Metagenomic scaffolds enable combinatorial lignin transformation
| Autor: | Lindsay D. Eltis, Karen Budwill, Steven J. Hallam, David VanInsberghe, Rahul Singh, William W. Mohn, Cameron R. Strachan, Kateryna Ievdokymenko |
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| Rok vydání: | 2014 |
| Předmět: |
Molecular Sequence Data
Lignocellulosic biomass Computational biology macromolecular substances Biology complex mixtures Lignin Metabolic engineering chemistry.chemical_compound Synthetic biology Biorefining Bioprocess Biotransformation Pseudomonas stutzeri Multidisciplinary Base Sequence Escherichia coli K12 business.industry fungi technology industry and agriculture food and beverages Biological Sciences Biotechnology Transformation (genetics) chemistry Metabolic Engineering Transposon mutagenesis Metagenomics business |
| Zdroj: | Proceedings of the National Academy of Sciences of the United States of America. 111(28) |
| ISSN: | 1091-6490 |
| Popis: | Engineering the microbial transformation of lignocellulosic biomass is essential to developing modern biorefining processes that alleviate reliance on petroleum-derived energy and chemicals. Many current bioprocess 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, bioprocess 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 bed bacterial communities 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 rearrayed in nature via horizontal gene transfer. Lignin transformation activity was then demonstrated for one of the predicted gene products encoding a multicopper oxidase to validate the screen. 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. |
| Databáze: | OpenAIRE |
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