A functional metagenomic approach for expanding the synthetic biology toolbox for biomass conversion [electronic resource]

• Ngôn ngữ: eng
• Ký hiệu phân loại: 666.3 Pottery
• Thông tin xuất bản: Oak Ridge, Tenn. : Distributed by the Office of Scientific and Technical Information, U.S. Dept. of Energy, 2010
• Mô tả vật lý: Size: Article No. 360 : , digital, PDF file.
• Bộ sưu tập: Metadata
• ID: 265667
• Chủ đề: 1. 09 biomass fuels; 2. 59 basic biological sciences; 3. Biochemistry & molecular biology; 4. Functional genomics; 5. Synthetic biology

Tóm tắt

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.