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β-Carotene Biosynthesis in Probiotic Bacteria

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Abstract

Susceptibility to deadly diarrheal diseases is partly due to widespread pediatric vitamin A deficiency. To increase vitamin A coverage in malnourished children, we propose to engineer a probiotic bacterium that will produce β-carotene in the intestine, which will be metabolized to vitamin A. Such a therapy has the potential to broadly stimulate mucosal immunity and simultaneously reduce the incidence and duration of diarrheal disease. To that end, a β-carotene-producing variant of the probiotic Escherichia coli strain Nissle 1917 (EcN-BETA) was generated. Notably, the strain produces β-carotene under anaerobic conditions, reflective of the gut environment. EcN-BETA also retains β-carotene production capability after lyophilization, suggesting that it may be amenable to dry formulation. Moreover, EcN-BETA activates murine dendritic cells in vitro, suggesting that the presence of β-carotene may not diminish the immunostimulatory capacity of EcN. Finally, we present a framework through which further improvements may enable approaches such as the one described in this report to yield innovative life-saving therapies for the developing world.

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Acknowledgments

E. coli Nissle 1917 was a generous gift from Dr. Paul Cohen (University of Rhode Island). Several plasmids containing the requisite genes of the β-carotene biosynthesis pathway, including pSTBlue1-BETAipi and pAC-BETA, were generously provided by Dr. Francis Cunningham (University of Maryland). A plasmid containing the human BCDO gene was graciously provided by Prof. Don Zack (Johns Hopkins University). A plasmid containing the mouse BCMO (pBAD/BCD) and a doubly transformed E. coli clone (pBAD/BCD + pAC-BETA) were generously provided by Dr. Michael Redmond (National Eye Institute). Thanks are extended to Dr. Walter Moos (Vice President, SRI Biosciences) and Dr. Amit Galande (Director, SRI Biosciences) for encouragement and support, to Dr. Eugenia Poliankov (National Eye Institute) and Dr. Kevin Ramkissoon (SRI International) for helpful discussions, and to Than-Thuy Tran for technical assistance with the BMDC experiments. This work was supported by a grant from the Bill & Melinda Gates Foundation through the Grand Challenges Explorations Initiative. SRI International’s Center for Advanced Drug Research was established through funding support from the Commonwealth of Virginia.

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The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Biosciences Division, SRI International, Harrisonburg, VA, 22802, USA

    Jennifer K. Miller, Aaron N. Endsley, Krishna Kodukula & Douglas S. Watson

  2. Biosciences Division, SRI International, Menlo Park, CA, 94025, USA

    M. Travis Harrison, Annalisa D’Andrea & Fangfang Yin

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  1. Jennifer K. Miller
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Miller, J.K., Harrison, M.T., D’Andrea, A. et al. β-Carotene Biosynthesis in Probiotic Bacteria. Probiotics & Antimicro. Prot. 5, 69–80 (2013). https://doi.org/10.1007/s12602-013-9133-3

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