Synthetic cell–cell communication in a three-species consortium for one-step vitamin C fermentation

  • En-Xu Wang
  • Yu Liu
  • Qian Ma
  • Xiu-Tao Dong
  • Ming-Zhu DingEmail author
  • Ying-Jin Yuan
Original Research Paper



A three-species consortium for one-step fermentation of 2-keto-l-gulonic acid (2-KGA) was constructed to better strengthen the cell–cell communication. And the programmed cell death module based on the LuxI/LuxR quorum-sensing (QS) system was established in Gluconobacter oxydans to reduce the competition that between G. oxydans and Ketogulonicigenium vulgare.


By constructing and optimizing the core region of the promoter, which directly regulated the expression of lethal ccdB genes in QS system, IR3C achieved the best lethal effect. The consortium of IR3C- K. vulgareBacillus megaterium (abbreviated as 3C) achieved the highest 2-KGA titer (68.80 ± 4.18 g/l), and the molar conversion rate was 80.7% within 36 h in 5 l fermenter. Metabolomic analysis on intracellular small molecules of consortia 3C and 1C showed that most amino acids (such as glycine, leucine, methionine and proline) and TCA cycle intermediates (such as succinic acid, fumaric acid and malic acid) were significantly affected. These results further validated that the programmed cell death module based on the LuxI/LuxR QS system in G. oxydans could also faciliate better growth and higher production of consortium 3C for one-step fermentation.


We successfully constructed a novel three-species consortia for one-step vitamin C fermentation by strengthening the cell–cell communication. This will be very useful for probing the rational design principles of more complex multi-microbial consortia.


One-step fermentation Programmed cell death Quorum-sensing 2-keto-l-gulonic acid 



This work was funded by the National Key Research and Development Program of China (Grant Nos. 2018YFA0902103, 2018YFA0902200), National Natural Science Foundation of China (Grant Nos. 21676190, 21621004), Innovative Talents and Platform Program of Tianjin (Grant No. 16PTGCCX00140).

Supporting information

Supplementary Table 1— Primers for the synthesis of IR1C

Supplementary Table 2— Primers for qPCR reaction

Supplementary Fig. 1— Flow chart of the construction of the gene circuit

Supplementary Fig. 2—Construction of a synthetic microbial consortium of G. oxydans- K. vulgare- Bacillus megaterium for cell-cell communication in one-step vitamin C precursor fermentation

Supplementary Fig. 3—Relationship between OD600 and dry cell weight (DCW) of G. oxydans

Additional file 1—IR1C is split into full sequence of 4 Building Blocks

Supplementary material

10529_2019_2705_MOESM1_ESM.docx (340 kb)
Supplementary material 1 (DOCX 339 kb)


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Copyright information

© Springer Nature B.V. 2019

Authors and Affiliations

  • En-Xu Wang
    • 1
    • 2
  • Yu Liu
    • 1
    • 2
  • Qian Ma
    • 1
    • 2
  • Xiu-Tao Dong
    • 1
    • 2
  • Ming-Zhu Ding
    • 1
    • 2
    Email author
  • Ying-Jin Yuan
    • 1
    • 2
  1. 1.Frontier Science Center for Synthetic Biology and Key Laboratory of Systems Bioengineering (Ministry of Education), School of Chemical Engineering and TechnologyTianjin UniversityTianjinPeople’s Republic of China
  2. 2.Collaborative Innovation Center of Chemical Science and Engineering (Tianjin)Tianjin UniversityTianjinPeople’s Republic of China

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