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Adaptive laboratory evolution of ethanologenic Zymomonas mobilis strain tolerant to furfural and acetic acid inhibitors

  • Bioenergy and biofuels
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Abstract

Furfural and acetic acid from lignocellulosic hydrolysates are the prevalent inhibitors to Zymomonas mobilis during cellulosic ethanol production. Developing a strain tolerant to furfural or acetic acid inhibitors is difficul by using rational engineering strategies due to poor understanding of their underlying molecular mechanisms. In this study, strategy of adaptive laboratory evolution (ALE) was used for development of a furfural and acetic acid-tolerant strain. After three round evolution, four evolved mutants (ZMA7-2, ZMA7-3, ZMF3-2, and ZMF3-3) that showed higher growth capacity were successfully obtained via ALE method. Based on the results of profiling of cell growth, glucose utilization, ethanol yield, and activity of key enzymes, two desired strains, ZMA7-2 and ZMF3-3, were achieved, which showed higher tolerance under 7 g/l acetic acid and 3 g/l furfural stress condition. Especially, it is the first report of Z. mobilis strain that could tolerate higher furfural. The best strain, Z. mobilis ZMF3-3, has showed 94.84 % theoretical ethanol yield under 3-g/l furfural stress condition, and the theoretical ethanol yield of ZM4 is only 9.89 %. Our study also demonstrated that ALE method might also be used as a powerful metabolic engineering tool for metabolic engineering in Z. mobilis. Furthermore, the two best strains could be used as novel host for further metabolic engineering in cellulosic ethanol or future biorefinery. Importantly, the two strains may also be used as novel-tolerant model organisms for the genetic mechanism on the “omics” level, which will provide some useful information for inverse metabolic engineering.

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Acknowledgments

This work was supported by Open Funds of State Key Laboratory of Microbial Technology (Shandong University, M2013-07), Open Funds of Key Laboratory of Microbial Resources Collection and Preservation (Ministry of Agriculture, MOA, 2013). Partially supported by Open Funds of Xinjiang Production and Construction Corps Key Laboratory of Protection and Utilization of Biological Resources in Tarim Basin (Tarim University, BRZD1403). Partially supported by Youth Science and Technology Foundation of Sichuan Province (2015JQO047).

Conflict of interests

The authors declare that they have no competing interests.

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Authors and Affiliations

  1. Biogas Institute of Ministry of Agriculture, Biomass Energy Technology Research Centre, Section 4-13, Renming Nanlu, Chengdu, 610041, People’s Republic of China

    Zong-Xia Shui, Han Qin, Bo Wu, Fu-Rong Tan, Jing-Li Wang, Xiao-Yu Tang, Li-Chun Dai, Guo-Quan Hu & Ming-Xiong He

  2. Key Laboratory of Development and Application of Rural Renewable Energy, Ministry of Agriculture, Chengdu, 610041, People’s Republic of China

    Guo-Quan Hu & Ming-Xiong He

  3. Institute of Agricultural Resources and Regional Planning, Chinese Academy of Agricultural Sciences, Beijing, 100081, People’s Republic of China

    Zhi-yong Ruan

  4. State Key Laboratory of Microbial Technology, Shandong University, Jinan, 250100, People’s Republic of China

    Lu-shang Wang

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  1. Zong-Xia Shui
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  2. Han Qin
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Correspondence to Ming-Xiong He.

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Shui, ZX., Qin, H., Wu, B. et al. Adaptive laboratory evolution of ethanologenic Zymomonas mobilis strain tolerant to furfural and acetic acid inhibitors. Appl Microbiol Biotechnol 99, 5739–5748 (2015). https://doi.org/10.1007/s00253-015-6616-z

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  • DOI: https://doi.org/10.1007/s00253-015-6616-z

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