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Isolation and characterization of amoxicillin-resistant bacteria and amoxicillin-induced alteration in its protein profiling and RNA yield

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Abstract

Amoxicillin-resistant bacteria were isolated using selective enrichment procedure. The morphological, biochemical and molecular characterization based on 16S rRNA gene sequencing and phylogenetic analysis of the bacterial strain WA5 confirmed that the strain belongs to the genus Stenotrophomonas. The bacteria were named as Stenotrophomonas sp. strain WA5 (MK110499). Substantial growth was seen in M9 minimal media supplemented with 5 mg L−1 of amoxicillin as a sole source of carbon and energy. RNA yield was also observed to be decreased in the presence of amoxicillin. Amoxicillin (5 mg L−1)-induced alteration is seen on bacterial protein profile and unique polypeptide bands were seen to be induced in the presence of amoxicillin, the bands were subjected to trypsin digestion, and LC–MS/MS analysis showed that the bands belong to the family of DNA-dependent RNA polymerase subunit β (rpoC). Plasmid DNA isolation indicated the presence of antibiotic-resistant genes being harboured by the plasmid.

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Acknowledgements

Financial assistance provided by NASF research grant (project entitled “Bioremediation of chemical contaminants and their complexes present in drainage water with high dynamic flux used for irrigation in urban and peri urban agriculture”), sanction no. NASF/CA-6030/2017-18 is highly acknowledged. The authors Kushneet Kaur Sodhi and Mohit Kumar highly acknowledges the University Grant Commission (UGC), Government of India for providing the stipend.

Funding

This study was supported by the funding agency, National Agricultural Science Fund, Indian Council of Agricultural Research, Delhi, India.

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Correspondence to Dileep Kumar Singh.

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Communicated by Erko Stackebrandt.

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Sodhi, K.K., Kumar, M., Balan, B. et al. Isolation and characterization of amoxicillin-resistant bacteria and amoxicillin-induced alteration in its protein profiling and RNA yield. Arch Microbiol 202, 225–232 (2020). https://doi.org/10.1007/s00203-019-01737-6

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Keywords

  • Amoxicillin
  • 16S rRNA gene
  • Stenotrophomonas
  • LC–MS/MS
  • Bioremediation