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Production of biomass and filamentous hemagglutinin by Bordetella bronchiseptica

Bioprocess and Biosystems Engineering volume 37, pages 115–123 (2014)Cite this article

Abstract

The mammalian pathogen Bordetella bronchiseptica was grown under controlled batch conditions with glutamate as the primary carbon and nitrogen source. First, a Box-Behnken statistical design quantified the effect of Mg, sulfate, and nicotinate on the antigen filamentous hemagglutinin (FHA) formation. Using lactic acid as a secondary carbon source for pH control, Mg, and SO4 each negatively affected antigen expression, while nicotinate positively affected antigen expression. Sulfate had a stronger negative effect than Mg with 10 mM eliminating FHA altogether; the highest FHA expression (about 1,000 ng/mL) occurred when either Mg concentration or SO4 concentration, but not both, was about 0.1 mM. Using two Mg and SO4 compositions modeled to yield the greatest antigen expression, three other organic acids were compared as the secondary carbon source: acetate, citrate, and succinate. Mixtures of acetate and glutamate resulted in the greatest organic acid consumption, OD, and FHA concentration (about 1,500 ng/mL), although significant acetate accumulated during these batch processes. The mechanism leading to elevated FHA expression when acetate is the secondary carbon source is unknown, particularly since these cultures were most prone to phase shift to Bvg− cultures.

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Abbreviations

AG:

Cultivations which contained acetate and glutamate as the carbon sources

Bvg:

Bordetella virulence gene regulon

CDM:

Chemically defined medium

CG:

Cultivations which contained citrate and glutamate as the carbon sources

DO:

Dissolved oxygen concentration as a percent of saturation

FHA:

Adhesin filamentous hemagglutinin

L, M, H:

Low, medium, and high concentrations of medium components studied

LG:

Cultivations which contained lactate and glutamate as the carbon sources

OD:

Optical density, a measurement of cell density and growth

SG:

Cultivations which contained succinate and glutamate as the carbon sources

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Acknowledgments

The authors thank Sarah Lee for analytical assistance.

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Authors and Affiliations

  1. Merial Limited, Athens, GA, 30605, USA

    Scott D. Guetter

  2. Biochemical Engineering, College of Engineering, University of Georgia, Athens, GA, 30602, USA

    Mark A. Eiteman

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Correspondence to Mark A. Eiteman.

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Guetter, S.D., Eiteman, M.A. Production of biomass and filamentous hemagglutinin by Bordetella bronchiseptica . Bioprocess Biosyst Eng 37, 115–123 (2014). https://doi.org/10.1007/s00449-013-0977-4

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