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    Solution and interfacial self-assembly of Bacillus subtilis bacterial lipoteichoic acid (LTA): nanoclustering, and effects of Ca2+ and temperature

    Year: 2022

    Journal: Nanoscale, Volume 14, SEP 2, page 12265–12274

    Authors: Bharatiya, Bhavesh; Wlodek, Magdalena; Harniman, Robert; Schweins, Ralf; Mantell, Judith; Wang, Gang; Warszynski, Piotr; Briscoe, Wuge H.

    Organizations: European Union [797038, 793574]; Engineering and Physical Science Research Council (EPSRC) [EP/H034862/1, EP/H014861/1]; Ministerstwo Edukacji i Nauki via the statutory subsidy for Jerzy Haber Institute of Catalysis and Surface Chemistry PAS

    Lipoteichoic acid (LTA) is a major structural and functional molecule in the Gram-positive bacteria membrane. Knowledge of LTA adsorption at interfaces and its solution self-assembly is crucial to understanding its role in bacterial adhesion and colonisation, infections and inflammations. Here, we report the self-assembly behaviour of LTA extracted from Bacillus subtilis, a Gram-positive bacterium, in an aqueous solution using cryogenic transmission electron microscopy (Cryo-TEM) and small-angle neutron scattering (SANS) and its adsorption behaviour at the solid-liquid interface using atomic force microscopy (AFM) imaging and quartz crystal microbalance with dissipation monitoring (QCM-D). The Cryo-TEM results indicated the formation of spherical LTA micelles that decreased in size on addition of calcium chloride (CaCl2), attributed to charge neutralisation and possible formation of stable Ca2+-bridges between the phosphate groups on neighbouring LTA chains. Analysis of the SANS data from the polydisperse LTA aggregates in solution using the two Lorentzian model revealed the existence of two correlation lengths, which could respectively account for the presence of LTA micelle clusters and the local structure arising from LTA intra-molecular interactions. In the presence of CaCl2, the decrease in the correlation lengths of the clusters indicated possible disruption of H-bonding by Ca2+, leading to poorer water-LTA interactions. At higher temperatures, the correlation length corresponding to the clusters increased, indicating a temperature assisted growth caused by the fluidization of micellar core and dehydration of the polar LTA chains. AFM imaging showed that adsorption of LTA aggregates at the SiO2-water interface was significantly prompted by the addition of CaCl2, also confirmed by QCM-D measurements. These unprecedented nanoscopic structural details on the morphology of LTA aggregates in solution and at the solid-liquid interface add to our fundamental understanding of its self-assembly behaviour hitherto underexplored.
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