A coculture-coproduction system designed for enhanced carbon conservation through inter-strain CO2 recycling

Abstract

Carbon loss in the form of CO₂ is an intrinsic and persistent challenge faced during conventional and advanced biofuel production from biomass feedstocks. Current mechanisms for increasing carbon conservation typically require the provision of reduced co-substrates as additional reducing equivalents. This need can be circumvented, however, by exploiting the natural heterogeneity of lignocellulosic sugars mixtures and strategically using specific fractions to drive complementary CO₂ emitting vs. CO₂ fixing pathways. As a demonstration of concept, a coculture-coproduction system was developed by pairing two catabolically orthogonal Escherichia coli strains; one converting glucose to ethanol (G2E) and the other xylose to succinate (X2S). ¹³C-labeling studies reveled that G2E + X2S cocultures were capable of recycling 24% of all evolved CO₂ and achieved a carbon conservation efficiency of 77%; significantly higher than the 64% achieved when all sugars are instead converted to just ethanol. In addition to CO₂ exchange, the latent exchange of pyruvate between strains was discovered, along with significant carbon rearrangement within X2S.

Keywords: 13C-fingerprinting; CO2 fixation; Carbon conservation; Coculture.