Abstract
In bacteria and primitive eukaryotes, sulfonamide antibiotics block the folate pathway by inhibiting dihydropteroate synthase (FolP) that combines para-aminobenzoic acid (pABA) and dihydropterin pyrophosphate (DHPP) to form dihydropteroic acid (DHP), a precursor for tetrahydrofolate synthesis. However, the emergence of resistant strains has severely compromised the use of pABA mimetics as sulfonamide drugs. Salmonella enterica serovar Gallinarum (S. Gallinarum) is a significant source of antibiotic-resistant infections in poultry. Here, a sulfonamide-resistant FolP mutant library of S. Gallinarum was generated through random mutagenesis. Among resistant strains, substitution of amino acid Arginine 171 with Proline (R171P) in the FolP protein conferred the highest resistance against sulfonamide. Substitution of Phe28 with Leu or Ile (F28L/I) led to modest sulfonamide resistance. Structural modeling indicates that R171P and Phenylalanine 28 with leucine or isoleucine (F28L/I) substitution mutations are located far from the substrate-binding site and cause insignificant conformational changes in the FolP protein. Rather, in silico studies suggest that the mutations altered the stability of the protein, potentially resulting in sulfonamide resistance. Identification of specific mutations in FolP that confer resistance to sulfonamide would contribute to our understanding of the molecular mechanisms of antibiotic resistance.
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Funding
This work was supported by the Starting growth Technological R&D Program (TIPS Program (No. S3130592)) funded by the Ministry of SMEs and Startups (MSS, Korea) in 2021. JSK was supported partly by Chonnam National University (No. 2021–3901).
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TD performed the experiment and analyzed data. J-SK helped to prepare the 3D models. J-HJ and KK provided the helps needed in performing the experiment. HEC wrote the manuscript. All authors read and approved the final manuscript.
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Duysak, T., Jeong, JH., Kim, K. et al. Analysis of random mutations in Salmonella Gallinarum dihydropteroate synthase conferring sulfonamide resistance. Arch Microbiol 205, 363 (2023). https://doi.org/10.1007/s00203-023-03696-5
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DOI: https://doi.org/10.1007/s00203-023-03696-5