Ethanol tolerance in engineered strains of Clostridium thermocellum

Biotechnology for Biofuels and Bioproducts

Research

Yannick J. Bomble¹  Samantha J. Ziegler¹  Shuen Hon²  Nicholas D. Cervenka²  Anthony A. Lanahan²  Daniel G. Olson²  Lee R. Lynd²  Marybeth I. Maloney²  Angel Pech-Canul²  Liang Tian³  Ying Xia⁴ 

[1] Center for Bioenergy Innovation, Oak Ridge National Laboratory, 37830, Oak Ridge, TN, USA;National Renewable Energy Laboratory, 80401, Golden, CO, USA;Thayer School of Engineering at Dartmouth College, 03755, Hanover, NH, USA;Center for Bioenergy Innovation, Oak Ridge National Laboratory, 37830, Oak Ridge, TN, USA;Thayer School of Engineering at Dartmouth College, 03755, Hanover, NH, USA;Center for Bioenergy Innovation, Oak Ridge National Laboratory, 37830, Oak Ridge, TN, USA;Youth Olympic Village, #1-1-602, Jianye District, Nanjing, Jiangsu, China;Thayer School of Engineering at Dartmouth College, 03755, Hanover, NH, USA;College of Chemical and Biological Engineering, Zhejiang University, Hangzhou, China;

关键词: Whole genome sequencing;    Next-generation sequencing;    Tolerance;    Biofuel;    Hungateiclostridium thermocellum;    Ruminiclostridium thermocellum;    Acetivibrio thermocellus;    AdhE;    Alcohol dehydrogenase;    Acetaldehyde dehydrogenase;   

DOI  :  10.1186/s13068-023-02379-z

 received in 2023-07-07, accepted in 2023-08-05,  发布年份 2023

来源: Springer

【 摘 要 】

Clostridium thermocellum is a natively cellulolytic bacterium that is promising candidate for cellulosic biofuel production, and can produce ethanol at high yields (75–80% of theoretical) but the ethanol titers produced thus far are too low for commercial application. In several strains of C. thermocellum engineered for increased ethanol yield, ethanol titer seems to be limited by ethanol tolerance. Previous work to improve ethanol tolerance has focused on the WT organism. In this work, we focused on understanding ethanol tolerance in several engineered strains of C. thermocellum. We observed a tradeoff between ethanol tolerance and production. Adaptation for increased ethanol tolerance decreases ethanol production. Second, we observed a consistent genetic response to ethanol stress involving mutations at the AdhE locus. These mutations typically reduced NADH-linked ADH activity. About half of the ethanol tolerance phenotype could be attributed to the elimination of NADH-linked activity based on a targeted deletion of adhE. Finally, we observed that rich growth medium increases ethanol tolerance, but this effect is eliminated in an adhE deletion strain. Together, these suggest that ethanol inhibits growth and metabolism via a redox-imbalance mechanism. The improved understanding of mechanisms of ethanol tolerance described here lays a foundation for developing strains of C. thermocellum with improved ethanol production.

【 授权许可】

CC BY   
© BioMed Central Ltd., part of Springer Nature 2023

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