Skip to main content

Advertisement

SpringerLink
Log in

Simultaneous production of astaxanthin and lipids from Chlorella sorokiniana in the presence of reactive oxygen species: a biorefinery approach

  • Original Article
  • Published:
Biomass Conversion and Biorefinery Aims and scope Submit manuscript

Abstract

The current study aimed to investigate the concurrent production of astaxanthin and lipids using Chlorella sorokiniana under mixotrophic conditions in an external loop airlift photobioreactor (ELAPB). Supplementation of Fe2+ into the media in the red phase (stress phase) induced the astaxanthin production. The maximum yield of 154.36 mg L−1 was obtained by the end of the red phase, which is equivalent to 3.4% of the dry biomass weight. The role of reactive oxygen species in the formation of astaxanthin, which protects the microalgal cells from different oxidative stress, has been elucidated. Apart from astaxanthin, unsaturated fatty acids (81.34%) were also produced with excellent biodiesel properties. Unsaturated fatty acids viz., palmitoleic acid (16:1), have resulted in high yields (22.85%), followed by linoleic acid (18:2), 28.27%; oleic acid (C18:1), 14.38%; and eicosapentaenoic acid (20:5), 4.49%. Hence, the simultaneous production of high value-added products, viz., astaxanthin, and lipids makes the whole process economically viable and environmentally sustainable and elevates the commercial potential during scale-up. The astaxanthin radical scavenging activity was also assessed by H2O2 assay, and the maximum scavenging activity was determined as 91%. The study confers the potential advantages of algal cultivation for high-value commercial product synthesis towards additional revenue generation and development of an efficient microalgae-based biorefinery process.

Graphical abstract

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

  1. Ratnapuram HP, Vutukuru SS, Yadavalli R (2018) Mixotrophic transition induced lipid productivity in Chlorella pyrenoidosa under stress conditions for biodiesel production. Heliyon. 4:e00496. https://doi.org/10.1016/j.heliyon.2017.e00496

    Article  Google Scholar 

  2. Enamala MK, Dixit R, Tangellapally A, Singh M, Dinakarrao SMP, Chavali M, Pamanji SR, Ashokkumar V, Kadier A, Chandrasekhar K (2020) Photosynthetic microorganisms (algae) mediated bioelectricity generation in microbial fuel cell: concise review. Environ Technol Innov 19:100959. https://doi.org/10.1016/j.eti.2020.100959

    Article  Google Scholar 

  3. Aghajanpour M, Nazer MR, Obeidavi Z et al (2017) Functional foods and their role in cancer prevention and health promotion: a comprehensive review. Am J Cancer Res

  4. Hwang SW, Choi HIL, Sim SJ (2019) Acidic cultivation of Haematococcus pluvialis for improved astaxanthin production in the presence of a lethal fungus. Bioresour Technol 278:138–144. https://doi.org/10.1016/j.biortech.2019.01.080

    Article  Google Scholar 

  5. Lin WC, Chien JT, Chen BH (2005) Determination of carotenoids in spear shrimp shells (Parapenaeopsis hardwickii) by liquid chromatography. J Agric Food Chem 53:5144–5149. https://doi.org/10.1021/jf050664y

    Article  Google Scholar 

  6. Takaichi S, Matsui K, Nakamura M, Muramatsu M, Hanada S (2003) Fatty acids of astaxanthin esters in krill determined by mild mass spectrometry. Comp Biochem Physiol - B Biochem Mol Biol 136:317–322. https://doi.org/10.1016/S1096-4959(03)00209-4

    Article  Google Scholar 

  7. Sun Z, Li T, Zhou ZG, Jiang Y (2016) Microalgae as a source of lutein: chemistry, biosynthesis, and carotenogenesis. Adv Biochem Eng Biotechnol. https://doi.org/10.1007/10_2015_331

  8. Enamala MK, Enamala S, Chavali M, Donepudi J, Yadavalli R, Kolapalli B, Aradhyula TV, Velpuri J, Kuppam C (2018) Production of biofuels from microalgae - a review on cultivation, harvesting, lipid extraction, and numerous applications of microalgae. Renew Sust Energ Rev 94:49–68. https://doi.org/10.1016/j.rser.2018.05.012

    Article  Google Scholar 

  9. Zhang BY, Geng YH, Li ZK, Hu HJ, Li YG (2009) Production of astaxanthin from Haematococcus in open pond by two-stage growth one-step process. Aquaculture. 295:275–281. https://doi.org/10.1016/j.aquaculture.2009.06.043

    Article  Google Scholar 

  10. Zhang Y, Ye Y, Ding W, Mao X, Li Y, Gerken H, Liu J (2020) Astaxanthin is ketolated from zeaxanthin independent of fatty acid synthesis in Chromochloris zofingiensis. Plant Physiol 183:883–897. https://doi.org/10.1104/pp.20.00325

    Article  Google Scholar 

  11. Brotosudarmo THP, Limantara L, Setiyono E, Heriyanto (2020) Structures of astaxanthin and their consequences for therapeutic application. Int. J. Food Sci

  12. Wang B, Zarka A, Trebst A, Boussiba S (2003) Astaxanthin accumulation in Haematococcus pluvialis (Chlorophyceae) as an active photoprotective process under high irradiance. J Phycol 39:1116–1124. https://doi.org/10.1111/j.0022-3646.2003.03-043.x

    Article  Google Scholar 

  13. Kobayashi M (2003) Astaxanthin biosynthesis enhanced by reactive oxygen species in the green alga Haematococcus pluvialis. Biotechnol Bioprocess Eng 8:322–330

    Article  Google Scholar 

  14. Raman R, Mohamad SE (2012) Astaxanthin production by freshwater microalgae chlorella sorokiniana and marine microalgae Tetraselmis sp. Pakistan J Biol Sci 15:1182–1186. https://doi.org/10.3923/pjbs.2012.1182.1186

    Article  Google Scholar 

  15. Khalili Z, Jalili H, Noroozi M, Amrane A, Ashtiani FR (2020) Linoleic-acid-enhanced astaxanthin content of Chlorella sorokiniana (Chlorophyta) under normal and light shock conditions. Phycologia. 59:54–62. https://doi.org/10.1080/00318884.2019.1670012

    Article  Google Scholar 

  16. Poonkum W, Powtongsook S, Pavasant P (2015) Astaxanthin induction in microalga H. pluvialis with flat panel airlift photobioreactors under indoor and outdoor conditions. Prep Biochem Biotechnol. https://doi.org/10.1080/10826068.2013.867871

  17. Yadavalli R, S RR, Rao CS (2012) Lipid accumulation studies in Chlorella pyrenoidosa using customized photobioreactor- effect of nitrogen source, light intensity and mode of operation. Int J Eng Res Appl 2:2446–2453

    Google Scholar 

  18. Yin-Nin Ma R, Chen F (2001) Induction of astaxanthin formation by reactive oxygen species in mixotrophic culture of Chlorococcum sp. Biotechnol Lett 23:519–523. https://doi.org/10.1023/A:1010370401235

    Article  Google Scholar 

  19. Yadavalli R, Rao CS, Chandrakanth D, et al (2010) Effect of different culture media on cell concentrations of Chlorella pyrenoidosa under photoautotrophic conditions

  20. Inbaraj BS, Lu H, Hung CF, Wu WB, Lin CL, Chen BH (2008) Determination of carotenoids and their esters in fruits of Lycium barbarum Linnaeus by HPLC-DAD-APCI-MS. J Pharm Biomed Anal 47:812–818. https://doi.org/10.1016/j.jpba.2008.04.001

    Article  Google Scholar 

  21. Amorim-Carrilho KT, Cepeda A, Fente C, Regal P (2014) Review of methods for analysis of carotenoids. TrAC - Trends Anal Chem

  22. Yuan J, Fan L, Latasa P, et al (1999) Analysis of total astaxanthin in algae meal prepared from Haematococcus pluvialis. Aquasearch Inc 1:1–13

  23. Elumalai S, Santhose BI, Kanna GR (2014) Extraction of carotenoid and thin layer chromatography (TLC), GC-MS , FT-IR and HPLC analysis of pharmaceutically important pigment astaxanthin from a new strain of Haematococcus pluvialis. Wkly Sci Res J

  24. Yadavalli R, Ratnapuram H, Motamarry S, Reddy CN, Ashokkumar V, Kuppam C (2020) Simultaneous production of flavonoids and lipids from Chlorella vulgaris and Chlorella pyrenoidosa. Biomass Convers Biorefinery. https://doi.org/10.1007/s13399-020-01044-x

  25. BLIGH EG, DYER WJ (1959) A rapid method of total lipid extraction and purification. Can J Biochem Physiol 37:911–917. https://doi.org/10.1139/o59-099

    Article  Google Scholar 

  26. Yadavalli R, Ramgopal RS, Rao CS (2013) Lipid productivity of Chlorella pyrenoidosa in a customized lab scale photobioreactor under stress conditions. Int J ChemTech Res

  27. Lee JY, Yoo C, Jun SY, Ahn CY, Oh HM (2010) Comparison of several methods for effective lipid extraction from microalgae. Bioresour Technol 101:S75–S77. https://doi.org/10.1016/j.biortech.2009.03.058

    Article  Google Scholar 

  28. Yadavalli R, Peasari JR, Mamindla P, Praveenkumar, Mounika S, Ganugapati J (2018) Phytochemical screening and in silico studies of flavonoids from Chlorella pyrenoidosa. Informatics Med Unlocked 10:89–99. https://doi.org/10.1016/j.imu.2017.12.009

    Article  Google Scholar 

  29. Baviskar JW, Khandelwal SR (2015) Original research article extraction , detection and identification of flavonoids from microalgae: an emerging secondary metabolite. Int J Curr Microbiol Appl Sci

  30. Liu J, Sun Z, Gerken H, Liu Z, Jiang Y, Chen F (2014) Chlorella zofingiensis as an alternative microalgal producer of astaxanthin: biology and industrial potential. Mar Drugs 12:3487–3515

    Article  Google Scholar 

  31. Chai S, Shi J, Huang T, Guo Y, Wei J, Guo M, Li L, Dou S, Liu L, Liu G (2018) Characterization of chlorella sorokiniana growth properties in monosaccharide-supplemented batch culture. PLoS One 13:e0199873. https://doi.org/10.1371/journal.pone.0199873

    Article  Google Scholar 

  32. Li Y, Huang J, Sandmann G, Chen F (2009) High-light and sodium chloride stress differentially regulate the biosynthesis of astaxanthin in Chlorella zofingiensis (Chlorophyceae). J Phycol 45:635–641. https://doi.org/10.1111/j.1529-8817.2009.00689.x

    Article  Google Scholar 

  33. Ip PF, Chen F (2005) Employment of reactive oxygen species to enhance astaxanthin formation in Chlorella zofingiensis in heterotrophic culture. Process Biochem 40:3491–3496. https://doi.org/10.1016/j.procbio.2005.02.014

    Article  Google Scholar 

  34. Lam MK, Lee KT (2012) Microalgae biofuels: a critical review of issues, problems and the way forward. Biotechnol Adv 30:673–690

    Article  Google Scholar 

  35. Qiu B, Li Y (2006) Photosynthetic acclimation and photoprotective mechanism of Haematococcus pluvialis (Chlorophyceae) during the accumulation of secondary carotenoids at elevated irradiation. Phycologia. 45:117–126. https://doi.org/10.2216/04-99.1

    Article  Google Scholar 

  36. Han D, Li Y, Hu Q (2013) Astaxanthin in microalgae: pathways, functions and biotechnological implications. Algae

  37. Zhekisheva M, Boussiba S, Khozin-Goldberg I, Zarka A, Cohen Z (2002) Accumulation of oleic acid in Haematococcus pluvialis (Chlorophyceae) under nitrogen starvation or high light is correlated with that of astaxanthin esters. J Phycol 38:325–331. https://doi.org/10.1046/j.1529-8817.2002.01107.x

    Article  Google Scholar 

  38. Chang C Sen, Chang CL, Lai GH (2013) Reactive oxygen species scavenging activities in a chemiluminescence model and neuroprotection in rat pheochromocytoma cells by astaxanthin, beta-carotene, and canthaxanthin. Kaohsiung J Med Sci https://doi.org/10.1016/j.kjms.2012.12.002, 29, 412, 421

  39. Shim SJ, Hong ME, Chang WS, Sim SJ (2020) Repeated-batch production of omega-3 enriched biomass of Chlorella sorokiniana via calcium-induced homeoviscous adaptation. Bioresour Technol 303:122944. https://doi.org/10.1016/j.biortech.2020.122944

    Article  Google Scholar 

  40. Holtin K, Kuehnle M, Rehbein J, Schuler P, Nicholson G, Albert K (2009) Determination of astaxanthin and astaxanthin esters in the microalgae Haematococcus pluvialis by LC-(APCI)MS and characterization of predominant carotenoid isomers by NMR spectroscopy. Anal Bioanal Chem 395:1613–1622. https://doi.org/10.1007/s00216-009-2837-2

    Article  Google Scholar 

Download references

Acknowledgments

Y.R. and C.N.R. would like to thank the Management and Principal of CBIT for constant support and encouragement in carrying out this work.

Funding

The present work was funded by the Department of Biotechnology, Government of India, under the research grant # BT/PR13125/PBD/26/448/2015.

Author information

Authors and Affiliations

  1. Department of Biotechnology, Chaitanya Bharathi Institute of Technology (Autonomous), Gandipet, Hyderabad, Telangana State, 500075, India

    Rajasri Yadavalli & C. Nagendranatha Reddy

  2. Department of Biotechnology, Sreenidhi Institute of Science and Technology (Autonomous), Yamnampet, Ghatkesar, Hyderabad, Telangana State, India

    Rajasri Yadavalli, Hariprasad Ratnapuram & John Reddy Peasari

  3. Department of Chemical Technology, Chulalongkorn University, Bangkok, Thailand

    Veeramuthu Ashokkumar

  4. Green Processing, Bioremediation and Alternative Energies Research Group, Faculty of Environment and Labour Safety, Ton Duc Thang University, Ho Chi Minh City, Vietnam

    Chandrasekhar Kuppam

Authors
  1. Rajasri Yadavalli
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Hariprasad Ratnapuram
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. John Reddy Peasari
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. C. Nagendranatha Reddy
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Veeramuthu Ashokkumar
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Chandrasekhar Kuppam
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Chandrasekhar Kuppam.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Additional information

Publisher’s note

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

Highlights

• Chlorella sorokiniana is found as a potential microalgal species for the production of astaxanthin.

• The addition of ROS to the growth medium produced astaxanthin in the stress phase.

• Simultaneous lipid and astaxanthin production were favoured in the mixotrophic mode of cultivation, encouraging biorefinery concept when scaled up

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Yadavalli, R., Ratnapuram, H., Peasari, J.R. et al. Simultaneous production of astaxanthin and lipids from Chlorella sorokiniana in the presence of reactive oxygen species: a biorefinery approach. Biomass Conv. Bioref. 12, 881–889 (2022). https://doi.org/10.1007/s13399-021-01276-5

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s13399-021-01276-5

Keywords

Search

Navigation