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Novel Production of Terpenoids in Escherichia coli and Activities Against Breast Cancer Cell Lines

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Abstract

Metabolic engineering of heterologous pathways has allowed the production of therapeutically important compounds in microbial systems. Here, we report the engineering of a monoterpenoid biosynthetic pathway into Escherichia coli. Five genes encoding sequential enzymes for perillyl alcohol biosynthesis from the precursors isopentenyl pyrophosphate (IPP) and dimethylallyl pyrophosphate (DMAPP) were engineered into E. coli. Expression of these genes allowed the production of the intermediate limonene, but the downstream monoterpenoid, perillyl alcohol, was not detected. A new compound was detected but could not be identified based on the data obtained. Only 1.6 μg/ml of the compound was being produced from the engineered E. coli strain, but, when these cultures were fed limonene as a substrate, the production was nearly 250 μg/ml. This unknown compound inhibited the cell proliferation of MCF-7 and MDA-MB-231 breast cancer cells in 48-h treatment experiments. This compound may have potential benefits in breast cancer treatment. This is the first report showing the production of a monoterpenoid in engineered E. coli and its antiproliferative effects in breast cancer cells.

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Acknowledgments

We acknowledge and thank the Department of Biological Sciences and the College of Graduate Studies, University of the Sciences, for the support of this project. We thank Dr. Catherine Moore and Dr. Natalia Coleman for generously providing the MDA-MB-231 cells and MCF-7 cells, respectively. We also thank Mr. Carl Walasek for his assistance with the statistical analyses.

Conflict of Interest

The authors declare no competing interests or conflicts of interest.

Author’s Contribution

SG, MGM, JRA, and JRP designed experiments, interpreted results, and participated in preparation of the manuscript; the experiments were conducted primarily by SG. FS participated in the design and interpretation of the GC-MS experiments. VAM and FS participated in revision of the manuscript. All authors agree with the findings and interpretations contained in the manuscript and agree to the submission to Applied Biochemistry and Biotechnology.

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Author notes

  1. Shweta Gupta

    Present address: Molecular and Behavioral Neuroscience Institute, University of Michigan, 205 Zina Pitcher Pl, Ann Arbor, MI, 48108, USA

  2. Vandana A. Miller

    Present address: A. J. Drexel Plasma Institute, 3120-24 Market St, Philadelphia, PA, 19104, USA

  3. Jennifer R. Anthony

    Present address: Department of Science, Technology, Engineering & Mathematics, Delaware County Community College, 915 S. Media Line Road, Pennsylvania, PA, 19063, USA

Authors and Affiliations

  1. Department of Biological Sciences, University of the Sciences, 600 South Forty-Third Street, Philadelphia, PA, 19104, USA

    Shweta Gupta, Melissa G. Marko, Frederick T. Schaefer, Jennifer R. Anthony & John R. Porter

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  1. Shweta Gupta
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  2. Melissa G. Marko
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Correspondence to John R. Porter.

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Gupta, S., Marko, M.G., Miller, V.A. et al. Novel Production of Terpenoids in Escherichia coli and Activities Against Breast Cancer Cell Lines. Appl Biochem Biotechnol 175, 2319–2331 (2015). https://doi.org/10.1007/s12010-014-1382-4

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  • DOI: https://doi.org/10.1007/s12010-014-1382-4

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