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

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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.

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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.

Author information

Author notes

  1. Xiuling Shang and Xin Chai have contributed equally to this work.

Authors and Affiliations

  1. CAS Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences, 1 Beichen West Road, Chaoyang District, Beijing, 100101, China

    Xiuling Shang, Yun Zhang, Shuwen Liu, Yu Zhang & Tingyi Wen

  2. Beijing Zhongke EPPEN Biotechnology Co., Ltd, Beijing, 100085, China

    Xin Chai, Xuemei Lu, Yuan Li, Guoqiang Wang, Chen Zhang, Jiyin Ma & Tingyi Wen

  3. Ningxia EPPEN Biotechnology Co., Ltd, Yongning, 750100, Ningxia, China

    Xin Chai, Xuemei Lu, Yuan Li, Guoqiang Wang, Chen Zhang & Jiyin Ma

  4. Savaid Medical School, University of Chinese Academy of Sciences, Beijing, 100049, China

    Tingyi Wen

Authors
  1. Xiuling Shang
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  2. Xin Chai
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  3. Xuemei Lu
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  4. Yuan Li
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  5. Yun Zhang
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  7. Chen Zhang
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  8. Shuwen Liu
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  9. Yu Zhang
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  11. Tingyi Wen
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Corresponding author

Correspondence to Tingyi Wen.

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Research involving human and animal participants

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

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Shang, X., Chai, X., Lu, X. et al. Native promoters of Corynebacterium glutamicum and its application in l-lysine production. Biotechnol Lett 40, 383–391 (2018). https://doi.org/10.1007/s10529-017-2479-y

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  • DOI: https://doi.org/10.1007/s10529-017-2479-y

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