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Assessment of Nannochloropsis gaditana growth and lipid accumulation with increased inorganic carbon delivery

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Abstract

Algal biomass refineries for sustainable transportation fuels, in particular biodiesel, will benefit from algal strain enhancements to improve biomass and lipid productivity. Specifically, the supply of inorganic carbon to microalgal cultures represents an area of great interest due to the potential for improved growth of microalgae and the possibility for incorporation with CO2 mitigation processes. Combinations of bicarbonate (HCO3−) salt addition and application of CO2 to control pH have shown compelling increases in growth rate and lipid productivity of fresh water algae. Here, focus was placed on the marine organism, Nannochloropsis gaditana, to investigate growth and lipid accumulation under various strategies of enhanced inorganic carbon supply. Three gas application strategies were investigated: continuous sparging of atmospheric air, continuous sparging of 5% CO2 during light hours until nitrogen depletion, and continuous sparging of atmospheric air supplemented with 5% CO2 for pH control between 8.0 and 8.3. These gas sparging schemes were combined with addition of low concentrations (5 mM) of sodium bicarbonate at inoculation and high concentration (50 mM) of sodium bicarbonate amendments just prior to nitrogen depletion. The optimum scenario observed for growth of N. gaditana under these inorganic carbon conditions was controlling pH with 5% CO2 on demand, which increased both growth rate and lipid accumulation. Fatty acid methyl esters were primarily comprised of C16:0 (palmitic) and C16:1 (palmitoleic) aliphatic chains. Additionally, the use of high concentration (50 mM) of bicarbonate amendments further improved lipid content (up to 48.6%) under nitrogen deplete conditions when paired with pH-controlled strategies.

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Acknowledgements

A special thank you to all members of the MSU Algal Biofuels Group for their introspective discourse on algal biofuel-related topics.

Funding

A portion of this research was supported by the U.S. Department of Energy (DOE) Office of Energy Efficiency and Renewable Energy (EERE) Biomass Program under Contract No. DE-EE0005993. Support for TCP was also provided by Church and Dwight Co., Inc. Instrumental support was provided through the Environmental and Biofilm Mass Spectrometry Facility at the College of Engineering (COE), and the Center for Biofilm Engineering (CBE), at Montana State University (MSU).

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Authors and Affiliations

  1. Department of Chemical and Biological Engineering, and Center for Biofilm Engineering, Montana State University, Bozeman, MT, 59717, USA

    Todd C. Pedersen, Robin Gerlach & Brent M. Peyton

  2. Department of Bioproducts and Biosystems Engineering, University of Minnesota, St. Paul, MN, 55108, USA

    Robert D. Gardner

Authors
  1. Todd C. Pedersen
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  2. Robert D. Gardner
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  3. Robin Gerlach
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  4. Brent M. Peyton
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Correspondence to Brent M. Peyton.

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Competing interests

A patent entitled “Bicarbonate Trigger for Inducing Lipid Accumulation in Algal Systems” (Pat. No 9,096,875) was co-authored by contributing authors Robert D. Gardner and Brent M. Peyton.

Electronic supplementary material

Electronic Supplementary Fig. 1

Growth [cells mL-1] (a), pH (b), nitrate concentration [mg NO3--N L-1] (c), and total chlorophyll [mg L-1] (d) of cultures of N. gaditana grown using atmospheric air sparge. Air-1: Continuous sparge of atmospheric air (△). Air-2: Continuous sparge of atmospheric air and 50mM NaHCO3 just prior to nitrogen depletion (○). Air-3: Continuous sparge of atmospheric air with 5mM initial NaHCO3 and an additional 50mM NaHCO3 just prior to nitrogen depletion (□). Time of nitrate depletion is indicated in Table 2. Error bars represent ±95% CI (n=3) (JPEG 310 kb)

High resolution image (EPS 2017 kb)

Electronic Supplementary Fig. 2

Growth [cells mL-1] (a), pH (b), nitrate concentration [mg NO3--N L-1] (c), and total chlorophyll [mg L-1] (d) of cultures of N. gaditana cultured with CO2. CO2-1: Continuous sparge of atmospheric air supplemented with 5% CO2 (v/v) during daylight hours until nitrogen depletion (◇). CO2-2: Continuous sparge of atmospheric air supplemented periodically with 5% CO2 (v/v) to control pH (8.0 to 8.3) (○). Time of nitrate depletion is indicated in Table 2. Error bars represent ±95% CI (n=3) (JPEG 290 kb)

High resolution image (EPS 1933 kb)

Electronic Supplementary Fig. 3

Growth [cells mL-1] (a), pH (b), nitrate concentration [mg NO3--N L-1] (c), and total chlorophyll [mg L-1] (d) of cultures of N. gaditana cultured with CO2 and bicarbonate. CO2/HCO3-1: pH control (8.0 to 8.3) and supplemented with 50mM NaHCO3 just prior to nitrogen depletion (△). CO2/HCO3-2 and -3: pH control (8.0 to 8.3) with 5mM initial NaHCO3 and an additional 50mM NaHCO3 just prior to nitrogen depletion (○ and ◇). Time of nitrate depletion is indicated in Table 2. Error bars represent ±95% CI (n=3) (JPEG 287 kb)

High resolution image (EPS 1936 kb)

Electronic Supplementary Fig. 4

Growth [cells mL-1] (a), pH (b), nitrate concentration [mg NO3--N L-1] (c), and total chlorophyll [mg L-1] (d) of cultures of N. gaditana cultured with CO2 and bicarbonate. CO2/HCO3-4: 5% CO2 (v/v) during daylight hours until nitrogen depletion and supplemented with 5mM initial NaHCO3 (□). CO2/HCO3-5: 5% CO2 (v/v) during daylight hours until nitrogen depletion and supplemented with 50mM NaHCO3 just prior to nitrogen depletion (×). Time of nitrate depletion is indicated in Table 2. Error bars represent ±95% CI (n=3) (JPEG 251 kb)

High resolution image (EPS 1852 kb)

Supplementary Table 1

Estimated alkalinity considering nitrate consumption during each study, initial bicarbonate supplementation, and bicarbonate supplementation at nitrate depletion (DOCX 20.7 kb)

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Pedersen, T.C., Gardner, R.D., Gerlach, R. et al. Assessment of Nannochloropsis gaditana growth and lipid accumulation with increased inorganic carbon delivery. J Appl Phycol 30, 2155–2166 (2018). https://doi.org/10.1007/s10811-018-1470-x

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  • DOI: https://doi.org/10.1007/s10811-018-1470-x

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