Skip to main content
SpringerLink
Log in

Augmenting the expression of accD and rbcL genes using optimized iron concentration to achieve higher biomass and biodiesel in Chlorella vulgaris

  • Original Research Paper
  • Published:
Biotechnology Letters Aims and scope Submit manuscript

Abstract

Chlorella vulgaris is a form of microalgae commonly employed as a biological source of oil for biodiesel production. Major algal cultivation strategies are focused on stimulating growth rate and lipid content. In the present study, the algal growth media was supplemented with iron (III) chloride (FeCl3), as a stimulating factor for growth and lipid production, in three iron concentrations including 90, 200, and 500 µM. The turbidity of algal cells was measured on different days, to determine the growth rate. In optimum iron concentration, this measurement experienced a 2.1-fold increase. Next, the lipid content was extracted, and the amount of lipid produced in each treatment was calculated, which demonstrated a 4.57-fold increase in lipid productivity. The expression of genes corresponding to the metabolic enzymes (i.e. acetyl-CoA carboxylase (accD) and ribulose bisphosphate carboxylase large chain (rbcL)) was evaluated using real-time PCR under different initial iron feeds. As demonstrated in the results, the initial iron feed of 90 µM was an optimum concentration that obtained the highest growth rate, more cell density, and increased lipid production. In 90 µM initial iron concentration, the expression of accD and rbcL genes showed a 4.8- and 35-fold increase, respectively, compared to that of the control genes. Based on the results, this optimum iron concentration could satisfy the industrial interest in biodiesel production from C. vulgaris as a potential stimulating factor. However, higher levels of iron (e.g. 200 and 500 µM) failed to act as positive stress for increasing biodiesel production. Finally, in this paper, different mechanisms where iron affects acetyl-CoA carboxylase (ACCase) and 1,5-ribulose bisphosphate carboxylase/oxygenase (RuBisCo) are illustrated.

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
Fig. 6

Similar content being viewed by others

References

  • Baky AE, El-Baroty H, Bouaid S, Martinez A, Aracil M J (2012) Enhancement of lipid accumulation in Scenedesmus obliquus by Optimizing CO2 and Fe3+ levels for biodiesel production. Bioresour Technol 119:429–432

    Article  Google Scholar 

  • Andersson I, Backlund A (2008) Structure and function of Rubisco. Plant Physiol Biochem 46:275–291

    Article  CAS  Google Scholar 

  • Balk J, Schaedler TA (2014) Iron cofactor assembly in plants. Annu Rev Plant Biol 65:125–153

    Article  CAS  Google Scholar 

  • Bligh EG, Dyer WJ (1959) A rapid method of total lipid extraction and purification. Can J Biochem Physiol 37:911–917

    Article  CAS  Google Scholar 

  • Concas A, Steriti A, Pisu M, Cao G (2014) Comprehensive modeling and investigation of the effect of iron on the growth rate and lipid accumulation of C. vulgaris cultured in batch photobioreactors. Bioresource Technol 153:340–350

    Article  CAS  Google Scholar 

  • Dos Santos A, Kunigami C, Aranda D, Luz Lapa Teixeira C (2016) Assessment of triacylglycerol content in C.vulgaris cultivated in a two-stage process. Biomass Bioenerg 92:55–60

    Article  Google Scholar 

  • Estevez M, Malanga G, Puntarulo S (2001) Iron-dependent oxidative stress in Chlorella vulgaris. Plant Sci 161(1):9–17

    Article  CAS  Google Scholar 

  • Feng Y, Li C, Zhang D (2011) Lipid production of C. vulgaris cultured in artificial wastewater medium. Bioresour Technol 102(1):101–105

    Article  CAS  Google Scholar 

  • Fujii M, Rose AL, Omura T, Waite TD (2010) Effect of Fe (II) and Fe (III) transformation kinetics on iron acquisition by a toxic strain of Microcystis aeruginosa. Environ Sci Technol 44:1980–1986

    Article  CAS  Google Scholar 

  • Ge S, Champagne P, Plaxton WC, Leite GB, Marazzi F (2017) Microalgal cultivation with waste streams and metabolic constraints to triacylglycerides accumulation for biofuel production. Biofuels Bioprod Bioref 11:325–343

    Article  CAS  Google Scholar 

  • Gonçalves A, Pires J, Simões M (2013) Green fuel production: Processes applied to microalgae. Environ Chem Lett 11:315–324

    Article  Google Scholar 

  • Gong Y, Jiang M (2011) Biodiesel production with microalgae as feedstock: From strains to biodiesel. Biotechnol Lett 33:1269–1284

    Article  CAS  Google Scholar 

  • Ghosh S, Chandra N, Vishveshwara S (2015) Mechanism of iron-dependent repressor (IdeR) activation and dna binding: a molecular dynamics and protein structure network study. PLoS Comput Biol 11(12):1–24

    Article  CAS  Google Scholar 

  • Griffiths MJ, Harrison TL (2009) Lipid productivity as a key characteristic for choosing algal species for biodiesel production. J App Phycol 21(5):493–507

    Article  CAS  Google Scholar 

  • Hamedi B, Mahdavi MA, Gheshlaghi R (2016) Improved lipid and biomass productivities in Chlorella vulgaris by differing the inoculation medium from the production medium. Biofuel Res J 10:410–416

    Article  Google Scholar 

  • Hantke K (2002) Members of the fur protein family regulate iron and zinc transport in E. coli and characteristics of the fur-regulated fhuf protein. J Mol Microbiol Biotechnol 4:217–222

    CAS  PubMed  Google Scholar 

  • Henry S, Gaspar M, Jesch S (2014) The response to inositol: regulation of glycerolipid metabolism and stress response signaling in yeast. Chem Phys Lipids 180:23–43

    Article  CAS  Google Scholar 

  • Huerlimann AR, Zenger KR, Jerry DR, Heimann K (2015) Phylogenetic analysis of nucleus-encodedacetyl-coa carboxylases targeted at thecytosol and plastid of algae. PLoS One 10(7):e0131099

    Article  Google Scholar 

  • Kumar R, Biswas K, Singh PK, Elumalai S (2017) Lipid production and molecular dynamics simulation for regulation of accD gene in cyanobacteria under different N and P regimes. Biotechnol Biofuels 10(1):94

    Article  Google Scholar 

  • Lewandowska J, Kosakowska A (2004) Effect of iron limitation on cells of the diatom Cyclotella meneghiniana Kützing. Oceanologia 46(2):269–287

    Google Scholar 

  • Li YX, Zhao FG, Yu DD (2015) Effect of nitrogen limitation on cell growth, lipid accumulation and gene expression in Chlorella sorokiniana. Braz arch biol technol 58:462–467

    Article  CAS  Google Scholar 

  • Li H, Chen W, Jin R, Jin JM, Tang SY (2017) Biosensor-aided high-throughput screening of hyper-producing cells for malonyl-CoA-derived products. Microb Cell Fact 16:187

    Article  Google Scholar 

  • Liu ZY, Wang GC, Zhou BC (2008) Effect of iron on growth and lipid accumulation in Chlorella vulgaris. Bioresour Technol 99:4717–4722

    Article  CAS  Google Scholar 

  • Luo S, Ishida H, Makino A, Mae T (2002) Fe2+-catalyzed Site-specific cleavage of the large subunit of ribulose 1,5-bisphosphate carboxylase close to the active site. J Biol Chem 277:12382–12387

    Article  CAS  Google Scholar 

  • Madadi R, Tabatabaei M, Aghbashlo M, Zahed MA, Pourbabaee AA (2018) Biodiesel from microalgae. Waste to Wealth. Editors: Singhania, R.R., Agarwal, R.A., Kumar, R.P., Sukumaran, R.K 277–318. Springer, New York

  • Mata T, Martins A, Caetano N (2009) Microalgae for biodiesel production and other applications: A review. Renew Sust Energ Rev 14:217–232

    Article  Google Scholar 

  • Manuelle N, Courchesne D, Parisien A, Wang B, Lan C (2009) Enhancement of lipid production using biochemical, genetic and transcription factor engineering approaches. J Exp Bot 66:3725–3735

    Google Scholar 

  • Menon KR, Balan R, Suraishkumar G (2013) Stress induced lipid production in Chlorella vulgaris: Relationship with specific intracellular reactive species levels. Biotechnol Bioeng 110:1627–1636

    Article  CAS  Google Scholar 

  • Plank DW, Gengenbach BG, Gronwald JW (2001) Effect of iron on activity of soybean multi-subunit acetyl-coenzyme A carboxylase. Physiol plantarum 112:183–194

    Article  CAS  Google Scholar 

  • Ren HY, Liu BF, Kong F, Zhao L, Xie GJ, Ren NQ (2014) Enhanced lipid accumulation of green microalga Scenedesmus sp. by metal ions and EDTA addition. Bioresour Technol 169:763–767

    Article  CAS  Google Scholar 

  • Saad MG, Dosoky NS, Zoromba MS, Shafik HM (2019) Algal Biofuels: Current Status and Key Challenges. Energies 12: 1920

  • Sato J, Takeda K, Nishiyama R, Fusayama K, Arai T, Sato T, Watanabe T, Abe A, Nakagawa J, Kawasaki S, Nimura Y (2010) Chlorella vulgaris aldehyde reductase is capable of functioning as ferric reductase and of driving the fenton reaction in the presence of free flavin. Biosci Biotechnol Biochem 74(4):854–857

    Article  CAS  Google Scholar 

  • Singh P, Kumari S, Guldhe A, Misra R, Rawat I, Bux F (2016) Trends and novel strategies for enhancing lipid accumulation and quality in microalgae. Renew Sust Energ Rev 55:1–16

    Article  CAS  Google Scholar 

  • Singh P, Kumari S, Guldhe A, Singh G, Bux F (2017) Accase and rbcL gene expression as a function of nutrient and metal stress for enhancing lipid productivity in Chlorella sorokiniana. Energy Convers Manag 148:809–819

    Article  CAS  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 

  • Stephenson A, Dennis J, Howe C, Scott S, Smith A (2010) Influence of nitrogen limitation regime on the production by C.vulgaris of lipids for biodiesel feedstocks. Biofuels 1:47–58

    Article  CAS  Google Scholar 

  • Sutak R, Botebol H, Blaiseau PL, Léger T, Bouget FY, Camadro JM, Lesuisse E (2012) A comparative study of iron uptake mechanisms in marine microalgae: iron binding at the cell surface is a critical step1[W][OA]. Plant Physiol 160:2271–2284

    Article  CAS  Google Scholar 

  • Talebi AF, Mohtashami SK, Tabatabaei M, Tohidfar M, Bagheri A, Zeinalabedini M, Hadavand Mirzaei H, Mirzajanzadeh M, Malekzadeh Shafaroudi S, Bakhtiari S (2013) Fatty acids profiling: a selective criterion for screening microalgae strains for biodiesel production. Algal Res 2:258–267

    Article  Google Scholar 

  • Talebi AF, Tohidfar M, Bagheri A, Lyon S, Salehi Ashtiani K, Tabatabaei M (2014) Manipulation of carbon flux into fatty acid biosynthesis pathway in Dunaliella salina using accD and ME genes to enhance lipid content and to improve produced biodiesel quality. Biofuel Res J 3:91–97

    Article  Google Scholar 

  • Talebi AF, Tabatabaei M, Aghbashlo M (2018) Recent patents on biofuels from microalgae. In: Jacob-Lopes E et al (eds) Energy from microalgae, green energy and technology. Springer, New York, pp 291–306

  • Terauchi AM, Peers G, Kobayashi MC, Niyogi KK, Merchant SS (2010) Trophic status of Chlamydomonas reinhardtii influences the impact of iron deficiency on photosynthesis. Photosyn Res 105:39–49

    Article  CAS  Google Scholar 

  • Wan M, Jin X, Xia J, Rosenberg JN, Yu G, Nie Z, Oyler GA, Betenbaugh MJ (2014) The effect of Iron on growth, lipid accumulation, and gene expression profile of the freshwater microalga Chlorella sorokiniana. Appl Microbiol Biotechnol 98:9473–9481

    Article  CAS  Google Scholar 

  • Zhang J, Hao Q, Bai L, Xu J, Yin W, Song L, Xu L, Guo X, Fan C, Chen Y, Ruan J, Hao S, Li Y, Wang R, Hu Z (2014) Overexpression of the soybean transcription factor GmDof4 significantly enhances the lipid content of Chlorella ellipsoidea. Biotechnol Biofuels 7:128

    PubMed  PubMed Central  Google Scholar 

  • Zhu LD, Li ZH, Hiltunen E (2016) Strategies for lipid production improvement in microalgae as a biodiesel feedstock. Biomed Res Int 1–8

Download references

Acknowledgements

We would like to thank Mr. Kaveh Khosraviani, University of Tehran, College of agriculture and natural resources, Dr. Saeed Iriana, Kharazmi University, department of the Cell and Molecular Biology, Faculty of biological sciences, Tehran, Iran, for their contribution in molecular sections.

Author information

Authors and Affiliations

  1. Department of Cell and Molecular Biology, Faculty of Biological Sciences, Kharazmi University, Tehran, Iran

    Atefeh Khamoushi, Vida Tafakori, Mohammad Ali Zahed, Somayeh Eghrari Gayglou & S. Abdolhamid Angaji

Authors
  1. Atefeh Khamoushi
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Vida Tafakori
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Mohammad Ali Zahed
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Somayeh Eghrari Gayglou
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. S. Abdolhamid Angaji
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Vida Tafakori.

Ethics declarations

Conflict of interest

There is no conflict of interest to declare.

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Khamoushi, A., Tafakori, V., Zahed, M.A. et al. Augmenting the expression of accD and rbcL genes using optimized iron concentration to achieve higher biomass and biodiesel in Chlorella vulgaris. Biotechnol Lett 42, 2631–2641 (2020). https://doi.org/10.1007/s10529-020-02973-3

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10529-020-02973-3

Keywords

Search

Navigation