Journal of Applied Phycology

, Volume 30, Issue 4, pp 2325–2335 | Cite as

An efficient method for the sequential production of lipid and carotenoids from the Chlorella Growth Factor-extracted biomass of Chlorella vulgaris

  • Josephine Anthony
  • Kumar Thalavai Sivashankarasubbiah
  • Sreelatha Thonthula
  • Vijaya Raghavan Rangamaran
  • Dharani Gopal
  • Kirubagaran RamalingamEmail author


Efficient methodology for simultaneous extraction of multiple bioactive compounds from microalgae still remains a major challenge. The present study provides a method for the sequential production of three major products: Chlorella Growth Factor (CGF, a nucleotide-peptide complex enriched with vitamins, minerals, and carbohydrates), lipid, and carotenoids from Chlorella vulgaris biomass in an economically feasible manner. After protein-rich CGF was extracted, the spent biomass was found to contain 12% lipid and 3% carotenoids when extracted individually, compared to that of the un-utilized (fresh) biomass (lipid, 14%; carotenoids, 4%). When extracted simultaneously using conventional methods, the yield of lipid from “CGF and carotenoids-extracted biomass,” and carotenoids from “CGF and lipid-extracted biomass” were significantly reduced (50%). However, simultaneous extraction using different solvent mixtures such as hexane:methanol:water and pentane:methanol:water mixture-augmented lipid yield by 38.5% and carotenoids by 14%, and additionally retained chlorophyll and its derivatives. Column chromatographic approach yielded sequential production of lipid (18%), lutein (9%) with better yields as well as without chlorophyll interference. Different geometric isomers of lutein all-E-(trans)-(3R,3′R,6′R)-β,ε-carotene-3,3′diol, 9Z(cis)-(3R,3′R,6′R)-β,ε-carotene-3,3′diol, and 13Z(cis)-(3R,3′R,6′R)-β,ε-carotene-3,3′diol were purified by HPLC and elucidated by CD, UV, NMR, FT-IR, and Mass spectra. In conclusion, the study provides an efficient and economically viable methodology for sequential production of lipid and lutein along with its geometrical isomers without chlorophyll influence and yield loss from the protein-rich CGF-extracted spent biomass of marine microalga, Chlorella vulgaris.


Chlorella growth factor Lipid All-trans-lutein 9Z-lutein 13Z-lutein Chlorella vulgaris 



The authors gratefully acknowledge the Ministry of Earth Sciences, Govt. of India, for funding this project. The authors are thankful to Dr. S.S.C. Shenoi, Director, National Institute of Ocean Technology, for his constant support and encouragement. We are also grateful to the Council of Scientific and Industrial Research-Central Leather Research Institute (CSIR-CLRI), Chennai, for NMR, and Indian Institute of Technology (IIT), Chennai, for mass spectroscopic analyses. The Indian Institute of Chemical Technology (IICT), Hyderabad, and Indian Institute of Chromatography and Mass spectrometry (IICMS), Chennai, are also gratefully acknowledged for fatty acid and mass spectroscopic analyses, respectively. We would also like to thank Mr. Krupa Ratnam from the National Institute of Ocean Technology for his untiring help in presenting the figures.

Supplementary material

10811_2018_1430_MOESM1_ESM.doc (1 mb)
ESM 1 (DOC 1050 kb)


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Copyright information

© Springer Science+Business Media B.V., part of Springer Nature 2018

Authors and Affiliations

  • Josephine Anthony
    • 1
  • Kumar Thalavai Sivashankarasubbiah
    • 1
  • Sreelatha Thonthula
    • 1
  • Vijaya Raghavan Rangamaran
    • 1
  • Dharani Gopal
    • 1
  • Kirubagaran Ramalingam
    • 1
    Email author
  1. 1.Marine Biotechnology Division, National Institute of Ocean Technology, Ministry of Earth Sciences, Government of IndiaChennaiIndia

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