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Systems Biology-Based Optimization of Extremely Thermophilic Lignocellulose Conversion to Bioproducts

Technical Report ·
DOI:https://doi.org/10.2172/2202361· OSTI ID:2202361
This was a collaborative project involving researchers at the University of Georgia, North Carolina State University, Sanford‐Burnham‐Prebys Med. Discovery Institute and the University of Rhode Island. The over-arching goal was to demonstrate that non-model microorganisms, specifically extreme thermophiles, can be a strategic metabolic engineering platform for industrial biotechnology. We engineered the most thermophilic lignocellulose-degrading organism known, Caldicellulosiruptor bescii (Cbes), which grows optimally near 80°C, and the most thermophilic fermentative organism known, Pyrococcus furiosus (Pfu), which grows optimally at 100°C, to produce several key industrial chemicals. This work leveraged recent breakthrough advances in the development of molecular genetic tools for these organisms, complemented by a deep understanding of its metabolism and physiology gained over the past decade of study in the PIs’ laboratories. We applied the latest metabolic reconstruction and modeling approaches to optimize biomass to product conversion. Bio-processing above 70°C can have important advantages over near-ambient operations. Highly genetically-modified microorganisms usually have a fitness disadvantage and can be easily overtaken in culture when contaminating microbes are present. The high growth temperature of extreme thermophiles precludes growth or survival of virtually any contaminating organism or phage. This reduces operating costs associated with reactor sterilization and maintaining a sterile facility. In addition, at industrial scales, heat production from microbial metabolic activity vastly outweighs heat loss through bioreactor walls such that cooling is required. Extreme thermophiles have the advantage that non-refrigerated cooling water can be used if needed, and heating requirements can be met with low-grade steam typically in excess capacity on plant sites. We assembled a highly interdisciplinary team that brought together all of the expertise for the project to have successful outcomes. This project also built upon and utilized extensive information already available in the PIs’ labs for both Cbes and Pfu to develop models that provide a comprehensive description of these organisms’ physiology and metabolism that were utilized to inform metabolic engineering strategies. The models were validated with experimental data and the results demonstrated that unpretreated lignocellulose has the potential to be converted into value-added industrial chemicals at high loading at bioreactor scale. The data generated from this research were published in twenty-one peer-reviewed papers in international journals with online access.
Research Organization:
Univ. of Georgia, Athens, GA (United States)
Sponsoring Organization:
USDOE Office of Science (SC), Biological and Environmental Research (BER)
DOE Contract Number:
SC0019391
OSTI ID:
2202361
Report Number(s):
DOE-UGA--19391
Country of Publication:
United States
Language:
English

References (20)

Lignocellulose solubilization and conversion by extremely thermophilic Caldicellulosiruptor bescii improves by maintaining metabolic activity journal May 2019
Metabolically engineered Caldicellulosiruptor bescii as a platform for producing acetone and hydrogen from lignocellulose journal August 2020
The biology and biotechnology of the genus Caldicellulosiruptor: recent developments in ‘Caldi World’ journal July 2019
Modification of the glycolytic pathway in Pyrococcus furiosus and the implications for metabolic engineering journal May 2020
Role of cell-substrate association during plant biomass solubilization by the extreme thermophile Caldicellulosiruptor bescii journal February 2023
Engineering the cellulolytic extreme thermophile Caldicellulosiruptor bescii to reduce carboxylic acids to alcohols using plant biomass as the energy source journal August 2020
Plant biomass fermentation by the extreme thermophile Caldicellulosiruptor bescii for co-production of green hydrogen and acetone: Technoeconomic analysis journal March 2022
Fermentative conversion of unpretreated plant biomass: A thermophilic threshold for indigenous microbial growth journal January 2023
Extreme thermophiles as emerging metabolic engineering platforms journal October 2019
Quantitative fermentation of unpretreated transgenic poplar by Caldicellulosiruptor bescii journal August 2019
The thermophilic biomass-degrading bacterium Caldicellulosiruptor bescii utilizes two enzymes to oxidize glyceraldehyde 3-phosphate during glycolysis journal May 2019
Thermophilic microbial deconstruction and conversion of natural and transgenic lignocellulose journal March 2021
Comparative Biochemical and Structural Analysis of Novel Cellulose Binding Proteins (Tāpirins) from Extremely Thermophilic Caldicellulosiruptor Species journal February 2019
Optimizing Strategies for Bio-Based Ethanol Production Using Genome-Scale Metabolic Modeling of the Hyperthermophilic Archaeon, Pyrococcus furiosus journal June 2023
Engineering Caldicellulosiruptor bescii with Surface Layer Homology Domain-Linked Glycoside Hydrolases Improves Plant Biomass Solubilization journal October 2022
Biochemical and Regulatory Analyses of Xylanolytic Regulons in Caldicellulosiruptor bescii Reveal Genus-Wide Features of Hemicellulose Utilization journal November 2022
Transcriptional Regulation of Plant Biomass Degradation and Carbohydrate Utilization Genes in the Extreme Thermophile Caldicellulosiruptor bescii journal June 2021
Genome-Scale Metabolic Model of Caldicellulosiruptor bescii Reveals Optimal Metabolic Engineering Strategies for Bio-based Chemical Production journal June 2021
Use of the lignocellulose-degrading bacterium Caldicellulosiruptor bescii to assess recalcitrance and conversion of wild-type and transgenic poplar journal March 2020
Whither the genus Caldicellulosiruptor and the order Thermoanaerobacterales: phylogeny, taxonomy, ecology, and phenotype journal August 2023

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