A functional metagenomic approach for expanding the synthetic biology toolbox for biomass conversion

Sommer, Morten OA; Church, George M.; Dantas, Gautam

doi:10.1038/msb.2010.16

Title: A functional metagenomic approach for expanding the synthetic biology toolbox for biomass conversion

Journal Article · 12 April 2010 · Molecular Systems Biology

DOI: https://doi.org/10.1038/msb.2010.16 · OSTI ID:1623797

Sommer, Morten OA ^[1]; Church, George M. ^[2]; Dantas, Gautam ^[3]

Harvard Medical School, Boston, MA (United States). Dept. of Genetics; DOE/OSTI
Harvard Medical School, Boston, MA (United States). Dept. of Genetics
Washington Univ., St. Louis, MO (United States). School of Medicine, Center for Genome Sciences and Systems Biology, Dept. of Pathology and Immunology

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 cultureindependent 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.

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Research Organization:: Harvard Medical School, Boston, MA (United States)

Sponsoring Organization:: Det Kongelige Danske Videnskabernes Selskab; The Hartmann Foundation; USDOE

OSTI ID:: 1623797

Journal Information:: Molecular Systems Biology, Journal Name: Molecular Systems Biology Journal Issue: 1 Vol. 6; ISSN 1744-4292

Publisher:: WileyCopyright Statement

Country of Publication:: United States

Language:: English

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Land Clearing and the Biofuel Carbon Debt Fargione, J.; Hill, J.; Tilman, D. Science, Vol. 319, Issue 5867 https://doi.org/10.1126/science.1152747	journal	February 2008
Functional Characterization of the Antibiotic Resistance Reservoir in the Human Microflora Sommer, M. O. A.; Dantas, G.; Church, G. M. Science, Vol. 325, Issue 5944 https://doi.org/10.1126/science.1176950	journal	August 2009
Inactivation of antibiotics and the dissemination of resistance genes Davies, J. Science, Vol. 264, Issue 5157 https://doi.org/10.1126/science.8153624	journal	April 1994
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Improvement of Xylose Uptake and Ethanol Production in Recombinant Saccharomyces cerevisiae through an Inverse Metabolic Engineering Approach Jin, Y. -S.; Alper, H.; Yang, Y. -T. Applied and Environmental Microbiology, Vol. 71, Issue 12 https://doi.org/10.1128/AEM.71.12.8249-8256.2005	journal	December 2005
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Novel resistance functions uncovered using functional metagenomic investigations of resistance reservoirs Pehrsson, Erica C.; Forsberg, Kevin J.; Gibson, Molly K. Frontiers in Microbiology, Vol. 4 https://doi.org/10.3389/fmicb.2013.00145	journal	January 2013
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The evolving interface between synthetic biology and functional metagenomics van der Helm, Eric; Genee, Hans J.; Sommer, Morten O. A. Nature Chemical Biology, Vol. 14, Issue 8 https://doi.org/10.1038/s41589-018-0100-x	journal	July 2018
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09 BIOMASS FUELS
59 BASIC BIOLOGICAL SCIENCES
biochemistry & molecular biology
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biomass inhibitor tolerance
functional genomics
functional metagenomics
strain engineering
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Title: A functional metagenomic approach for expanding the synthetic biology toolbox for biomass conversion

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References (52)

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