Biological functions at high pressure: transcriptome response of Shewanella oneidensis MR-1 to hydrostatic pressure relevant to Titan and other icy ocean worlds.

High hydrostatic pressure (HHP) is a key driver of life's evolution and diversification on Earth. Icy moons such as Titan, Europa, and Enceladus harbor potentially habitable high-pressure environments within their subsurface oceans. Titan, in particular, is modeled to have subsurface ocean pressures ≥ 150 MPa, which are above the highest pressures known to support life on Earth in natural ecosystems. Piezophiles are organisms that grow optimally at pressures higher than atmospheric (0.1 MPa) pressure and have specialized adaptations to the physical constraints of high-pressure environments - up to ~110 MPa at Challenger Deep, the highest pressure deep-sea habitat explored. While non-piezophilic microorganisms have been shown to survive short exposures at Titan relevant pressures, the mechanisms of their survival under such conditions remain largely unelucidated. To better understand these mechanisms, we have conducted a study of gene expression for Shewanella oneidensis MR-1 using a high-pressure experimental culturing system. MR-1 was subjected to short-term (15 min) and long-term (2 h) HHP of 158 MPa, a value consistent with pressures expected near the top of Titan's subsurface ocean. We show that MR-1 is metabolically active in situ at HHP and is capable of viable growth following 2 h exposure to 158 MPa, with minimal pressure training beforehand. We further find that MR-1 regulates 264 genes in response to short-term HHP, the majority of which are upregulated. Adaptations include upregulation of the genes argA, argB, argC, and argF involved in arginine biosynthesis and regulation of genes involved in membrane reconfiguration. MR-1 also utilizes stress response adaptations common to other environmental extremes such as genes encoding for the cold-shock protein CspG and antioxidant defense related genes. This study suggests Titan's ocean pressures may not limit life, as microorganisms could employ adaptations akin to those demonstrated by terrestrial organisms.

Organic Input to Titan's Subsurface Ocean Through Impact Cratering.

Titan has an organic-rich atmosphere and surface with a subsurface liquid water ocean that may represent a habitable environment. In this work, we determined the amount of organic material that can be delivered from Titan's surface to its ocean through impact cratering. We assumed that Titan's craters produce impact melt deposits composed of liquid water that can founder in its lower-density ice crust and estimated the amount of organic molecules that could be incorporated into these melt lenses. We used known yields for HCN and Titan haze hydrolysis to determine the amount of glycine produced in the melt lenses and found a range of possible flux rates of glycine from the surface to the subsurface ocean. These ranged from 0 to 1011 mol/Gyr for HCN hydrolysis and from 0 to 1014 mol/Gyr for haze hydrolysis. These fluxes suggest an upper limit for biomass productivity of ∼103 kgC/year from a glycine fermentation metabolism. This upper limit is significantly less than recent estimates of the hypothetical biomass production supported by Enceladus's subsurface ocean. Unless biologically available compounds can be sourced from Titan's interior, or be delivered from the surface by other mechanisms, our calculations suggest that even the most organic-rich ocean world in the Solar System may not be able to support a large biosphere.