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The unique difference with the NC case was the synergetic effect manifested between the contact time and temperature (Fig. 10e).
To further probe the relevance of electrostatic interactions manifested between the bacterial cells and the α-HL lumen opening, we conducted experiments at a pH = 4 set in both chambers of the bilayer setup.
This may be also a consequence of the reduced repulsive interactions manifested between the P.a. and the α-HL, which would facilitate the bacterium residence at the α-HL's lumen entrance in acidic pH's as compared to neutral ones.
The charge reduction on bacterial surface leads to an augmented association rate of the P.a.-α-HL interactions, resulting from the reduced magnitude of electrostatic repulsive forces manifested between the negatively charged bacteria and the lumen entrance of the α-HL (Fp-b), which over-compensates for the reduced electrophoretic force (F elp) acting on the bacterium (see text).
We note that among others, the energy barrier term (U*) contains lumped contributions related to electrostatic repulsive interactions manifested between the negatively charged bacterial cells and the lumen entrance of the α-HL nanopore, composed of 14 aspartic acids (D127 and D128) and seven lysines (K131) from the seven protein monomers, which at neutral pH assumes a net negative charge [53].
As we will also demonstrate below, we conclude that electrostatic repulsive interactions manifested between the negatively charged bacteria and the α-HL's lumen entrance contribute to an increase in the free energy barrier needed to be overcome (U*, vide supra), as the bacteria proceeds along its pathway to the nanopore's entrance, leading to its transient association to the α-HL.
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