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Expression of β-glucuronidase on the surface of bacteria enhances activation of glucuronide prodrugs

Cancer Gene Therapy volume 20, pages 276–281 (2013)Cite this article

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Abstract

Extracellular activation of hydrophilic glucuronide prodrugs by β-glucuronidase (βG) was examined to increase the therapeutic efficacy of bacteria-directed enzyme prodrug therapy (BDEPT). βG was expressed on the surface of Escherichia coli by fusion to either the bacterial autotransporter protein Adhesin (membrane βG (mβG)/AIDA) or the lipoprotein (lpp) outermembrane protein A (mβG/lpp). Both mβG/AIDA and mβG/lpp were expressed on the bacterial surface, but only mβG/AIDA displayed enzymatic activity. The rate of substrate hydrolysis by mβG/AIDA-BL21cells was 2.6-fold greater than by pβG-BL21 cells, which express periplasmic βG. Human colon cancer HCT116 cells that were incubated with mβG/AIDA-BL21 bacteria were sensitive to a glucuronide prodrug (p-hydroxy aniline mustard β-D-glucuronide, HAMG) with an half maximal inhibitory concentration (IC50) value of 226.53±45.4 μM, similar to the IC50 value of the active drug (p-hydroxy aniline mustard, pHAM; 70.6±6.75 μM), indicating that mβG/AIDA on BL21 bacteria could rapidly and efficiently convert HAMG to an active anticancer agent. These results suggest that surface display of functional βG on bacteria can enhance the hydrolysis of glucuronide prodrugs and may increase the effectiveness of BDEPT.

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Acknowledgements

This work was supported by grants from the National Research Program for Biopharmaceuticals, National Science Council, Taipei, Taiwan (NSC101-2325-B-037-001, NSC101-2321-B-037-001, NSC101-2313-B-022-001), the Department of Health, Executive Yuan, Taiwan (DOH100-TD-C-111-002) and the Grant of Biosignature in Colorectal Cancers, Academia Sinica, Taiwan.

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  1. C-M Cheng and F M Chen: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Aquaculture, National Kaohsiung Marine University, Kaohsiung, Taiwan

    C-M Cheng

  2. Department of Surgery, Faculty of Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan

    F M Chen & J-Y Wang

  3. Department of Pharmacy, Chia Nan University of Pharmacy and Science, Tainan, Taiwan

    Y-L Lu

  4. Department of Biological Science and Technology, National Chiao Tung University, Hsin-Chu, Taiwan

    S-C Tzou & K-W Liao

  5. Graduate Institute of Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan

    C-H Kao, T-C Cheng & T-L Cheng

  6. Institutes of Basic Medical Sciences, National Cheng Kung University, Tainan, Taiwan

    C-H Chuang

  7. Institute of Biomedical Sciences, Academia Sinica, Taipei, Taiwan

    B-M Chen & S Roffler

  8. Cancer Center, Kaohsiung Medical University Hospital, Kaohsiung, Taiwan

    T-L Cheng

  9. Institute of Biomedical Sciences, National Sun Yat-Sen University, Kaohsiung, Taiwan

    T-L Cheng

  10. Department of Biomedical Science and Environmental Biology, Kaohsiung Medical University, Kaohsiung, Taiwan

    T-L Cheng

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Cheng, CM., Chen, F., Lu, YL. et al. Expression of β-glucuronidase on the surface of bacteria enhances activation of glucuronide prodrugs. Cancer Gene Ther 20, 276–281 (2013). https://doi.org/10.1038/cgt.2013.17

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