Abstract
Objectives
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.
Results
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. vulgare–Bacillus 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.
Conclusions
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.
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Acknowledgements
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
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Wang, EX., Liu, Y., Ma, Q. et al. Synthetic cell–cell communication in a three-species consortium for one-step vitamin C fermentation. Biotechnol Lett 41, 951–961 (2019). https://doi.org/10.1007/s10529-019-02705-2
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DOI: https://doi.org/10.1007/s10529-019-02705-2