Dataset Information

Optimization of a heterologous pathway for protocatechuate catabolism in Escherichia coli

ABSTRACT: Optimization of a heterologous pathway for protocatechuate catabolism in Escherichia coli

INSTRUMENT(S): Q Exactive Plus

ORGANISM(S): Escherichia Coli (ncbitaxon:562)

SUBMITTER: Joshua K. Michener

PROVIDER: MSV000081122 | MassIVE | Tue May 30 13:48:00 BST 2017

SECONDARY ACCESSION(S): PXD006627

REPOSITORIES: MassIVE

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Publications

Construction and Optimization of a Heterologous Pathway for Protocatechuate Catabolism in Escherichia coli Enables Bioconversion of Model Aromatic Compounds.

Clarkson Sonya M SM Giannone Richard J RJ Kridelbaugh Donna M DM Elkins James G JG Guss Adam M AM Michener Joshua K JK

Applied and environmental microbiology 20170831 18

The production of biofuels from lignocellulose yields a substantial lignin by-product stream that currently has few applications. Biological conversion of lignin-derived compounds into chemicals and fuels has the potential to improve the economics of lignocellulose-derived biofuels, but few microbes are able both to catabolize lignin-derived aromatic compounds and to generate valuable products. While <i>Escherichia coli</i> has been engineered to produce a variety of fuels and chemicals, it is i ...[more]

PMID: 28733280

Similar Datasets

Project description:The production of biofuels from lignocellulose yields a substantial lignin by-product stream that currently has few applications. Biological conversion of lignin-derived compounds into chemicals and fuels has the potential to improve the economics of lignocellulose-derived biofuels, but few microbes are able both to catabolize lignin-derived aromatic compounds and to generate valuable products. While Escherichia coli has been engineered to produce a variety of fuels and chemicals, it is incapable of catabolizing most aromatic compounds. Therefore, we engineered E. coli to catabolize protocatechuate, a common intermediate in lignin degradation, as the sole source of carbon and energy via heterologous expression of a nine-gene pathway from Pseudomonas putida KT2440. We next used experimental evolution to select for mutations that increased growth with protocatechuate more than 2-fold. Increasing the strength of a single ribosome binding site in the heterologous pathway was sufficient to recapitulate the increased growth. After optimization of the core pathway, we extended the pathway to enable catabolism of a second model compound, 4-hydroxybenzoate. These engineered strains will be useful platforms to discover, characterize, and optimize pathways for conversions of lignin-derived aromatics.IMPORTANCE Lignin is a challenging substrate for microbial catabolism due to its polymeric and heterogeneous chemical structure. Therefore, engineering microbes for improved catabolism of lignin-derived aromatic compounds will require the assembly of an entire network of catabolic reactions, including pathways from genetically intractable strains. Constructing defined pathways for aromatic compound degradation in a model host would allow rapid identification, characterization, and optimization of novel pathways. We constructed and optimized one such pathway in E. coli to enable catabolism of a model aromatic compound, protocatechuate, and then extended the pathway to a related compound, 4-hydroxybenzoate. This optimized strain can now be used as the basis for the characterization of novel pathways.