Skip to main content
Log in

Comprehensive optimization of the metabolomic methodology for metabolite profiling of Corynebacterium glutamicum

  • Applied microbial and cell physiology
  • Published:
Applied Microbiology and Biotechnology Aims and scope Submit manuscript

Abstract

Metabolomics has been a potential tool for strain improvement through analyzing metabolite changes in the context of different conditions. However, the availability of a universal metabolite profiling analysis is still a big challenge. In this study, we presented an optimized liquid chromatography-tandem mass spectrometry-based metabolomics methodology for Corynebacterium glutamicum, an important industrial workhorse. It was found that quenching the cellular metabolism with 5-fold volume of − 20 °C 40% methanol was highly recommended due to its lower cell damage rate and higher intracellular metabolite recovery rate. For extracting intracellular metabolites, ethanol/water (3:1, v/v) at 100 °C combined with acidic acetonitrile/water (1:1, v/v, with 0.1% formic acid) at − 20 °C achieved the unbiased metabolite profiling of C. glutamicum. The established methodology was then applied to investigate the intracellular metabolite differences between C. glutamicum ATCC 13032 and an mscCG-deleted mutant under biotin limitation condition. It was observed that in the presence of the functional l-glutamate exporter MscCG, biotin limitation led to accumulation of intracellular 2-oxoglutarate but not l-glutamate. Deletion of mscCG severely inhibited l-glutamate excretion and resulted in a dramatical increase of intracellular l-glutamate, which in turn affected the metabolite profile. The optimized metabolomics methodology holds promise for promoting studies on metabolic mechanism of C. glutamicum.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

Download references

Acknowledgements

This study was supported by the National Natural Sciences Foundation of China (31370113, 31700085, 31700044, and 31370829), Chinese Academy of Sciences Key Project (ZDRW-ZS-2016-2), and the first Special Support Plan for Talents Development and High-Level Innovation and Entrepreneurship Team of the Tianjin Municipal City.

Author information

Authors and Affiliations

Authors

Contributions

P.Z. and J.S. conceived the project. Q.Z. and X.Z. designed the study. Q.Z., J.Y., and Z.Z. carried out the experiments. Q.Z., X.Z., and Y.W. interpreted the results and wrote this manuscript. P.Z., J.S., and Y.M. revised the manuscript and gave valuable suggestion to improve the manuscript. T.D. polished the manuscript. All authors read and approved the final manuscript for publication.

Corresponding authors

Correspondence to Ping Zheng or Jibin Sun.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical statement

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

Electronic supplementary material

ESM 1

(PDF 415 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Zhang, Q., Zheng, X., Wang, Y. et al. Comprehensive optimization of the metabolomic methodology for metabolite profiling of Corynebacterium glutamicum. Appl Microbiol Biotechnol 102, 7113–7121 (2018). https://doi.org/10.1007/s00253-018-9095-1

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00253-018-9095-1

Keywords

Navigation