skip to main content
OSTI.GOV title logo U.S. Department of Energy
Office of Scientific and Technical Information

Title: Multi-omic elucidation of aromatic catabolism in adaptively evolved Rhodococcus opacus

Journal Article · · Metabolic Engineering

Lignin utilization has been identified as a key factor in biorefinery profitability. However, lignin depolymerization generates heterogeneous aromatic mixtures that inhibit microbial growth and conversion of lignocellulose to biochemicals. Rhodococcus opacus is a promising aromatic-catabolizing, oleaginous bacterium, but mechanisms for its aromatic tolerance and utilization remain undercharacterized. To better understand these mechanisms, we adaptively evolved R. opacus for improved utilization of 32 combinations of diverse aromatic compounds. Evolved R. opacus mutants showed up to 1900% growth improvement in the utilization of phenol, guaiacol, 4-hydroxybenzoate, vanillate, and benzoate compared to the wild-type strain. Whole genome sequencing revealed several redox-related genes with mutations shared across multiple adapted mutants. PVHG6, the mutant with the most improved growth on a mixture of multiple aromatic compounds, showed 56% lower superoxide dismutase activity than the wild-type strain, suggesting that redox reactions are important for aromatic tolerance and utilization. Comparative transcriptomics revealed by-product detoxification pathways and five aromatic funneling pathways that were upregulated in response to specific aromatic compounds. Gene knockout experiments confirmed the two degradation routes of the β-ketoadipate pathway for five aromatic compounds. Finally, these results provide an improved understanding of aromatic bioconversion and facilitate development of R. opacus as a biorefinery host.

Research Organization:
Washington Univ., St. Louis, MO (United States)
Sponsoring Organization:
USDOE Office of Science (SC), Biological and Environmental Research (BER)
Contributing Organization:
Gautam Dantas
Grant/Contract Number:
SC0012705
OSTI ID:
1544572
Alternate ID(s):
OSTI ID: 1548002
Journal Information:
Metabolic Engineering, Vol. 49, Issue C; ISSN 1096-7176
Publisher:
ElsevierCopyright Statement
Country of Publication:
United States
Language:
English
Citation Metrics:
Cited by: 37 works
Citation information provided by
Web of Science

References (85)

Formation of intracytoplasmic lipid inclusions by Rhodococcus opacus strain PD630 journal June 1996
Gene Ontology: tool for the unification of biology journal May 2000
Review of US and EU initiatives toward development, demonstration, and commercialization of lignocellulosic biofuels journal August 2013
Inexpensive Multiplexed Library Preparation for Megabase-Sized Genomes journal May 2015
Harnessing Yarrowia lipolytica lipogenesis to create a platform for lipid and biofuel production journal January 2014
Trimmomatic: a flexible trimmer for Illumina sequence data journal April 2014
Oxygenases: mechanisms and structural motifs for O2 activation journal October 2001
Enzymatic conversion of lignin into renewable chemicals journal December 2015
BLAST+: architecture and applications journal January 2009
Vanillin Catabolism in Rhodococcus jostii RHA1 journal November 2011
Integrated omics study delineates the dynamics of lipid droplets in Rhodococcus opacus PD630 journal October 2013
Benzoate Catabolite Repression of the Phthalate Degradation Pathway in Rhodococcus sp. Strain DK17 journal December 2006
A Mobile Element in mutS Drives Hypermutation in a Marine Vibrio journal February 2017
Microbial laboratory evolution in the era of genome‐scale science journal January 2011
The demethylation of guaiacol by a new bacterial cytochrome P-450 journal February 1985
Development of Chemical and Metabolite Sensors for Rhodococcus opacus PD630 journal June 2017
Molecular Toolkit for Gene Expression Control and Genome Modification in Rhodococcus opacus PD630 journal January 2018
A framework for variation discovery and genotyping using next-generation DNA sequencing data journal April 2011
The evolution of a conjugative plasmid and its ability to increase bacterial fitness journal May 2005
Engineering microbes for tolerance to next-generation biofuels journal January 2011
Selection and optimization of microbial hosts for biofuels production journal November 2008
A Simple Method for the Isolation and Purification of Total Lipides from Animal Tissues journal May 1957
The mobile flavin of 4-OH benzoate hydroxylase journal October 1994
High-throughput functional annotation and data mining with the Blast2GO suite journal April 2008
Gene redundancy of two-component (chloro)phenol hydroxylases in Rhodococcus opacus 1CP journal October 2014
Bio-ethanol – the fuel of tomorrow from the residues of today journal December 2006
Plasmid-mediated horizontal gene transfer is a coevolutionary process journal June 2012
The β-Ketoadipate Pathway And The Biology Of Self-Identity journal October 1996
Transcriptomic characterization of Caecomyces churrovis: a novel, non-rhizoid-forming lignocellulolytic anaerobic fungus journal December 2017
Comparative and Functional Genomics of Rhodococcus opacus PD630 for Biofuels Development journal September 2011
Molecular Adaptation Mechanisms Employed by Ethanologenic Bacteria in Response to Lignocellulose-derived Inhibitory Compounds journal January 2013
Metabolic engineering of yeast for lignocellulosic biofuel production journal December 2017
Experimental and computational optimization of an Escherichia coli co-culture for the efficient production of flavonoids journal May 2016
InterProScan 5: genome-scale protein function classification journal January 2014
Two independently regulated cytochromes P-450 in a Rhodococcus rhodochrous strain that degrades 2-ethoxyphenol and 4-methoxybenzoate. journal March 1993
Aromatic inhibitors derived from ammonia-pretreated lignocellulose hinder bacterial ethanologenesis by activating regulatory circuits controlling inhibitor efflux and detoxification journal August 2014
The Phyre2 web portal for protein modeling, prediction and analysis journal May 2015
A "Silent" Polymorphism in the MDR1 Gene Changes Substrate Specificity journal January 2007
The cytochrome bcc-aa3-type respiratory chain of Rhodococcus rhodochrous journal July 2010
Bioconversion of lignin model compounds with oleaginous Rhodococci journal December 2011
High-cell-density batch fermentation of Rhodococcus opacus PD630 using a high glucose concentration for triacylglycerol production journal June 2010
Tolerance and adaptive evolution of triacylglycerol-producing Rhodococcus opacus to lignocellulose-derived inhibitors journal January 2015
Engineering xylose metabolism in triacylglycerol-producing Rhodococcus opacus for lignocellulosic fuel production journal January 2013
Fast gapped-read alignment with Bowtie 2 journal March 2012
Utilization of simultaneous saccharification and fermentation residues as feedstock for lipid accumulation in Rhodococcus opacus journal September 2017
Cumulative Number of Cell Divisions as a Meaningful Timescale for Adaptive Laboratory Evolution of Escherichia coli journal October 2011
Homo-succinic acid production by metabolically engineered Mannheimia succiniciproducens journal November 2016
Dynamics of adaptation and diversification: a 10,000-generation experiment with bacterial populations. journal July 1994
Formaldehyde degradation in Corynebacterium glutamicum involves acetaldehyde dehydrogenase and mycothiol-dependent formaldehyde dehydrogenase journal December 2013
The Sequence Alignment/Map format and SAMtools journal June 2009
featureCounts: an efficient general purpose program for assigning sequence reads to genomic features journal November 2013
Lignin valorization through integrated biological funneling and chemical catalysis journal August 2014
Moderated estimation of fold change and dispersion for RNA-seq data with DESeq2 journal December 2014
Using design of experiments to optimize derivatization with methyl chloroformate for quantitative analysis of the aqueous phase from hydrothermal liquefaction of biomass journal January 2016
Superoxide dismutases: Ancient enzymes and new insights journal November 2011
Assembling Single-Cell Genomes and Mini-Metagenomes From Chimeric MDA Products journal October 2013
The Ribulose Monophosphate Pathway Substitutes for the Missing Pentose Phosphate Pathway in the Archaeon Thermococcus kodakaraensis journal June 2006
Lignin Depolymerization and Conversion A Review of Thermochemical Methods journal November 2010
Proteomic Analysis of Survival of Rhodococcus jostii RHA1 during Carbon Starvation journal July 2012
Efficient utilization of pentoses for bioproduction of the renewable two-carbon compounds ethylene glycol and glycolate journal March 2016
Engineering a riboswitch-based genetic platform for the self-directed evolution of acid-tolerant phenotypes journal September 2017
Role of pretreatment and conditioning processes on toxicity of lignocellulosic biomass hydrolysates journal June 2009
Molecular characterization of genes of Pseudomonas sp. strain HR199 involved in bioconversion of vanillin to protocatechuate. journal April 1997
Lipid production in Yarrowia lipolytica is maximized by engineering cytosolic redox metabolism journal January 2017
Lignin Valorization: Improving Lignin Processing in the Biorefinery journal May 2014
Base-Catalyzed Depolymerization of Solid Lignin-Rich Streams Enables Microbial Conversion journal August 2017
Evolution of Escherichia coli to 42 °C and Subsequent Genetic Engineering Reveals Adaptive Mechanisms and Novel Mutations journal July 2014
Formaldehyde stabilization facilitates lignin monomer production during biomass depolymerization journal October 2016
Evolution of high mutation rates in experimental populations of E. coli journal June 1997
Benzoate degradation by Rhodococcus opacus 1CP after dormancy: Characterization of dioxygenases involved in the process journal December 2015
Hydrolysis of lignocellulosic materials for ethanol production: a review journal May 2002
Effect of Aromatic Compounds on Cellular Fatty Acid Composition of Rhodococcus opacus journal January 1999
Biofuels 2020: Biorefineries based on lignocellulosic materials journal July 2016
Integrating lignin valorization and bio-ethanol production: on the role of Ni-Al 2 O 3 catalyst pellets during lignin-first fractionation journal January 2017
Adipic acid production from lignin journal January 2015
Pilon: An Integrated Tool for Comprehensive Microbial Variant Detection and Genome Assembly Improvement journal November 2014
Integration of heterogeneous and biochemical catalysis for production of fuels and chemicals from biomass journal June 2017
Advanced Chemical Design for Efficient Lignin Bioconversion journal January 2017
Engineering Yarrowia lipolytica as a platform for synthesis of drop-in transportation fuels and oleochemicals journal September 2016
One-step fermentative production of aromatic polyesters from glucose by metabolically engineered Escherichia coli strains journal January 2018
SuSPect: Enhanced Prediction of Single Amino Acid Variant (SAV) Phenotype Using Network Features journal July 2014
Comparative transcriptomics elucidates adaptive phenol tolerance and utilization in lipid-accumulating Rhodococcus opacus PD630 journal February 2016
Gene Expression Analysis of Methylotrophic Oxidoreductases Involved in the Oligotrophic Growth of Rhodococcus erythropolis N9T-4 journal January 2011
Harnessing Yeast Peroxisomes for Biosynthesis of Fatty-Acid-Derived Biofuels and Chemicals with Relieved Side-Pathway Competition journal October 2016
Production of fatty acid-derived oleochemicals and biofuels by synthetic yeast cell factories journal May 2016

Cited By (3)

Potential of Bacillus subtilis from marine environment to degrade aromatic hydrocarbons journal July 2019
Mapping the diversity of microbial lignin catabolism: experiences from the eLignin database journal April 2019
Development of Rhodococcus opacus as a chassis for lignin valorization and bioproduction of high-value compounds journal August 2019