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Biotechnology Letters

, Volume 41, Issue 1, pp 69–77 | Cite as

Deletion of gene gnd encoding 6-phosphogluconate dehydrogenase promotes l-serine biosynthesis in a genetically engineered strain of Methylobacterium sp. MB200

  • Xian Li
  • Bo Wu
  • Kan Zhou
  • Chengjian Jiang
  • Peihong Shen
Original Research Paper
  • 126 Downloads

Abstract

Objective

To identify potential target genes involved in l-serine biosynthesis in Methylobacterium sp. MB200 and to evaluate the gnd genetically-engineered strains for l-serine production.

Results

Five genes that are not associated with the central metabolic pathway but with l-serine biosynthesis were identified from Methylobacterium sp. MB200 mutants. Gene gnd, encoding 6-phosphogluconate dehydrogenase (PGDH), was selected for further evaluation. The gnd deletion mutant showed a 600% increase in d-serine tolerance and an 80% decrease in PGDH activity compared to Methylobacterium sp. MB200. gnd over-expression did not affect d-serine tolerance, whereas it did increase enzyme-activity up to 136%. Additionally, analysis revealed that in Methylobacterium sp. MB200, l-serine inhibited PGDH activity. The deletion of gnd did not affect growth, whereas it did enhance the biosynthesis of l-serine, resulting in a 225% increase in production of l-serine compared to the wild-type.

Conclusion

gnd, one of the five genes identified here that is associated with l-serine synthesis, can be developed as a potential candidate for metabolic engineering to promote l-serine synthesis in Methylobacterium sp. MB200.

Keywords

Methylobacterium l-Serine Metabolic engineering Gene deletion 6-Phosphogluconate dehydrogenase 

Notes

Acknowledgements

This project was supported by the National Nature Science Foundation of China (Grant No. 31160022).

Supporting information

Supplementary Fig. 1—Schematic diagram of construction of the gnd-deleted vector pk18mobSacB-gndLR.

Supplementary Fig. 2—TLC analysis of l-serine synthesis by Methylobacterium sp. MB200 and mutant strains.

Supplementary Fig. 3—PCR verification of the MB200Δgnd, MB200∆gnd(pCM80-gnd), and MB200(pCM80-gnd).

Supplementary Table 1—Bacterial strains, plasmids, and primers used in this study.

Compliance with ethical standards

Conflict of interest

The authors declare no conflict of interest.

Supplementary material

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Supplementary material 1 (TIFF 36 kb)
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Supplementary material 3 (TIFF 8162 kb)
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Supplementary material 4 (DOCX 17 kb)

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Copyright information

© Springer Nature B.V. 2018

Authors and Affiliations

  • Xian Li
    • 1
    • 2
  • Bo Wu
    • 3
  • Kan Zhou
    • 4
  • Chengjian Jiang
    • 1
  • Peihong Shen
    • 1
  1. 1.State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, College of Life Science and TechnologyGuangxi UniversityNanningPeople’s Republic of China
  2. 2.Medical CollegeGuangxi UniversityNanningPeople’s Republic of China
  3. 3.Guangxi University for NationalitiesNanningPeople’s Republic of China
  4. 4.First affiliated hospital of medical CollegeShihezi UniversityShiheziPeople’s Republic of China

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