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Applying Statistical Design of Experiments (DOE) To Understanding the Effect of Growth Media Components on Cupriavidus necator H16 Growth

Lookup NU author(s): Christopher Azubuike, Dr Martin EdwardsORCiD, Professor Angharad MR GatehouseORCiD, Dr Thomas HowardORCiD

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This work is licensed under a Creative Commons Attribution 4.0 International License (CC BY 4.0).

Abstract

Cupriavidus necator H16 is gaining significant attention as a microbial chassis for range of biotechnological applications. Whilst the bacterium is a major producer of bioplastics, its lithoautotrophic and versatile metabolic capabilities make the bacterium a promising microbial chassis for biofuels and chemicals using renewable resources. It remains necessary to develop appropriate experimental resources to permit controlled bioengineering and system optimization of this microbe. Here we employed statistical design of experiments to gain understanding of the impact of components of defined media on C. necator growth and built a model that can predict the bacterium's cell density based on media components. This highlighted media components, and interaction between components, having the most effect on growth: fructose, amino acids, trace elements, CaCl₂ and Na₂HPO₄ contributed significantly to growth (t < -1.65 or > 1.65); copper and histidine were found to interact and must be balanced for robust growth. Our model was experimentally validated and found to correlate well, r² = 0.85. Model validation at large culture scales showed correlations between our model predicted growth ranks and experimentally determined ranks at 100 mL in shake flasks (ρ = 0.87) and 1 L in bioreactor (ρ = 0.90). Our approach provides valuable and quantifiable insights on the impact of media components on cell growth and can be applied to model other C. necator responses that are crucial for its deployment as a microbial chassis. This approach can be extended to other non-model microbes of medical and industrial biotechnological importance.

Publication metadata

Author(s): Azubuike CC, Edwards MG, Gatehouse AMR, Howard TP

Publication type: Article

Publication status: Published

Journal: Applied and Environmental Microbiology

Year: 2020

Volume: 86

Issue: 17

Online publication date: 19/06/2020

Acceptance date: 31/05/2020

Date deposited: 16/07/2020

ISSN (print): 0099-2240

ISSN (electronic): 1098-5336

Publisher: American Society for Microbiology

URL: https://doi.org/10.1128/AEM.00705-20

DOI: 10.1128/AEM.00705-20

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