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Effects of nitrogen and phosphorus limitation on lipid accumulation by Chlorella kessleri str. UTEX 263 grown in darkness

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Abstract

Growing algae in darkness for biodiesel production eliminates the challenges of evaporation and light penetration reported for open ponds and the costs and fouling that plague photobioreactors. The current study demonstrated that Chlorella kessleri str. UTEX 263 could grow heterotrophically in the dark on pure sugars or lignocellulosic hydrolysates of plant biomass. Hydrolysates of a prairie grass native to Kansas, Big Bluestem (Andropogon gerardii), supported the growth of C. kessleri in the dark. Nitrogen limitation stimulated the accumulation of biodiesel lipids by 10-fold in heterotrophic cultures grown on pure sugars or Big Bluestem hydrolysate. Limiting P in the growth medium also was shown to increase cellular lipid accumulation in C. kessleri. Iron limitation was not sufficient to increase cellular lipid content. Crude biomass extracts may have levels of N that cannot be easily removed, which are high enough to relieve N limitations in growth media. This initial study suggests that P might be more easily removed from biomass extracts than N for increasing cellular lipid production by nutrient limitation and further that native prairie grasses are potentially suitable as sources of lignocellulosic sugars.

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Acknowledgments

The authors appreciate the technical support of James Crisler, Timothy Eberl, Casper Fredsgaard, Devon Miller, and Robert Tolley. We thank Belinda Sturm (University of Kansas) for performing lipid analyses. Biomass hydrolysates were kindly provided by Donghai Wang and Ke Zhang (Kansas State University) and Accelerase enzyme was a gift from Stephen Crawford, Deborah Dodge, and Chris Nguyen (DuPont). Special thanks to Gregory Houseman and Leland Russell for collecting Big Bluestem biomass. Preliminary accounts of this work have been presented previously and abstracted (Dandinpet and Schneegurt 2013a, b, 2014; Shrestha and Schneegurt 2016). This is publication no. 33 in the series of reports from the Wichita State University Biological Field Station.

Funding

This work was supported by an award from Kansas National Science Foundation (NSF) Established Program to Stimulate Competitive Research (EPSCoR) (EPS-0903806). Additional student support was from Kansas Institutional Development Award (IDeA) Networks of Biomedical Research Excellence (KINBRE) of the National Institute of General Medical Sciences (NIGMS) of the National Institutes of Health (NIH) (P20 GM103418).

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  1. Department of Biological Sciences, Wichita State University, Wichita, KS, 67260, USA

    Nayan Shrestha, Kiran K. Dandinpet & Mark A. Schneegurt

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  1. Nayan Shrestha
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Shrestha, N., Dandinpet, K.K. & Schneegurt, M.A. Effects of nitrogen and phosphorus limitation on lipid accumulation by Chlorella kessleri str. UTEX 263 grown in darkness. J Appl Phycol 32, 2795–2805 (2020). https://doi.org/10.1007/s10811-020-02144-x

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