Surface reactivity of the iron and manganese-oxidizing bacterium Leptothrix cholodnii SP-6
Authors: Araujo Maicon , Gutierrez Rueda Daniela , Li Yuhao , Kenney Janice , Alessi Daniel , Konhauser Kurt 
Published: October 2023

Surfaces of prokaryotic cells play a significant role in the adsorption of metals from aqueous solution and the formation of authigenic minerals (Konhauser 2006). Although most studies focus on the cell wall, it is known that many bacteria synthesise an extracellular layer of polysaccharides and proteins, including what are known as sheaths. It has been shown that the cyanobacterium Calothrix sp. produces as sheath which is neutrally charged at circumneutral pH values, and it was hypothesized that such a sheath might allow the cyanobacterium to survive in geothermal settings with high silicification rates (Phoenix et al. 2002). Specifically, the dominance of hydroxyl sites on Calothrix’s sheath surface facilitates hydrogen bonding with aqueous silica species, inducing the precipitation of amorphous silica on the sheath and thus protecting the underlying cell (Phoenix et al. 2002). Leptothrix cholodnii is a sheathed, iron and manganese-oxidizing bacterium that frequently inhabits minerals seeps, where Fe2+ and Mn2+ discharge into oxygenated surface waters (Spring et al. 1996). As a result, the sheath becomes encrusted with Fe(III) and Mn(IV) oxyhydroxides while the underlying cells are protected from mineralization (Emerson and Ghiorse 1992, Emerson et al. 2010). However, unlike Calothrix, Leptothrix’s sheath composition suggests that it might behave differently at circumneutral pH (Emerson and Ghiorse 1993). To investigate the surface reactivity of Leptothrix's sheath and cell wall we analyzed isolated sheaths, sheathless cells, and intact filaments of L. cholodnii SP-6. We studied these components using potentiometric titration, zeta-potential, Cd-adsorption, and Fourier transform infrared (FTIR) spectroscopy to elucidate changes in surface charge between the cell wall and sheath. For the isolated sheaths and intact filaments, titration data were fit using a two-site protonation model, resulting in the following pKa values: 6.05 (±0.29) and 9.34 (±0.11); and 7.77 (±0.17) and 10.50 (±0.20), respectively. For the sheathless cells, the best fit was obtained by using a three-site protonation model, resulting in the following pKa values: 5.40 (±0.59), 8.11 (±1.64) and 10.73 (±0.45). Total proton-active site concentrations were lower in isolated sheaths compared to intact filaments. Additionally, at circumneutral pH, net negative charge was lower for sheathless cells compared to intact filaments and isolated sheaths (Fig. 1). This information agrees with the Cd adsorption behaviour found for the three materials (Fig. 2). Thus, our preliminary results suggest that Leptothrix’s sheath is less reactive than the intact filaments at circumneutral pH, leading us to hypothesize that the outermost layer would sequester relatively lower amounts of cations, including Mn2+, from solution and potentially would protect the underlaying cell from deleterious mineralization. In addition to that, the less reactive sheath’s surface would also contribute to cell attachment, which is important for a species commonly found in streams (Phoenix et al. 2002, Emerson et al. 2010).