Engineering cellulolytic bacterium Clostridium thermocellum to co-ferment cellulose- and hemicellulose-derived sugars simultaneously

Xiong, Wei; Reyes, Luis H.; Michener, William E.; Maness, Pin -Ching; Chou, Katherine J.

doi:10.1002/bit.26590

Title: Engineering cellulolytic bacterium Clostridium thermocellum to co-ferment cellulose- and hemicellulose-derived sugars simultaneously

Journal Article · Tue Apr 10 00:00:00 EDT 2018 · Biotechnology and Bioengineering

DOI:https://doi.org/10.1002/bit.26590· OSTI ID:1430817

Xiong, Wei ^[1]; Reyes, Luis H. ^[2]; Michener, William E. ^[1]; Maness, Pin -Ching ^[1];

^[1]

National Renewable Energy Lab. (NREL), Golden, CO (United States)
National Renewable Energy Lab. (NREL), Golden, CO (United States); Univ. de los Andes, Bogota (Columbia)

Here, cellulose and hemicellulose are the most abundant components in plant biomass. A preferred Consolidated Bioprocessing (CBP) system is one which can directly convert both cellulose and hemicellulose into target products without adding the costly hydrolytic enzyme cocktail. In this work, the thermophilic, cellulolytic, and anaerobic bacterium, Clostridium thermocellum DSM 1313, was engineered to grow on xylose in addition to cellulose. Both xylA (encoding for xylose isomerase) and xylB (encoding for xylulokinase) genes from the thermophilic anaerobic bacterium Thermoanaerobacter ethanolicus were introduced to enable xylose utilization while still retaining its inherent ability to grow on 6-carbon substrates. Targeted integration of xylAB into C. thermocellum genome realized simultaneous fermentation of xylose with glucose, with cellobiose (glucose dimer), and with cellulose, respectively, without carbon catabolite repression. We also showed that the respective H₂ and ethanol production were twice as much when both xylose and cellulose were consumed simultaneously than when consuming cellulose alone. Moreover, the engineered xylose consumer can also utilize xylo-oligomers (with degree of polymerization of 2-7) in the presence of xylose. Isotopic tracer studies also revealed that the engineered xylose catabolism contributed to the production of ethanol from xylan which is a model hemicellulose in mixed sugar fermentation, demonstrating immense potential of this enhanced CBP strain in co-utilizing both cellulose and hemicellulose for the production of fuels and chemicals.

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Research Organization:: National Renewable Energy Laboratory (NREL), Golden, CO (United States)

Sponsoring Organization:: USDOE Office of Energy Efficiency and Renewable Energy (EERE), Hydrogen and Fuel Cell Technologies Program (EE-3F); USDOE Office of Energy Efficiency and Renewable Energy (EERE), NREL Laboratory Directed Research and Development (LDRD); USDOE

Grant/Contract Number:: AC36-08GO28308; 0627-1403

OSTI ID:: 1430817

Alternate ID(s):: OSTI ID: 1432725

Report Number(s):: NREL/JA-2700-71150

Journal Information:: Biotechnology and Bioengineering, Vol. 115, Issue 7; ISSN 0006-3592

Publisher:: WileyCopyright Statement

Country of Publication:: United States

Language:: English

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Cited by: 35 works

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Consolidated bioprocessing for butanol production of cellulolytic Clostridia: development and optimization Wen, Zhiqiang; Li, Qi; Liu, Jinle Microbial Biotechnology, Vol. 13, Issue 2 https://doi.org/10.1111/1751-7915.13478	journal	August 2019

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Title: Engineering cellulolytic bacterium Clostridium thermocellum to co-ferment cellulose- and hemicellulose-derived sugars simultaneously

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