCP20-ant(9)-Id/DH5α) increased 64-fold compared with that of the control strain (pUCP20/DH5ɑ). ANT(9)-Id shares the highest amino acid (aa) identity of 46.70% with the known drug resistance enzyme ANT(9)-Ic. Consistent with the MIC results, ANT(9)-Id showed high affinity and catalytic efficiency for spectinomycin, with a Km of 8.94 ± 2.50 μM and a kcat/Km of 26.15 μM -1 •s -1 . This novel resistance gene and its close homologs are conserved in Providencia strains from various sources, including some of clinical significance. No mobile genetic elements (MGEs) surrounding the ant(9)-Id(-like) genes were identified.This work revealed and characterized a novel spectinomycin resistance gene, ant(9)-Id, along with its biological features. Identifying novel resistance genes in pathogens can assist in rational medication use and the identification of additional antimicrobial resistance mechanisms in microbial populations.

    Keywords: Providencia, resistance gene, ant(9)-Id, aminoglycoside O-nucleotidyltransferase, Kinetic parameter

    Received: 03 Aug 2024; Accepted: 19 Nov 2024.

    Copyright: © 2024 Yu, Zhang, Pan, Chen, Wang, Lu, Bao, Hu, Jiang and Huang. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

    * Correspondence:
    Yan Yu, Institute of Biomedical Informatics/School of Laboratory Medicine and Life Sciences, Wenzhou Medical University, Wenzhou 325035, China
    Runzhi Zhang, Institute of Molecular Virology and Immunology, Department of Microbiology and Immunology, School of Basic Medical Sciences, Wenzhou Medical University, Wenzhou 325035, China
    Wei Pan, Department of Laboratory Sciences, The People’s Hospital of Yuhuan, Yuhuan 317600, China
    Susu Chen, Department of Laboratory Sciences, Pingyang Hospital of Wenzhou Medical University, Pingyang 325400, China
    Junjun Wang, Department of Laboratory Sciences, Pingyang Hospital of Wenzhou Medical University, Pingyang 325400, China
    Yunliang Hu, Institute of Biomedical Informatics/School of Laboratory Medicine and Life Sciences, Wenzhou Medical University, Wenzhou 325035, China
    Pengfei Jiang, Institute of Molecular Virology and Immunology, Department of Microbiology and Immunology, School of Basic Medical Sciences, Wenzhou Medical University, Wenzhou 325035, China
    DaWei Huang, Department of Laboratory Sciences, The People’s Hospital of Yuhuan, Yuhuan 317600, China

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