Metagenomic scaffolds enable combinatorial lignin transformation
Tuesday, April 29, 2014
Exhibit/Poster Hall, lower level (Hilton Clearwater Beach)
Cameron R. Strachan1, David VanInsberghe2, Rahul Singh3, Lindsay D. Eltis4 and Steven J. Hallam4, (1)MetaMixis, Vancouver, BC, Canada, (2)Microbiology, MIT, Cambridge, (3)Department of Microbiology and Immunology, University of British Columbia, Vancouver, BC, Canada, (4)Microbiology and Immunology, University of British Columbia, Vancouver, BC, Canada
Engineering the microbial transformation of lignocellulosic biomass is essential to developing modern biorefining processes that alleviate reliance on petroleum-derived energy and chemical feedstocks. Many current biorefining process streams depend on the genetic tractability of Escherichia coli with a primary emphasis on engineering cellulose/hemicellulose catabolism, small molecule production, and resistance to product inhibition. Conversely, process streams for lignin transformation remain embryonic, with relatively few environmental strains or enzymes implicated. Here we develop a biosensor responsive to monoaromatic lignin transformation products compatible with functional screening in E. coli. We use this biosensor to retrieve metagenomic scaffolds sourced from coal beds conferring an array of lignin transformation phenotypes that synergize in combination. Transposon mutagenesis and comparative sequence analysis of active clones identified genes encoding six functional classes mediating lignin transformation phenotypes that appear to be mobilized in nature via phage- or plasmid-mediated horizontal gene transfer. These results illuminate cellular and community wide networks acting on aromatic polymers and expand the toolkit for engineering recombinant lignin transformation based on ecological design principles.