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

Abstract 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 sh...

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Main Authors: Morten OA Sommer, George M Church, Gautam Dantas
Format: Article
Language:English
Published: Springer Nature 2010-04-01
Series:Molecular Systems Biology
Subjects:
Online Access:https://doi.org/10.1038/msb.2010.16
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author Morten OA Sommer
George M Church
Gautam Dantas
author_facet Morten OA Sommer
George M Church
Gautam Dantas
author_sort Morten OA Sommer
collection DOAJ
description Abstract 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.
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spelling doaj-art-86c455e2046e474592cf9dcd301005b72025-08-24T12:00:54ZengSpringer NatureMolecular Systems Biology1744-42922010-04-01611710.1038/msb.2010.16A functional metagenomic approach for expanding the synthetic biology toolbox for biomass conversionMorten OA Sommer0George M Church1Gautam Dantas2Department of Genetics, Harvard Medical SchoolDepartment of Genetics, Harvard Medical SchoolDepartment of Pathology and Immunology, Center for Genome Sciences and Systems Biology, Washington University School of MedicineAbstract 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.https://doi.org/10.1038/msb.2010.16biofuelsbiomass inhibitor tolerancefunctional metagenomicsstrain engineeringsynthetic biology
spellingShingle Morten OA Sommer
George M Church
Gautam Dantas
A functional metagenomic approach for expanding the synthetic biology toolbox for biomass conversion
Molecular Systems Biology
biofuels
biomass inhibitor tolerance
functional metagenomics
strain engineering
synthetic biology
title A functional metagenomic approach for expanding the synthetic biology toolbox for biomass conversion
title_full A functional metagenomic approach for expanding the synthetic biology toolbox for biomass conversion
title_fullStr A functional metagenomic approach for expanding the synthetic biology toolbox for biomass conversion
title_full_unstemmed A functional metagenomic approach for expanding the synthetic biology toolbox for biomass conversion
title_short A functional metagenomic approach for expanding the synthetic biology toolbox for biomass conversion
title_sort functional metagenomic approach for expanding the synthetic biology toolbox for biomass conversion
topic biofuels
biomass inhibitor tolerance
functional metagenomics
strain engineering
synthetic biology
url https://doi.org/10.1038/msb.2010.16
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