Distinct Acetate Utilization Strategies Differentiate Butyrate and Medium-Chain Carboxylic Acid Producing Chain-Elongating Bacteria

This is the authors' abstract. We don't add key points for this paper.

Chain elongating bacteria (CEB) are a unique guild of anaerobes that upcycle organic waste into valuable short- and medium-chain carboxylic acids (MCCAs), enabling a circular bioeconomy. However, the metabolic rules that determine product chain length have remained elusive. Here, we combine 13 C isotope tracing, proteomics, enzyme assays, and metabolic modelling to show that distinct acetate utilization strategies underlie the divergence between butyrate- and MCCA-producing CEB. MCCA-producing strains recycle acetate to maximize lactate use under acetate limitation, but at the cost of slower growth. In contrast, butyrate-producing strains grow faster by favoring acetate assimilation, at the cost of restricted lactate utilization when acetate is scarce. These physiological trade-offs are encoded in the substrate specificity of coenzyme A transferase, the terminal enzyme in reverse β-oxidation. Our findings uncover a fundamental constraint shaping chain-length selectivity in CEB and offer new strategies to optimize MCCA production from organic waste streams.