Applied Microbiology and Biotechnology

, Volume 101, Issue 5, pp 2189–2199 | Cite as

ARTP mutation and genome shuffling of ABE fermentation symbiotic system for improvement of butanol production

  • Chunkai Gu
  • Genyu Wang
  • Shuai Mai
  • Pengfei Wu
  • Jianrong Wu
  • Gehua Wang
  • Hongjuan Liu
  • Jianan Zhang
Bioenergy and biofuels


Butanol is an ideal renewable biofuel which possesses superior fuel properties. Previously, butanol-producing symbiotic system TSH06 was isolated in our lab, with microoxygen tolerance ability. To boost butanol yield for large-scale industrial production, TSH06 was used as parental strain and subjected to atmospheric and room temperature plasma (ARTP) and four rounds of genome shuffling (GS). ARTP mutant and GS strain were co-cultured with facultative anaerobic Bacillus cereus TSH2 to form a symbiotic system with microoxygen tolerance, which was then subjected to fermentation. Relative messenger RNA (mRNA) level of key enzyme gene was measured by real-time PCR. The highest butanol titer of TS4-30 reached 15.63 g/L, which was 34% higher than TSH06 (12.19 g/L). Compared with parental strain, mRNA of acid-forming gene in TS4-30 decreased in acidogenesis phase, while solvent-forming gene increased in solventogenesis phase. This gene expression pattern was consistent with high butanol yield and low acid level in TS4-30. In summary, symbiotic system TS4-30 was obtained with butanol titer improvement and microoxygen tolerance.


Butanol fermentation Symbiotic system TSH06 TS4-30 Genome shuffling Real-time RT-PCR 



Authors’ contributions

GW, GW, SM, and JZ designed the experiments. CG, GW, and SM performed the experiments. CG, GW, and JZ drafted the manuscript. All authors read and approved the final manuscript.

Compliance with ethical standards

This article does not contain any studies with human participants performed by any of the authors.


This work was supported by the Foundation of Key Laboratory for Industrial Biocatalysis (Tsinghua University), Ministry of Education (No. 2015302), National Natural Science Foundation of China (No. 21176141), and National Energy Administration (No. 20131448926). We appreciate Prof. Xinhua Xing and his group for their help in ARTP mutation, which is the basis of this article.

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

This article does not contain any studies with human participants or animals performed by any of the authors.


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Copyright information

© Springer-Verlag Berlin Heidelberg 2017

Authors and Affiliations

  • Chunkai Gu
    • 1
    • 2
  • Genyu Wang
    • 1
  • Shuai Mai
    • 1
  • Pengfei Wu
    • 1
  • Jianrong Wu
    • 2
  • Gehua Wang
    • 1
  • Hongjuan Liu
    • 1
  • Jianan Zhang
    • 1
  1. 1.Institute of Nuclear and New Energy TechnologyTsinghua UniversityBeijingChina
  2. 2.The Key Laboratory of Industrial Biotechnology, Ministry of Education, School of BiotechnologyJiangnan UniversityWuxiChina

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