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Abstract Methane clathrates on continental margins contain the largest stores of hydrocarbons on Earth, yet the role of biomolecules in clathrate formation and stability remains almost completely unknown. Here, we report new methane clathrate-binding proteins (CbpAs) of bacterial origin discovered in metagenomes from gas clathrate-bearing ocean sediments. CbpAs show similar suppression of methane clathrate growth as the commercial gas clathrate inhibitor polyvinylpyrrolidone and inhibit clathrate growth at lower concentrations than antifreeze proteins (AFPs) previously tested. Unlike AFPs, CbpAs are selective for clathrate over ice. CbpA3 adopts a nonglobular, extended structure with an exposed hydrophobic surface, and, unexpectedly, its TxxxAxxxAxx motif common to AFPs is buried and not involved in clathrate binding. Instead, simulations and mutagenesis suggest a bipartite interaction of CbpAs with methane clathrate, with the pyrrolidine ring of a highly conserved proline residue mediating binding by filling empty clathrate cages. The discovery that CbpAs exert such potent control on methane clathrate properties implies that biomolecules from native sediment bacteria may be important for clathrate stability and habitability. 

Immiscible multiphase flow in porous media is largely affected by interfacial properties, manifested in contact angle and surface tension. The gas-liquid surface tension can be significantly altered by suspended particles at the interface. Particle-laden interfaces have unique properties, for example, a lower surface tension of interfaces laden with surfactants or nanoparticles. This study investigates the impacts of a motile bacterium Escherichia coli ( E. coli , strain ATCC 9637) on the air-water surface tension. Methods of the maximum bubble pressure, the du Noüy ring, and the pendant droplet are used to measure the surface tension of the motile-bacteria-laden interfaces. Measured surface tension remains independent to the E. coli concentration when using the maximum bubble pressure method, decreases with increased E. coli concentration in the du Noüy ring method, and presents time-dependent changes by the pendant drop method. The analyses show that the discrepancies may come from the different convection-diffusion processes of E. coli in the flow among various testing methods. 

Multiphase flow patterns in porous media largely depend on the properties of the fluids and interfaces such as viscosity, surface tension, and contact angle. Microorganisms in soils change the fluid and interfacial properties, and thus can alter multiphase fluid flow in porous media. This study investigates the impact of motile bacterium Escherichia coli ( E. coli ) on fluid displacement patterns in a microfluidic chip. The fluid displacement is observed during the saturation and the desaturation processes of the microfluidic chip with and without E.coli suspension. Time-lapse photography results show that the presence of E.coli alters the displacement patterns during the wetting and drying process and changes the residual saturation of the chip. Although studies of the impacts of motility on interfacial properties remain elusive, these results bring the expectation to the manipulation of multiphase transport in porous media and the adaptive control of industrial and environmental flow processes using active particles.