Exometabolomics Assisted Design and Validation of Synthetic Obligate Mutualism

ACS Synthetic Biology
Suzanne M KosinaTrent R Northen

Abstract

Synthetic microbial ecology has the potential to enhance the productivity and resiliency of biotechnology processes compared to approaches using single isolates. Engineering microbial consortia is challenging; however, one approach that has attracted significant attention is the creation of synthetic obligate mutualism using auxotrophic mutants that depend on each other for exchange or cross-feeding of metabolites. Here, we describe the integration of mutant library fitness profiling with mass spectrometry based exometabolomics as a method for constructing synthetic mutualism based on cross-feeding. Two industrially important species lacking known ecological interactions, Zymomonas mobilis and Escherichia coli, were selected as the test species. Amino acid exometabolites identified in the spent medium of Z. mobilis were used to select three corresponding E. coli auxotrophs (proA, pheA and IlvA), as potential E. coli counterparts for the coculture. A pooled mutant fitness assay with a Z. mobilis transposon mutant library was used to identify mutants with improved growth in the presence of E. coli. An auxotroph mutant in a gene (ZMO0748) with sequence similarity to cysteine synthase A (cysK), was selected as the Z. mobilis counte...Continue Reading

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Citations

Oct 21, 2016·Microbial Biotechnology·Shihui YangMin Zhang
Oct 5, 2017·Metabolites·Andrea LubbeTrent Northen
Apr 6, 2018·Nature Biotechnology·Carlos GuijasGary Siuzdak
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