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Monitoring and dynamically controlling glucose uptake rate and central metabolism

Nature Chemical Engineering volume 2, pages 50–62 (2025)Cite this article

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Abstract

The rate of glucose import directly affects the maximum possible flux of central carbon metabolism. However, few tools can directly monitor the cellular glucose uptake rate. Here we report the development of a set of programmable bifunctional glucose uptake rate biosensors (GURBs) for real-time monitoring of glucose uptake rate, which enable the dynamic activation and inhibition of glucose uptake and central metabolism in Escherichia coli. These genetic circuits are used to monitor the glucose uptake rates of strains under different culture conditions. Also, feedback-loop control systems are designed to make cells rely on the glucose uptake rate to tune the target metabolic modules, resulting in a substantial increase of the titers of l-tryptophan, riboflavin and d-lactic acid. The glucose-uptake-rate-responsive genetic circuits developed in this study will serve as an effective tool for the dynamic control of glucose uptake and central metabolism.

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Fig. 1: Design of a glucose uptake rate biosensor.
Fig. 2: Construction and characterization of GURB variants that respond positively to the glucose uptake rate.
Fig. 3: Quantitative analysis of GURB28 responses.
Fig. 4: Glucose uptake rates of strains measured under different culture conditions using the online biosensor and offline residual glucose analyzer.
Fig. 5: Enhanced production of l-Trp biosynthesis in E. coli by dynamic regulation using the GURB system.
Fig. 6: Enhanced production of riboflavin, and d-lactic acid biosynthesis in E. coli by dynamic regulation using the GURB system.

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Data availability

The main data supporting the findings of this study are available within the Article and its Supplementary Information. Specific data P values are included in the source data files. Additional details on the datasets and protocols that support the findings of this study will be made available by the corresponding author upon request. Data are provided at https://doi.org/10.6084/m9.figshare.24790872 (ref. 71). Source data are provided with this paper.

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Acknowledgements

S.Y.L. served as the chairman of the scientific advisory board of the Tianjin Institute of Industrial Biotechnology. D.D. was supported by the National Key R&D Program of China (2021YFC2100900), National Nature Science Foundation of China (32100062), the Postdoctoral Science Foundation of China (2021M703438) and Tianjin Synthetic Biotechnology Innovation Capacity Improvement Project (TSBICIP-CXRC-029). D.Z. was supported by the National Science Fund for Distinguished Young Scholars (22325807), the National Nature Science Foundation of China (22178372) and the Tianjin Synthetic Biotechnology Innovation Capacity Improvement Project (TSBICIP-CXRC-055). S.Y.L. was supported by the Development of Platform Technologies of Microbial Cell Factories for the Next-Generation Biorefineries Project (2022M3J5A1056117) of the National Research Foundation supported by the Korean Ministry of Science and ICT. We thank T. Chen for plasmid p20C-Bsrib and X. Zhang for strain Lac04. We thank J. Shen for SPR technology assistance, Z. Zhang for MS contributions and L. Qi for flow cytometry expertise.

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  1. These authors contributed equally: Dongqin Ding, Yaru Zhu.

Authors and Affiliations

  1. Tianjin Institutes of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin, P. R. China

    Dongqin Ding, Yaru Zhu, Danyang Bai, Tongxin Wan & Dawei Zhang

  2. National Technology Innovation Center of Synthetic Biology, Tianjin, P. R. China

    Dongqin Ding, Yaru Zhu, Danyang Bai & Dawei Zhang

  3. University of Chinese Academy of Sciences, Beijing, P. R. China

    Dongqin Ding & Dawei Zhang

  4. Department of Chemical and Biomolecular Engineering (BK21 four program), Korea Advanced Institute of Science and Technology (KAIST), 291 Daehak-ro, Yuseong-gu, Daejeon, Republic of Korea

    Sang Yup Lee

  5. College of Biotechnology Engineering, Tianjin University of Science and Technology, Tianjin, P. R. China

    Dawei Zhang

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Contributions

D.D., SY.L. and D.Z. initiated the project and designed the experiments. D.D., Y.Z., D.B. and T.W. performed the experiments. D.D., Y.Z., SY.L. and D.Z. wrote the manuscript.

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Correspondence to Sang Yup Lee or Dawei Zhang.

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Ding, D., Zhu, Y., Bai, D. et al. Monitoring and dynamically controlling glucose uptake rate and central metabolism. Nat Chem Eng 2, 50–62 (2025). https://doi.org/10.1038/s44286-024-00163-w

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