Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Understanding a Core Pilin of the Type IVa Pili of Acidithiobacillus thiooxidans, PilV

  • Araceli Hernandez-Sanchez (Geomicrobiologia, Metalurgia, Universidad Autonoma de San Luis Potosi) ;
  • Edgar D. Paez-Perez (Geomicrobiologia, Metalurgia, Universidad Autonoma de San Luis Potosi) ;
  • Elvia Alfaro-Saldana (Geomicrobiologia, Metalurgia, Universidad Autonoma de San Luis Potosi) ;
  • Vanesa Olivares-Illana (Laboratorio de Interacciones Biomoleculares y Cancer. Instituto de Fisica, Universidad Autonoma de San Luis Potosi) ;
  • J. Viridiana Garcia-Meza (Geomicrobiologia, Metalurgia, Universidad Autonoma de San Luis Potosi)
  • Received : 2023.10.25
  • Accepted : 2023.12.29
  • Published : 2024.03.28
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Abstract

Pilins are protein subunits of pili. The pilins of type IV pili (T4P) in pathogenic bacteria are well characterized, but anything is known about the T4P proteins in acidophilic chemolithoautotrophic microorganisms such as the genus Acidithiobacillus. The interest in T4P of A. thiooxidans is because of their possible role in cell recruitment and bacterial aggregation on the surface of minerals during biooxidation of sulfide minerals. In this study we present a successful ad hoc methodology for the heterologous expression and purification of extracellular proteins such as the minor pilin PilV of the T4P of A. thiooxidans, a pilin exposed to extreme conditions of acidity and high oxidation-reduction potentials, and that interact with metal sulfides in an environment rich in dissolved minerals. Once obtained, the model structure of A. thiooxidans PilV revealed the core basic architecture of T4P pilins. Because of the acidophilic condition, we carried out in silico characterization of the protonation status of acidic and basic residues of PilV in order to calculate the ionization state at specific pH values and evaluated their pH stability. Further biophysical characterization was done using UV-visible and fluorescence spectroscopy and the results showed that PilV remains soluble and stable even after exposure to significant changes of pH. PilV has a unique amino acid composition that exhibits acid stability, with significant biotechnology implications such as biooxidation of sulfide minerals. The biophysics profiles of PilV open new paradigms about resilient proteins and stimulate the study of other pilins from extremophiles.

Keywords

Acknowledgement

This work was supported by the National Council of Humanities Science and Technology (CONAHCyT-CB2017-2018 Project A1-S-11505). EDPP acknowledges the support of this research by a postdoctoral scholarship from CONAHCyT (I1200/311/2023). Thanks are expressed to Fernando Rangel for his experimental support and to Alfredo Mendez for suggestions on methodologies for heterologous cloning of PilV. Finally, to Margaret Schroeder for her valuable comments.

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