Frontiers in cellular and infection microbiology · 2026
Discovery and characterization of a novel chromosomally encoded aminoglycoside O-nucleotidyltransferase gene, designated ant(9)-Ie, in a strain of Providencia.
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Huang Lulu, Yang Zhigang, Wu Xiaolan, Hu Ruxi, Hu Xiaofang, Chen Wenjie, Mao Xuanjiao, Chen Susu, Li Zhengqiao, Gong Jie, Lu Junwan, Bao Qiyu, Song Chunhan, Xu Teng, Zhang Hongqin
Аннотация
BACKGROUND: The complex aminoglycoside resistance mechanisms observed within the genus Providencia present formidable clinical challenges to the effective management of the infectious diseases caused by these species. In-depth research into the mechanisms of bacterial resistance to antimicrobials must be conducted to strengthen the response capabilities of healthcare facilities for infection prevention and control.
METHODS: Bacteria from wastewater from an animal farm were isolated by plate streaking. The minimum inhibitory concentrations (MICs) of the antimicrobials were determined using the standard agar dilution method. The function of the novel resistance gene ant(9)-Ie was elucidated using molecular cloning technology. ANT(9)-Ie was heterologously expressed, and its kinetic parameters were determined. The biological characteristics of the genome were systematically analyzed through whole-genome sequencing and a comparative genomic analysis, and a phylogenetic analysis of the genetic background for resistance gene-related sequences was performed.
RESULTS: In Providencia rettgeri P17, a novel resistance gene, designated ant(9)-Ie, conferring resistance against spectinomycin was identified. A comparative analysis revealed that among all the functionally characterized resistance proteins, ANT(9)-Ie had the highest amino acid (aa) sequence similarity to ANT(9)-Id (58.67%). Compared with the control strain E. coli DH5α (DH5α), the recombinant strain harboring ant(9)-Ie (pMD19-ant(9)-Ie/DH5α) showed a 64-fold increase in the MIC of spectinomycin. The results of the kinetic analysis of ANT(9)-Ie were generally consistent with the MIC for the cloned ant(9)-Ie, with a high spectinomycin catalytic efficiency [k cat/K m, (8.22 ± 1.24) × 102 M-1·s-1). No new features were observed in the domains or motifs of the protein. The ant(9)-Ie homologous genes were found only in Providencia strains and exhibited a relatively conserved genetic environment. The Providencia strains whose genomes were available in the NCBI database were isolated from different sources, most of which were from human clinical specimens.
CONCLUSIONS: In this study, a novel spectinomycin resistance gene, ant(9)-Ie, was identified, and its biological features were characterized. The identification of this novel resistance gene from Providencia might aid in the effective clinical treatment of infections caused by bacteria carrying this resistance gene in the future and paves the way for further research into the complexity of resistance mechanisms within microbial populations.
METHODS: Bacteria from wastewater from an animal farm were isolated by plate streaking. The minimum inhibitory concentrations (MICs) of the antimicrobials were determined using the standard agar dilution method. The function of the novel resistance gene ant(9)-Ie was elucidated using molecular cloning technology. ANT(9)-Ie was heterologously expressed, and its kinetic parameters were determined. The biological characteristics of the genome were systematically analyzed through whole-genome sequencing and a comparative genomic analysis, and a phylogenetic analysis of the genetic background for resistance gene-related sequences was performed.
RESULTS: In Providencia rettgeri P17, a novel resistance gene, designated ant(9)-Ie, conferring resistance against spectinomycin was identified. A comparative analysis revealed that among all the functionally characterized resistance proteins, ANT(9)-Ie had the highest amino acid (aa) sequence similarity to ANT(9)-Id (58.67%). Compared with the control strain E. coli DH5α (DH5α), the recombinant strain harboring ant(9)-Ie (pMD19-ant(9)-Ie/DH5α) showed a 64-fold increase in the MIC of spectinomycin. The results of the kinetic analysis of ANT(9)-Ie were generally consistent with the MIC for the cloned ant(9)-Ie, with a high spectinomycin catalytic efficiency [k cat/K m, (8.22 ± 1.24) × 102 M-1·s-1). No new features were observed in the domains or motifs of the protein. The ant(9)-Ie homologous genes were found only in Providencia strains and exhibited a relatively conserved genetic environment. The Providencia strains whose genomes were available in the NCBI database were isolated from different sources, most of which were from human clinical specimens.
CONCLUSIONS: In this study, a novel spectinomycin resistance gene, ant(9)-Ie, was identified, and its biological features were characterized. The identification of this novel resistance gene from Providencia might aid in the effective clinical treatment of infections caused by bacteria carrying this resistance gene in the future and paves the way for further research into the complexity of resistance mechanisms within microbial populations.