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Biotechnology Letters

, Volume 40, Issue 2, pp 383–391 | Cite as

Native promoters of Corynebacterium glutamicum and its application in l-lysine production

  • Xiuling Shang
  • Xin Chai
  • Xuemei Lu
  • Yuan Li
  • Yun Zhang
  • Guoqiang Wang
  • Chen Zhang
  • Shuwen Liu
  • Yu Zhang
  • Jiyin Ma
  • Tingyi Wen
Original Research Paper

Abstract

Objective

To identify useful native promoters of Corynebacterium glutamicum for fine-tuning of gene expression in metabolic engineering.

Results

Sixteen native promoters of C. glutamicum were characterized. These promoters covered a strength range of 31-fold with small increments and exhibited relatively stable activity during the whole growth phase using β-galactosidase as the reporter. The mRNA level and enzymatic activity of the lacZ reporter gene exhibited high correlation (R 2 = 0.96) under the control of these promoters. Sequence analysis found that strong promoters had high similarity of the -10 hexamer to the consensus sequence and preference of the AT-rich UP element upstream the -35 region. To test the utility of the promoter library, the characterized native promoters were applied to modulate the sucCD-encoded succinyl-CoA synthetase expression for l-lysine overproduction.

Conclusions

The native promoters with various strengths realize the efficient and precise regulation of gene expression in metabolic engineering of C. glutamicum.

Keywords

Corynebacterium glutamicum Gene expression l-Lysine production Metabolic engineering Native promoters Promoters 

Notes

Acknowledgements

This work was supported by grants from National Hi-Tech Research and Development Program of China (2014AA021203), the Science and Technology Service Network Initiative of the Chinese Academy of Sciences (KFJ-EW-STS-078) and National Natural Science Foundation of China (31100074).

Supporting information

Supplementary Table 1—Strains and plasmids used.

Supplementary Table 2—Primers used.

Supplementary Fig. 1—Sequence analysis of the sixteen promoters.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Research involving human and animal participants

The research performed did not involve human participants and/or animals.

Supplementary material

10529_2017_2479_MOESM1_ESM.docx (389 kb)
Supplementary material 1 (DOCX 389 kb)

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Copyright information

© Springer Science+Business Media B.V., part of Springer Nature 2017

Authors and Affiliations

  1. 1.CAS Key Laboratory of Pathogenic Microbiology and Immunology, Institute of MicrobiologyChinese Academy of SciencesBeijingChina
  2. 2.Beijing Zhongke EPPEN Biotechnology Co., LtdBeijingChina
  3. 3.Ningxia EPPEN Biotechnology Co., LtdYongningChina
  4. 4.Savaid Medical SchoolUniversity of Chinese Academy of SciencesBeijingChina

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