Acidithiobacillus spp. have traditionally been utilized to extract metals from mineral ores through bioleaching. This process has recently expanded to include artificial ores, such as those derived from municipal solid waste incineration (MSWI) residues. Previous studies have indicated that microbial adaptation enhances bioleaching efficiency, prompting this study to identify proteins involved in the adaptation of A. ferridurans to MSWI residues. We employed data-independent acquisition-parallel accumulation serial fragmentation to determine the proteomic response of A. ferridurans DSM 583 to three distinct materials: bottom ash (BA), kettle ash (KA), and filter ash (FA), which represent typical MSWI residues. Our findings indicate that, irrespective of the residue type, a suite of membrane transporters, porins, efflux pumps, and specific electron and cation transfer proteins was notably upregulated. The upregulation of certain proteins involved in anaerobic pathways suggested the development of a spontaneous microaerobic environment, which minimally impacted the bioleaching efficiency. Additionally, the adaptation was most efficient at half the target FA concentration, marked by a significant increase in the detoxification and efflux systems required by microorganisms to tolerate high heavy metal concentrations. Given that metal recovery peaked at lower FA concentrations for most metals of interest, further adaptation at the level of protein expression may not be warranted for improved bioleaching outcomes.

Department of Biochemistry Faculty of Science Masaryk University Brno 625 00 Czech Republic

Department of Chemistry Faculty of Science Masaryk University Brno 625 00 Czech Republic

Department of Geological Sciences Faculty of Science Masaryk University Brno 611 37 Czech Republic

Proteomics Core Facility Central European Institute for Technology Masaryk University Brno 625 00 Czech Republic

University of Natural Resources and Life Sciences Vienna BOKU Department of Agrobiotechnology IFA Tulln Institute of Environmental Biotechnology Tulln and der Donau 3430 Austria

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