Applied Microbiology and Biotechnology

, Volume 102, Issue 7, pp 3337–3347 | Cite as

Furfural-tolerant Zymomonas mobilis derived from error-prone PCR-based whole genome shuffling and their tolerant mechanism

  • Suzhen Huang
  • Tingli Xue
  • Zhiquan Wang
  • Yuanyuan Ma
  • Xueting He
  • Jiefang Hong
  • Shaolan Zou
  • Hao Song
  • Minhua Zhang
Applied microbial and cell physiology


Furfural-tolerant strain is essential for the fermentative production of biofuels or chemicals from lignocellulosic biomass. In this study, Zymomonas mobilis CP4 was for the first time subjected to error-prone PCR-based whole genome shuffling, and the resulting mutants F211 and F27 that could tolerate 3 g/L furfural were obtained. The mutant F211 under various furfural stress conditions could rapidly grow when the furfural concentration reduced to 1 g/L. Meanwhile, the two mutants also showed higher tolerance to high concentration of glucose than the control strain CP4. Genome resequencing revealed that the F211 and F27 had 12 and 13 single-nucleotide polymorphisms. The activity assay demonstrated that the activity of NADH-dependent furfural reductase in mutant F211 and CP4 was all increased under furfural stress, and the activity peaked earlier in mutant than in control. Also, furfural level in the culture of F211 was also more rapidly decreased. These indicate that the increase in furfural tolerance of the mutants may be resulted from the enhanced NADH-dependent furfural reductase activity during early log phase, which could lead to an accelerated furfural detoxification process in mutants. In all, we obtained Z. mobilis mutants with enhanced furfural and high concentration of glucose tolerance, and provided valuable clues for the mechanism of furfural tolerance and strain development.


Furfural tolerance Zymomonas mobilis Furfural reductase Error-prone PCR-based whole genome shuffling Genome resequencing 



We thank Dr. Jie Bao for providing plasmid pHW20a.


This research was supported by the National Natural Science Foundation of China (NSFC30900033, NSFC21376174) and the Tianjin Science and Technology Council (13JCYBJC40700).

Compliance with ethical standards

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.

Supplementary material

253_2018_8817_MOESM1_ESM.pdf (1 mb)
ESM 1 (PDF 1048 kb)


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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  • Suzhen Huang
    • 1
    • 2
  • Tingli Xue
    • 1
    • 2
  • Zhiquan Wang
    • 1
    • 2
  • Yuanyuan Ma
    • 1
    • 3
  • Xueting He
    • 1
    • 2
  • Jiefang Hong
    • 1
  • Shaolan Zou
    • 1
  • Hao Song
    • 3
    • 4
  • Minhua Zhang
    • 1
    • 3
  1. 1.R&D Center for Petrochemical TechnologyTianjin UniversityTianjinChina
  2. 2.Department of Biochemical Engineering, School of Chemical Engineering and TechnologyTianjin UniversityTianjinChina
  3. 3.Collaborative Innovation Centre of Chemical Science and Engineering (Tianjin), School of Chemical Engineering and TechnologyTianjinChina
  4. 4.Key Laboratory of Systems Bioengineering (Ministry of Education), School of Chemical Engineering and TechnologyTianjin UniversityTianjinChina

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