Motility Modulates the Partitioning of Bacteria in Aqueous Two-Phase Systems

We study the partitioning of motile bacteria into different thermodynamic states of a phase-separated dextran (DEX)/polyethylene glycol (PEG) aqueous mixture. While non-motile bacteria partition exclusively into the DEX-rich phase in all conditions tested, we observed that motile bacteria transverse the soft DEX/PEG interface and partition variably among the two phases. For our model organism Bacillus subtilis, the fraction of motile bacteria in the DEX-rich phase increased from 0.58 to 1 as we increased DEX composition within the two-phase region. We hypothesized that the chemical affinity between dextran and the bacteria cell wall acts to weakly confine the bacteria within the DEX-rich phase; however, motility can generate sufficient mechanical forces to overcome the soft confinement and propel the bacteria into the PEG-rich phase. Using optical tweezers to drag a bacterium across the DEX/PEG interface, we developed a model to predict the overall phase partitioning based on a competition between the interfacial forces and bacterial propulsive forces. Our measurements are supported by a theoretical model of dilute active rods embedded within a periodic soft confinement potential.