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

C. M. Cheng, F. M. Chen, Y. L. Lu, Shey-Cherng Tzou, J. Y. Wang, C. H. Kao, Kuang-Wen Liao, T. C. Cheng, C. H. Chuang, B. M. Chen, S. Roffler*, T. L. Cheng

*Corresponding author for this work

Research output: Contribution to journalArticle

10 Scopus citations

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 IC 50 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.

Original languageEnglish
Pages (from-to)276-281
Number of pages6
JournalCancer Gene Therapy
Volume20
Issue number5
DOIs
StatePublished - 1 May 2013

Keywords

  • Autotransporter protein adhesin
  • Bacteria-directed enzyme prodrug therapy
  • Extracellular activation
  • Surface display
  • β-glucuronidase

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    Cheng, C. M., Chen, F. M., Lu, Y. L., Tzou, S-C., Wang, J. Y., Kao, C. H., Liao, K-W., Cheng, T. C., Chuang, C. H., Chen, B. M., Roffler, S., & Cheng, T. L. (2013). Expression of β-glucuronidase on the surface of bacteria enhances activation of glucuronide prodrugs. Cancer Gene Therapy, 20(5), 276-281. https://doi.org/10.1038/cgt.2013.17