Skip to main content

Advertisement

SpringerLink
Log in

Impact of nitrogen limitation on biomass, photosynthesis, and lipid accumulation in Chlorella sorokiniana

  • Published:
Journal of Applied Phycology Aims and scope Submit manuscript

Abstract

Induction of oil accumulation in algae for biofuel production is often achieved by withholding nitrogen. However, withholding nitrogen often reduces total biomass yield. In this report, it is demonstrated that Chlorella sorokiniana will not only accumulate substantial quantities of neutral lipids when grown in the absence of nitrogen but will also exhibit unimpeded growth rates for up to 2 weeks. To determine the physiological basis for the observed increase in oil and biomass accumulation, we compared photosynthetic and respiration rates and chlorophyll, lipid, and total energy content under ammonia replete and deplete conditions. Under N-depleted growth conditions, there was a 64 % increase in total energy density and a ∼20-fold increase in oil accumulation relative to N-replete growth leading to a 1.6-fold greater total energy yield in N-depleted than in N-replete cultures. We propose that the higher energy accumulation in N-depleted cultures is due to enhanced photosynthetic energy capture and conversion associated with reduced chlorophyll levels and reduced self-shading as well as a shift in metabolism leading to the accumulation of oils.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  • Berges JA, Charlebois DO, Mauzerall DC, Falkowski PG (1996) Differential effects of nitrogen limitation on photosynthetic efficiency of photosystems I and II in microalgae. Plant Physiol 110:689–696

    CAS  PubMed  PubMed Central  Google Scholar 

  • Breuer G, Lamers PP, Martens DE, Draaisma RB, Wijffels RH (2012) The impact of nitrogen starvation on the dynamics of triacylglycerol accumulation in nine microalgae strains. Bioresour Technol 124:217–226

    Article  CAS  PubMed  Google Scholar 

  • Cakmak T, Angun P, Demiray YE, Ozkan AD, Elibol Z, Tekinay T (2012) Differential effects of nitrogen and sulfur deprivation on growth and biodiesel feedstock production of Chlamydomonas reinhardtii. Biotechnol Bioeng 109:1947–1957

    Article  CAS  PubMed  Google Scholar 

  • Carvalho AP, Silva SO, Baptista JM, Malcata FX (2011) Light requirements in microalgal photobioreactors: an overview of biophotonic aspects. Appl Microbiol Biotechnol 89:1275–1288

    Article  CAS  PubMed  Google Scholar 

  • de Loura IC, Dubacq JP, Thomas JC (1987) The effects of nitrogen deficiency on pigments and lipids of cyanobacteria. Plant Physiol 83:838–843

    Article  PubMed  PubMed Central  Google Scholar 

  • Folch J, Lees M, Sloane Stanley GH (1957) A simple method for the isolation and purification of total lipides from animal tissues. J Biol Chem 226:497–509

    CAS  PubMed  Google Scholar 

  • Holguin F, Schaub T (2013) Characterization of microalgal lipid feedstocks by direct infusion FT-ICR mass spectrometry. Algal Res 2:43–50

    Article  Google Scholar 

  • Huerlimann R, de Nys R, Heimann K (2010) Growth, lipid content, productivity, and fatty acid composition of tropical microalgae for scale-up production. Biotechnol Bioeng 107:245–257

    Article  CAS  PubMed  Google Scholar 

  • Li T, Zheng Y, Yu L, Chen S (2013) High productivity cultivation of a heat-resistant microalga Chlorella sorokiniana for biofuel production. Bioresour Technol 131:60–67

    Article  CAS  PubMed  Google Scholar 

  • Lizzul AM, Hellier P, Purton S, Baganz F, Ladommatos N, Campos L (2014) Combined remediation and lipid production using Chlorella sorokiniana grown on wastewater and exhaust gases. Bioresour Technol 151:12–18

    Article  CAS  PubMed  Google Scholar 

  • Lourenco SO, Barbarino E (1998) Distribution of intrecellular nitrogen in marine microalgae: basis for the calculation of specific nitrogen-to-protein conversion factors. J Phycol 34:798–811

    Article  CAS  Google Scholar 

  • Lu C, Zhang J, Zhang Q, Li L, Kuang T (2001) Modification of photosystem II photochemistry in nitrogen deficient maize and wheat proteins. J Plant Physiol 158:1423–1430

    Article  CAS  Google Scholar 

  • Lv JM, Cheng LH, Xu XH, Zhang L, Chen HL (2010) Enhanced lipid production of Chlorella vulgaris by adjustment of cultivation conditions. Bioresour Technol 101:6797–6804

    Article  CAS  PubMed  Google Scholar 

  • Mackinney G (1941) Absorption of light by chlorophyll solutions. J Biol Chem 140:315–322

    CAS  Google Scholar 

  • Mata TM, Martins AA, Caetano NS (2010) Microalgae for biodiesel production and other applications: a review. Bioresour Technol 131:60–67

    Google Scholar 

  • Murphy TE, Berberoglu H (2012) Effect of algae pigmentation on photobioreactor productivity and scale-up: a light transfer perspective. J Quant Spectrosc Radiat Transf 112:2826–2834

    Article  Google Scholar 

  • Neidhardt J, Benemann JR, Zhang L, Melis A (1998) Photosystem-II repair and chloroplast recovery from irradiance stress: relationship between chronic photoinhibition, light-harvesting chlorophyll antenna size and photosynthetic productivity in Dunaliella salina (green algae). Photosynth Res 56:175–184

    Article  CAS  Google Scholar 

  • Perrine Z, Negi S, Sayre RT (2012) Optimization of photosynthetic light energy utilization by microalgae. Algal Res 1:134–142

    Article  Google Scholar 

  • Pruvost J, Van Vooren G, Le Gouic B, Couzinet-Mossion A, Legrand J (2011) Systematic investigation of biomass and lipid productivity by microalgae in photobioreactors for biodiesel application. Bioresour Technol 102:150–158

    Article  CAS  PubMed  Google Scholar 

  • Ramanna L, Guldhe A, Rawat I, Bux F (2014) The optimization of biomass and lipid yields of Chlorella sorokiniana when using wastewater supplemented with different nitrogen sources. Bioresour Technol 168:127–135

    Article  CAS  PubMed  Google Scholar 

  • Rosenberg JN, Kobayashi N, Barnes A, Noel EA, Betenbaugh MJ, Oyler GA (2014) Comparative analyses of three Chlorella species in response to light and sugar reveal distinctive lipid accumulation patterns in the microalga C. sorokiniana. PLoS One 9:e92460

    Article  PubMed  PubMed Central  Google Scholar 

  • Sayed OH, Hegazy AK (1992) Inhibition of secondary carotenoid biosynthesis during degreening of Chlorella fusca (Chlorococcales, Chlorophyta) and implication for growth and survival. Cryptogam Algol 13:181–186

    Google Scholar 

  • Sayre RT (2010) Microalgal biofuels; carbon capture and sequestration. Bioscience 60:722–727

    Article  Google Scholar 

  • Sharma KK, Schuhmann H, Schenk PM (2012) High lipid induction in microalgae for biodiesel production. Energies 5:1532–1553

    Article  CAS  Google Scholar 

  • Siaut M, Cuine S, Cagnon C, Fessler B, Nguyen M, Carrier P, Beyly A, Beisson F, Triantaphylides C, Li-Beisson Y, Peltier G (2011) Oil accumulation in the model green alga Chlamydomonas reinhardtii: characterization, variability between common laboratory strains and relationship with starch reserves. BMC Biotechnol 11:1–15

    Article  Google Scholar 

  • Simionato D, Block MA, La Rocca N, Jouhet J, Marechal E, Finazzi G, Morosinotto T (2013) Response of Nannochloropsis gaditana to nitrogen starvation includes a de novo biosynthesis of triacylglycerols, a decrease of chloroplast galactolipids and a reorganization of the photosynthetic apparatus. Eukaryot Cell 12:665–676

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Spolaore P, Joannis-Cassan C, Duran E, Isambert A (2006) Commercial applications of microalgae. J Biosci Bioeng 101:87–96

    Article  CAS  PubMed  Google Scholar 

  • Subramanian S, Barry AN, Pieris S, Sayre RT (2013) Comparative energetics and kinetics of autotrophic lipid and starch metabolism in chlorophytic microalgae: implications for biomass and biofuel production. Biotechnol Biofuels 6:150–162

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Sueoka N (1960) Mitotic replication of deoxyribonucleic acid in Chlamydomonas reinhardi. Proc Natl Acad Sci 46:83–91

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Tamburic B, Guruprasad S, Radford DT, Szabo M, Lilley RM, Larkum AW, Franklin JB, Kramer DM, Blackburn SI, Raven JA, Schliep M, Ralph PJ (2014) The effect of diel temperature and light cycles on the growth of Nannochloropsis oculata in a photobioreactor matrix. PLoS One 9:e86047

    Article  PubMed  PubMed Central  Google Scholar 

  • Tang H, Chen M, Garcia ME, Abunasser N, Ng KY, Salley SO (2011) Culture of microalgae Chlorella minutissima for biodiesel feedstock production. Biotechnol Bioeng 108:2280–2287

    Article  CAS  PubMed  Google Scholar 

  • Wang S-T, Pan Y-Y, Liu C-C, Chuang L-T, Chen C-NN (2011) Characterization of a green microalga UTEX 2219–4: effects of photosynthesis and osmotic stress on oil body formation. Bot Stud 52:305–312

    CAS  Google Scholar 

  • Zhang YM, Chen H, He CL, Wang Q (2013) Nitrogen starvation induced oxidative stress in an oil-producing green alga Chlorella sorokiniana C3. PLoS One 8:e69225

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgments

We thank Paige Pardington for her help in assisting with the photobioreactors during this experiment. This work is supported by the U.S. Department of Energy under contract DE-EE0003046 awarded to the National Alliance for Advanced Biofuels and Bioproducts for RTS and TS and by the Center for Animal Health and Food Safety at New Mexico State University for TS.

Author information

Authors and Affiliations

  1. New Mexico Consortium, Los Alamos, NM, 87544, USA

    Sangeeta Negi, Natalia Friedland, Sowmya Subramanian & Richard Sayre

  2. Bioscience Division, Los Alamos National Laboratory, Los Alamos, NM, 87545, USA

    Amanda N. Barry & Richard Sayre

  3. Center for Animal Health, Food Safety & Bio-Security, New Mexico State University, Las Cruces, NM, 88003, USA

    Nilusha Sudasinghe, F. Omar Holguin, Barry Dungan & Tanner Schaub

  4. Department of Biology, Missouri Baptist University, Saint Louis, MO, 63141, USA

    Shayani Pieris

Authors
  1. Sangeeta Negi
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Amanda N. Barry
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Natalia Friedland
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Nilusha Sudasinghe
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Sowmya Subramanian
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Shayani Pieris
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. F. Omar Holguin
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Barry Dungan
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Tanner Schaub
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Richard Sayre
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Richard Sayre.

Additional information

Sangeeta Negi and Amanda N. Barry shared first author

Electronic supplementary material

Below is the link to the electronic supplementary material.

ESM 1

(DOCX 436 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Negi, S., Barry, A.N., Friedland, N. et al. Impact of nitrogen limitation on biomass, photosynthesis, and lipid accumulation in Chlorella sorokiniana . J Appl Phycol 28, 803–812 (2016). https://doi.org/10.1007/s10811-015-0652-z

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10811-015-0652-z

Keywords

Search

Navigation