Biotechnology and Bioprocess Engineering

, Volume 24, Issue 1, pp 264–271 | Cite as

Prescreening of Natural Products in Drug Discovery Using Recombinant Bioluminescent Bacteria

  • Eui Jong Kim
  • Ho Bin SeoEmail author
  • Man Bock GuEmail author
Research Paper


Strains of recombinant bioluminescent bacteria (RBB) which respond to toxic environments using various stress promoters are practical means of assessing toxicity. In previous research, RBB has proven useful for highthroughput screening in the drug development process. The goal of this research is to demonstrate that RBB can also be used for the toxicity screening of natural products. The RBB strains used were DPD2511, BBTSbmC, TV1061, and GC2, which were selected to respond to oxidative stress, DNA damage, protein damage, and cellular toxicity, respectively. The test drugs (paclitaxel, etoposide, and pentostatin) were carefully selected because these drugs needed to be natural products or their derivatives whose cellular toxicity had previously been reported from human cell line assays. After treating the RBB strains with various doses of the chosen drugs, their bioluminescent signals were measured over time. The effectiveness of the RBB method was proven by comparing its results to existing toxicity data for the selected drugs. In addition, a similar test using podophyllotoxin, a precursor of etoposide, and a derivative of podophyllotoxin, teniposide, was conducted to prove that the RBB method is suitable for a comparative analysis of toxicity among chemicals with similar molecular structures. As a detection method, RBB bacteria provide a much easier and more rapid culturing process compared to conventional human cell line assays. Because the implementation of the RBB method in the drug discovery process would enable efficient prescreening, a significant reduction in time, effort, and development costs are expected.


recombinant bioluminescent bacteria (RBB) drug discovery natural products prescreening 


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  1. 1.
    Butler, M. S. (2008) Natural products to drugs: natural productderived compounds in clinical trials. Natural Product Reports 25: 475–516.CrossRefGoogle Scholar
  2. 2.
    Newman, D. J. and G. M. Cragg (2007) Natural products as sources of new drugs over the last 25 years. Journal of Natural Products 70: 461–477.CrossRefGoogle Scholar
  3. 3.
    Harvey, A. L. (2008) Natural products in drug discovery. Drug Discovery Today 13: 894–901.CrossRefGoogle Scholar
  4. 4.
    Li, J. W. H. and J. C. Vederas (2009) Drug discovery and natural products: end of an era or an endless frontier? Science 325: 161–165.CrossRefGoogle Scholar
  5. 5.
    Rishton, G. M. (2008) Natural products as a robust source of new drugs and drug leads: past successes and present day issues. American Journal of Cardiology 101: S43–S49.Google Scholar
  6. 6.
    Lam, K. S. (2007) New aspects of natural products in drug discovery. Trends in Microbiology 15: 279–289.CrossRefGoogle Scholar
  7. 7.
    Bunel, V., M. Ouedraogo, A. T. Nguyen, C. Stévigny, and P. Duez (2014) Methods applied to the in vitro primary toxicology testing of natural products: state of the art, strengths, and limits. Planta Medica 80: 1210–1226.CrossRefGoogle Scholar
  8. 8.
    Van Dyk, T. K., D. R. Smulski, T. R. Reed, S. H. I. M. S. H. O. N. Belkin, A. C. Vollmer, and R. A. LaRossa (1995) Responses to toxicants of an Escherichia coli strain carrying a uspA′:: lux genetic fusion and an E. coli strain carrying a grpE′:: lux fusion are similar. Applied and Environmental Microbiology. 61: 4124–4127.Google Scholar
  9. 9.
    Van Dyk, T. K., T. R. Reed, A. C. Vollmer, and R. A. LaRossa (1995) Synergistic induction of the heat shock response in Escherichia coli by simultaneous treatment with chemical inducers. Journal of Bacteriology 177: 6001–6004.CrossRefGoogle Scholar
  10. 10.
    Ahn, J. M., J. H. Kim, J. H. Kim, and M. B. Gu (2010) Randomly distributed arrays of optically coded functional microbeads for toxicity screening and monitoring. Lab on a Chip. 10: 2695–2701.CrossRefGoogle Scholar
  11. 11.
    Jung, I., H. B. Seo, J. E. Lee, B. C. Kim, and M. B. Gu (2014) A dip-stick type biosensor using bioluminescent bacteria encapsulated in color-coded alginate microbeads for detection of water toxicity. Analyst 139: 4696–4701.CrossRefGoogle Scholar
  12. 12.
    Elad, T., H. B. Seo, S. Belkin, and M. B. Gu (2015) Highthroughput prescreening of pharmaceuticals using a genomewide bacterial bioreporter array. Biosensors and Bioelectronics 68: 699–704.CrossRefGoogle Scholar
  13. 13.
    Rogowsky, P. M., T. J. Close, J. A. Chimera, J. J. Shaw, and C. I. Kado (1987) Regulation of the vir genes of Agrobacterium tumefaciens plasmid pTiC58. Journal of Bacteriology 169: 5101–5112.CrossRefGoogle Scholar
  14. 14.
    LaRossa, R. A. (1998) Bioluminescence: Methods and Protocols., pp. 85–95. Totowa: Humana Press.CrossRefGoogle Scholar
  15. 15.
    Choi, S. H. and M. B. Gu (2001) Phenolic toxicity—detection and classification through the use of a recombinant bioluminescent Escherichia coli. Environmental Toxicology and Chemistry. 20: 248–255.Google Scholar
  16. 16.
    Ahn, J. M., E. T. Hwang, C. H. Youn, D. L. Banu, B. C. Kim, J. H. Niazi, and M. B. Gu (2009) Prediction and classification of the modes of genotoxic actions using bacterial biosensors specific for DNA damages. Biosensors and Bioelectronics 25: 767–772.CrossRefGoogle Scholar
  17. 17.
    Belkin, S., D. R. Smulski, A. C. Vollmer, T. K. Van Dyk, and R. A. LaRossa (1996) Oxidative stress detection with Escherichia coli harboring a katG′:: lux fusion. Applied and Environmental Microbiology 62: 2252–2256.Google Scholar
  18. 18.
    Gu, M. B. and G. C. Gil (2001) A multi-channel continuous toxicity monitoring system using recombinant bioluminescent bacteria for classification of toxicity. Biosensors and Bioelectronics 16: 661–666.CrossRefGoogle Scholar
  19. 19.
    Van Dyk, T. K., W. R. Majarian, K. B. Konstantinov, R. M. Young, P. S. Dhurjati, and R. A. Larossa (1994) Rapid and sensitive pollutant detection by induction of heat shock genebioluminescence gene fusions. Applied and Environmental Microbiology. 60: 1414–1420.Google Scholar
  20. 20.
    Marincs, F. and D. W. White (1994) Immobilization of Escherichia coli expressing the lux genes of Xenorhabdus luminescens. Applied and Environmental Microbiology 60: 3862–3863.Google Scholar
  21. 21.
    Budavari, S., M. J. O’Neil, and A. Smith (1989) The Merck Index: an encyclopedia of chemicals, drugs, and biological. Rahway, New Jersey, Merck and Co. Inc PMid, 2666071.Google Scholar
  22. 22.
    Grem, J. L., K. D. Tutsch, K. J. Simon, D. B. Alberti, J. K. Willson, D. C. Tormey, S. Swaminathan, and D. L. Trump (1987) Phase I study of taxol administered as a short iv infusion daily for 5 days. Cancer Treatment Reports 71: 1179–1184.Google Scholar
  23. 23.
    Meshkini, A. and R. Yazdanparast (2012) Involvement of oxidative stress in taxol-induced apoptosis in chronic myelogenous leukemia K562 cells. Experimental and Toxicologic Pathology 64: 357–365.CrossRefGoogle Scholar
  24. 24.
    Fleming, R. A., A. A. Miller, and C. F. Stewart (1989) Etoposide: an update. Clinical Pharmacy 8: 274–293.Google Scholar
  25. 25.
    Hande, K. R. (1998) Etoposide: four decades of development of a topoisomerase II inhibitor. European Journal of Cancer 34: 1514–1521.CrossRefGoogle Scholar
  26. 26.
    Dillman, R. O., R. Mick, and O. R. McIntyre (1989) Pentostatin in chronic lymphocytic leukemia: a phase II trial of Cancer and Leukemia group B. Journal of Clinical Oncology 7: 433–438.CrossRefGoogle Scholar
  27. 27.
  28. 28.
    Oh, S. Y., Y. W. Sohn, J. W. Park, H. J. Park, H. M. Jeon, T. K. Kim, J. S. Lee, J. E. Jung, X. Jin, Y. G. Chung, Y. K. Choi, S. You, J. B. Lee, and H. Kim (2007) Selective cell death of oncogenic Akt-transduced brain cancer cells by etoposide through reactive oxygen species-mediated damage. Molecular Cancer Therapeutics 6: 2178–2187.CrossRefGoogle Scholar
  29. 29.
    Branham, M. T., S. B. Nadin, L. M. Vargas-Roig, and D. R. Ciocca (2004) DNA damage induced by paclitaxel and DNA repair capability of peripheral blood lymphocytes as evaluated by the alkaline comet assay. Mutation Research/Genetic Toxicology and Environmental Mutagenesis 560: 11–17.CrossRefGoogle Scholar
  30. 30.
    Johnston, J. B., A. Begleiter, L. Pugh, M. K. Leith, J. A. Wilkins, D. J. Cavers, and L. G. Israels (1986) Biochemical changes induced in hairy-cell leukemia following treatment with the adenosine deaminase inhibitor 2′-deoxycoformycin. Cancer Research. 46(4 Part 2): 2179–2184.Google Scholar
  31. 31.
    Walles, S. A., R. Zhou, and E. Liliemark (1996) DNA damage induced by etoposide; a comparison of two different methods for determination of strand breaks in DNA. Cancer Letters 105: 153–159.CrossRefGoogle Scholar

Copyright information

© The Korean Society for Biotechnology and Bioengineering and Springer 2019

Authors and Affiliations

  1. 1.Division of LifesciencesKorea UniversitySeoulKorea
  2. 2.Institute of Advanced TechnologySuwonKorea
  3. 3.Department of BiotechnologyKorea UniversitySeoulKorea

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