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Enhanced production of hairy root metabolites using microbubble generator

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Abstract

Previously, increased partitioning of the natural product nicotine from tobacco hairy roots into the culture media was achieved by altering the expression of the nicotine uptake permease gene. The present study demonstrated that further increases in nicotine yield in the media were attained by using surfactant-stabilized microbubbles. Compared to other non-ionic surfactants (Tween 20 and Tween 80) and the ionic surfactant SDS, Triton X-100 (TX100) both increased total nicotine production and exudation into the hairy root culture media. In comparison to surfactant-free medium, TX100 at 10, 25, and 50 mg l−1 did not show strong inhibition of hairy root growth. At 4,000 rpm shear speed, microbubbles stabilized by 10, 25, and 50 mg l−1 TX100 had k L a of 22.3, 36.2, and 44.1 h−1 in Gamborg’s B5 medium, respectively, in comparison to 16.4 h−1 with conventional air sparging. In a 1-l bioreactor, microbubbles stabilized by TX100 were applied to hairy roots after the inoculated root tips were self-immobilized by branching. With microbubble dispersion, dissolved oxygen rapidly increased from 60 to 85 %, and hairy root growth rate increased. Nicotine accumulation in culture medium with microbubbles reached 146 mg l−1 after 30 days cultivation. These results show that combining genetic modification with surfactant-stabilized microbubble dispersion can substantially increase levels of nicotine in the media of hairy root cultures.

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Abbreviations

MBD:

Microbubble dispersion

NUP:

Nicotine uptake permease

TX100:

Triton X-100

vvm:

Volume of gas per volume of liquid per minute

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Acknowledgments

This work was supported by the United States Department of Agriculture NIFA award 2009-34602-20015 to the Virginia Tech Biodesign and Bioprocessing Research Center, and the USTAR Synthetic Bioproducts Center.

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Authors and Affiliations

  1. Department of Biological Engineering, USTAR Bioenergy Center, Utah State University, Logan, UT, 84341, USA

    Bo Zhao & Foster A. Agblevor

  2. Department of Plant Pathology, Physiology, and Weed Science, Virginia Tech, Blacksburg, VA, 24061, USA

    John G. Jelesko

Authors
  1. Bo Zhao
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  2. Foster A. Agblevor
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  3. John G. Jelesko
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Corresponding authors

Correspondence to Bo Zhao or Foster A. Agblevor.

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Bo Zhao and Foster A. Agblevor are co-communicating authors.

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Zhao, B., Agblevor, F.A. & Jelesko, J.G. Enhanced production of hairy root metabolites using microbubble generator. Plant Cell Tiss Organ Cult 117, 157–165 (2014). https://doi.org/10.1007/s11240-014-0428-1

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  • DOI: https://doi.org/10.1007/s11240-014-0428-1

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