(686b) Syncretic Adaptive Evolution: Combining Multiple Tolerance Phenotypes With Conjugation

Winkler, J. D., Texas A&M University
Kao, K., Texas A&M University

Evolutionary engineering is a widely applicable method to improve strain tolerance and production phenotypes. Typically, after the conclusion of an adaptive laboratory evolution experiment, resistant clones will be isolated and characterized on a genetic, regulatory, and phenotypic level to determine the underlying mechanisms of adaptation. Engineering the discovered tolerance phenotype into another strain then requires extensive manual reconstruction, depending on the number and type of mutations involved. This process must be repeated for every required strain, further slowing the process of strain development. Here, in order to circumvent this laborious process, we present a conjugation-based method to transfer furfural resistance from an evolved donor population to arbitrary recipient strains through simple selection and screening. The feasibility of this technique is demonstrated by endowing previously evolved acetate or n-butanol resistant strains with furfural tolerance, followed by characterization of the multi-tolerant recombinants. Overall, this technique should be a useful new tool for strain engineers to rapidly transfer and examine complex tolerance phenotypes.