Applied Microbiology and Biotechnology

, Volume 98, Issue 11, pp 5261–5273 | Cite as

Mineral and non-carbon nutrient utilization and recovery during sequential phototrophic-heterotrophic growth of lipid-rich algae

  • Pavlo BohutskyiEmail author
  • Kexin Liu
  • Ben A. Kessler
  • Thomas Kula
  • Yongseok Hong
  • Edward J. Bouwer
  • Michael J. Betenbaugh
  • F. C. Thomas Allnutt
Bioenergy and biofuels


A critical factor in implementing microalgal biofuels for mass production is the nutrient requirements. The current study investigated the fate of macro- and micronutrients and their availability in a sequential phototrophic-heterotrophic production process for the lipid rich microalga Auxenochlorella protothecoides. More than 99 % (by weight) of overall process nutrients were supplied during the initial photoautotrophic stage reflecting its significantly larger volume. Under photoautotrophic growth conditions only 9–35 % of supplied Mn, S, Fe, N, Mg, and Cu and less than 5 % of P, Mo, Co, B, Zn, and Ca were consumed by the algae. The rest of these nutrients remain in the spent growth media during the culture concentration-down from an 800 L phototrophic pond to a 5 L heterotrophic fermenter. In contrast, Zn, Mo, Mn, Mg, Ca, and N were exhausted (90–99 % removal) during the first 25 h of the heterotrophic growth stage. The depletion of these key nutrients may have ultimately limited the final biomass density and/or lipid productivity achieved. Approximately 10–20 % of the total supplied S, Mn, Fe, N, and Cu and 5 % of Ca and Zn were assimilated into algal biomass. Several elements including N, P, Mn, B, Cu, Ca, Mg, S, and Fe were released back into the liquid phase by anaerobic digestion (AD) of the residual biomass after lipid extraction. The nutrients recovered from the AD effluent and remaining in the spent medium should be recycled or their initial concentration to the phototrophic stage decreased to enhance process economics and sustainability for future commercialization of algal-derived biofuels.


Sequential mixed trophic state process Fertilizer consumption and recycling Spent growth medium Lipid-extracted algae Anaerobic digestion effluent 



The authors gratefully acknowledge financial support from U.S. DOE CCS Program (Grant No. DE-FE0001888 to Phycal), U.S. NSF CBET Program (Grant No.1236691 to JHU), and the Bureau of Education and Cultural Affairs of U.S. Department of State through an International Fulbright Science and Technology Award to Pavlo Bohutskyi.

Supplementary material

253_2014_5655_MOESM1_ESM.pdf (228 kb)
ESM 1 (PDF 228 kb)


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Copyright information

© Springer-Verlag Berlin Heidelberg 2014

Authors and Affiliations

  • Pavlo Bohutskyi
    • 1
    Email author
  • Kexin Liu
    • 1
  • Ben A. Kessler
    • 2
  • Thomas Kula
    • 2
    • 3
  • Yongseok Hong
    • 1
    • 4
  • Edward J. Bouwer
    • 1
  • Michael J. Betenbaugh
    • 5
  • F. C. Thomas Allnutt
    • 2
    • 6
  1. 1.Department of Geography and Environmental EngineeringJohns Hopkins UniversityBaltimoreUSA
  2. 2.Phycal Inc.Highland HeightsUSA
  3. 3.NPA CoatingsClevelandUSA
  4. 4.Daegu UniversityGyeongsan-siSouth Korea
  5. 5.Department of Chemical and Biomolecular EngineeringJohns Hopkins UniversityBaltimoreUSA
  6. 6.BrioBiotech LLCGlenelgUSA

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