26 Sep 2023

Isolation and Characterization of Bacterial Isolates from Alaskan Permafrost for Synthetic Biology Applications

Abstract: Operations in the Artic and other cold regions require technologies that can perform reliably under extreme cold conditions. Permafrost and frozen soils harbor a wide range of microorganisms that have adapted to extremely low temperatures and have unique metabolic capabilities relevant to military operations and that could be exploited to develop biotechnologies optimized for cold environments. Cold-tolerant bacteria (psychrophiles and psychrotrophs) are critical to the development of synthetic biology technologies meant to work in cold environments like the Arctic. Using bacteria isolated from Alaskan permafrost, we applied an experimental pipeline to test the best candidates for use as biological platforms, or chassis, for low-temperature synthetic biology. Since synthetic biology constructs will perform only as well as their chassis, it is critical that circuits expected to perform under extreme cold conditions are housed in chassis that are adapted to those conditions. We identified one permafrost isolate, PTI8, related to Rhodococcus fascians, that is capable of growing from −1°C to at least 25°C and which we experimentally confirmed to uptake and express the broad host range plasmid pBTK519, suggesting PTI8 is a candidate for use as a novel cold-adapted chassis for synthetic biology.