Open AccessA functional metagenomic approach for expanding the synthetic biology toolbox for biomass conversion
Author(s) -
Sommer Morten OA,
Church George M,
Dantas Gautam
Publication year - 2010
Publication title -
molecular systems biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 8.523
H-Index - 148
ISSN - 1744-4292
DOI - 10.1038/msb.2010.16
Subject(s) - metagenomics , fosmid , biology , computational biology , biomass (ecology) , synthetic biology , microbiology and biotechnology , gene , biofuel , biochemical engineering , genetics , ecology , engineering
Sustainable biofuel alternatives to fossil fuel energy are hampered by recalcitrance and toxicity of biomass substrates to microbial biocatalysts. To address this issue, we present a culture‐independent functional metagenomic platform for mining Nature's vast enzymatic reservoir and show its relevance to biomass conversion. We performed functional selections on 4.7 Gb of metagenomic fosmid libraries and show that genetic elements conferring tolerance toward seven important biomass inhibitors can be identified. We select two metagenomic fosmids that improve the growth of Escherichia coli by 5.7‐ and 6.9‐fold in the presence of inhibitory concentrations of syringaldehyde and 2‐furoic acid, respectively, and identify the individual genes responsible for these tolerance phenotypes. Finally, we combine the individual genes to create a three‐gene construct that confers tolerance to mixtures of these important biomass inhibitors. This platform presents a route for expanding the repertoire of genetic elements available to synthetic biology and provides a starting point for efforts to engineer robust strains for biofuel generation.