Journal of Food Science and Technology

, Volume 51, Issue 2, pp 322–328 | Cite as

Enzymatic and non-enzymatic antioxidant potentials of Chlorella vulgaris grown in effluent of a confectionery industry

  • R. Ranjith Kumar
  • P. Hanumantha Rao
  • V. V. Subramanian
  • V. Sivasubramanian
Original Article


Enzymatic and non-enzymatic antioxidant potentials of Chlorella vulgaris have gained considerable importance in recent decades. C. vulgaris strain highly tolerant to extreme pH variations was isolated and mass-cultivated in the wastewater from a confectionery industry. C.vulgaris showed better growth in wastewater than in improvised CFTRI medium. The microalgal biomass was then screened for the following antioxidants: peroxidase, superoxide dismutase, polyphenol oxidase, glutathione peroxidase, chlorophyll a, ascorbic acid, α-tocopherol and reduced glutathione. The total polyphenol content of the strain was also studied. The strain showed a high degree of enzymatic antioxidant activity (0.195 × 10−5 ± 0.0072 units/cell peroxidase, 0.04125 × 10−5 ± 0.001 units/cell superoxide dismutase, 0.2625 × 10−5 ± 0.003 units/cell polyphenol oxidase and 0.025 × 10−5 ± 0.003 glutathione peroxidase). The microalgal biomass also showed, per milligram weight, 0.2182 ± 0.005 μg of ascorbic acid, 0.00264 ± 0.001 μg of α-tocopherol and 0.07916 ± 0.004 μg of reduced glutathione. These results represent the possibility of using C. vulgaris grown in confectionery industry wastewater as a source of nutritious supplement, which is highly promising in terms of both economic and nutritional point of view.


Chlorella vulgaris Confectionery industry effluent Enzymatic antioxidants Non-enzymatic antioxidants 


  1. Anbuselvam C, Vijayavel K, Balasubramanian MP (2007) Protective effect of Operculina turpethum against 7,12-dimethyl benz(a)anthracene induced oxidative stress with reference to breast cancer in experimental rats. Chem Biol Interact 168:229–236CrossRefGoogle Scholar
  2. Choudhary R, Saroha AE, Swarnkar PL (2011) Effect of abscisic acid and hydrogen peroxide on antioxidant enzymes in syzygium cumini plant. J Food Sci Tech. doi: 10.1007/s13197-011-0464-3
  3. Becker EW (1994) Microalgae. In: Baddiley SJ, Carey NH, Higgins IJ, Potter WG (eds) Biotechnology-Microbiology. Cambridge University Press, New York, pp 177–195Google Scholar
  4. Bligh EG, Dyer WJ (1959) A rapid method for total lipid extraction and purification. Can J Biochem Physiol 37:911–917CrossRefGoogle Scholar
  5. Borowitzka M (1995) Microalgae as sources of pharmaceuticals and other biologically active compounds. J Appl Phycol 7:3–15CrossRefGoogle Scholar
  6. Borowitzka MA (1999a) Commercial production of microalgae: ponds, tanks, tubes and fermenters. J Biotechnol 70:313–321CrossRefGoogle Scholar
  7. Borowitzka MA (1999b) Economic evaluation of microalgal processes and products. In: Cohen Z (ed) Chemicals from microalgae. Taylor & Francis, London, pp 387–409Google Scholar
  8. Bo Shao H, Ye Chu L, Hua Lu Z, Min Kang C (2008) Primary antioxidant free radical scavenging and redox signaling pathways in higher plant cells. Int J Biol Sci 4:8–14Google Scholar
  9. Burguieres E, McCue P, Kwon YI, Shetty K (2007) Effect of vitamin C and folic acid on seed vigor response and phenolic-linked antioxidant activity. Bioresour Technol 98:1393–1404CrossRefGoogle Scholar
  10. Cornet JF, (1998) Le technoscope: les photobioréacteurs. Biofutur 176:1–10Google Scholar
  11. de-Bashan LE, Moreno M, Hernandez JP, Bashan Y (2002) Removal of ammonium and phosphorus ions from synthetic wastewater by the microalgae Chlorella vulgaris coimmobilized in alginate beads with the microalgae growth-promoting bacterium Azospirillum brasilense. Water Res 36:2941–2948CrossRefGoogle Scholar
  12. Esterbauer H, Schwarzl E, Hayn M (1977) A rapid assay for catechol oxidase and laccase using 2-nitro-5-thiobenzoic acid. Anal Biochem 77:486–494CrossRefGoogle Scholar
  13. Garcia O, De-bashan EL, Hernandez J, Bashan Y (2010) Efficiency of growth and nutrient uptake from wastewater by heterotrophic, autotrophic, and mixotrophic cultivation of Chlorella vulgaris immobilized with azospirillum brasilense. J Appl Phycol 46:800–812CrossRefGoogle Scholar
  14. Gitta SJ, Donald I, Ginsberg MS, Christian Drapeau MS (2001) Blue-green algae as an immuno-enhancer and biomodulator. JANA 3:24–30Google Scholar
  15. Guil-Guerrero JL, Navarro-Juárez R, López-Martínez JC, Campra-Madrid P, Rebolloso-Fuentes MM (2004) Functionnal properties of the biomass of three microalgal species. J Agr Food Chem 65:511–517Google Scholar
  16. Han XU, Miao X, Qingyu W (2006) High quality biodisel production from a microalgae chlorella protothecoides by hetrotrophic growth in fermenters. J Biotechnol 126:499–507CrossRefGoogle Scholar
  17. Hanumantha Rao P, Ranjith Kumar R, Raghavan BG, Subramanian VV, Sivasubramanian V (2011) Application of phycoremediation technology in the treatment of wastewater from a leather-processing chemical manufacturing facility. Water SA 37:7–14Google Scholar
  18. Herrero M, Jaime L, Martín-Álvarez PJ, Cifuentes A, Ibañez E (2006) Optimization of the extraction of antioxidants from Dunaliella salina microalga by pressurized liquids. J Agr Food Chem 54:5597–5603CrossRefGoogle Scholar
  19. Jaime L, Mendiola J, Herrero M, Soler-Rivas C, Santoyo S, Senorans F, Cifuentes A, Ibanez E (2005) Separation and characterization of antioxidants from Spirulinaplatensis microalga combining pressurized liquid extraction, TLC and HPLCDAD. J Sep Sci 28:2111–2119CrossRefGoogle Scholar
  20. Jeffrey SW, Humphrey GF (1975) New spectrophotometric equations for determining chlorophyll a, b, c1 and c2 in higher plants, algae and natural phytoplankton. Biochem Physiol Pflanz 167:191–194Google Scholar
  21. Karpinski S, Reynolds H, Karpinksa B, Wingsle G, Creissen G, Mullineaux P (1999) Systemic signaling and acclimation in response to excess excitation energy in Arabidopsis. Science 284:654–657CrossRefGoogle Scholar
  22. Komor E, Tanner W (1976) Glucose induces two amino acid transport systems in Chlorella. Eur J Biochem 70:197–204CrossRefGoogle Scholar
  23. Lamoureux G, Rusness D (1989) The role of glutathione and glutathione S-transferase in pesticide metabolism, selectivity, and mode of action in plants and insects. In: Poulson R, Avramovic O, Dolphin D (eds) Coenzymes and cofactors glutathione: chemical, biochemical, and medical aspects, part B. John Wiley and Sons, New York, pp 153–196Google Scholar
  24. Lei A, Hu Z, Wang Y, Tam NF (2006) Antioxidant responses of microalgae species to pyrene. J Appl Phycol 18:67–78 Google Scholar
  25. Lewis N, Yamamoto E (1990) Lignin: Occurrence, biogenesis and biodegradation. Annu Rev Plant Physiol Plant Mol Biol 41:455–496CrossRefGoogle Scholar
  26. Li H, Cheng K, Wong C, Fan K, Chen F, Jiang Y (2007) Evaluation of antioxidant capacity and total phenolic content of different fractions of selected microalgae. Food Chem 102:771–776CrossRefGoogle Scholar
  27. Li M, Hu C, Zhu Q (2006) Copper and zinc induction of lipid peroxidation and effects on antioxidant enzyme activities in the microalga Pavlova viridis (Prymnesiophyceae). Chemosphere 62:565–572CrossRefGoogle Scholar
  28. Lowry LH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurements with the folin-phenol reagent. J Biol Chem 193:265–275Google Scholar
  29. Malik CP (1980) In: Singh MB (ed) Plant enzymology and histo-enzymology. Kalyani Publishers, New Delhi, pp 69–80Google Scholar
  30. Matsukawa R, Hotta M, Masuda Y, Chihara M, Karube I (2000) Antioxidants from carbon dioxide fixing Chlorella sorokiniana. J Appl Phycol 12:263–267CrossRefGoogle Scholar
  31. Misra HP, Fridovich I (1972) The role of superoxide anion in the autoxidation of epinephrine and a simple assay for superoxide dismutase. J Biol Chem 247:3170–3175Google Scholar
  32. Moron M, Depierre J, Mannervik B (1979) Levels of glutathione, glutathione reductase and glutathione S-transferase activities in rat lung and liver. Biochim Biophys Acta 582:67–78CrossRefGoogle Scholar
  33. Munoz R, Guieysee B (2006) Algal-bactrial processes for the treatment of hazardous contaminants: A review. Water Res 40:2799–2815CrossRefGoogle Scholar
  34. Nocto G, Foyer C (1988) Ascorbate and glutathione: keeping active oxygen under control. Ann Rev Plant Physiol Plant Mol Biol 49:249–279CrossRefGoogle Scholar
  35. Nasirullah JT, Rakshitha D (2009) Isolation and antioxidant efficacy of nutraceutical concentrates from sesame and flax seed oils. J Food Sci Tech 46:66–69Google Scholar
  36. Philipose MT, ICAR Monograph (1967) Chlorococcales. Algae, New Delhi, p 365Google Scholar
  37. Pons A, Rola P, Agvilo C, Garcia FJ, Alemarry M, Paloo A (1981) A method for the simultaneous determination of total carbohydrate and glycerol in biological samples with the anthrone reagent. J Biochem Biophys Methods 4:227–231CrossRefGoogle Scholar
  38. Rao AR, Sarada R, Baskaran V, Ravishankar GA (2006) Antioxidant activity of Botryococcus braunii extract elucidated in vitro models. J Agr Food Chem 54:4593–4599CrossRefGoogle Scholar
  39. Reddy K, Subhani S, Khan P, Kumar K (1985) Effect of light and benzyladenine on dark treated growing rice (Oryza sativa) leaves II. Changes in peroxidase activity. Plant Cell Physiol 26:987–994Google Scholar
  40. Rodriguez-Garcia I, Guil-Guerrero JL (2008) Evaluation of the antioxidant activity of three microalgal species for use as dietary supplements and in the preservation of foods. Food Chem 108:1023–1026CrossRefGoogle Scholar
  41. Roe JH, Kuether CA (1943) The determination of ascorbic acid in whole blood and urine through 2, 4 - dinitrophenyl hydrazine derivative of Dehydroascorbic acid. J Biol Chem 147:399–407Google Scholar
  42. Rosenberg HR (1992) Chemistry and physiology of the vitamins. Interscience Publishers, New York, pp 452–453Google Scholar
  43. Rotruk JT, Pope AL, Ganther HE, Swanson AB, Hafeman DG, Hoeckstra WG (1973) Selenium: Biochemical role as a component of glutathione peroxidase. Science 179:588–590CrossRefGoogle Scholar
  44. Roy G, Sarma BK, Phadnis PP, Mugesh G (2005) Selenium-containing enzymes in mammals: Chemical perspectives. J Chem Sci 117:287–303CrossRefGoogle Scholar
  45. Sachindra NM, Airanthi MKWA, Hosokawa M, Miyashita K (2010) Radical scavenging and singlet oxygen quenching activity of extracts from Indian seaweeds. J Food Sci Tech 47(1):94–99CrossRefGoogle Scholar
  46. Santos I, Almeida J, Salema R (1999) The influence of UV-B radiation on the superoxide dismutase of maize potato sorghum and wheat leaves. Can J Bot 77:70–76Google Scholar
  47. Singh UAK, Singh S, Rai M (2009) Total phenolics content and free radical scavenging activity of brassica vegetables. J Food Sci Tech 46:595–597Google Scholar
  48. Spolaore P, Joannis-Cassan C, Duran E, Isambert A (2006) Commercial applications of microalgae. J Biosci Bioeng 101:87–96CrossRefGoogle Scholar
  49. Takekoshi H, Suzuki G, Chubachi H, Nakano M (2005) Effect of Chlorella pyrenoidosa on facel extraction and liver accumulation of polychlorinated dibenzo-dioxin in mice. Chemosphere 59:297–304CrossRefGoogle Scholar
  50. Tsavalos AJ, Day JG (1994) Development of media for the mixotrophic/heterotrophic culture of Brachiomonas submarina. J Appl Phycol 6:431–433CrossRefGoogle Scholar
  51. Venkataraman LV, Becker EW (1985) Biotechnology & utilization of algae - The Indian experience. Sharada Press, Mangalore, pp 114–115Google Scholar
  52. Wu L, Ho JAA, Shieh M, Lu I (2005) Antioxidant and antiproliferative activities of Spirulina and Chlorella water extracts. J Agr Food Chem 53:4207–4212CrossRefGoogle Scholar
  53. Yang C, Ding Z, Zhang K (2008) Growth of Chlorella pyrenoidosa in wastewater from cassava ethanol fermentation. World J Microbiol Biotechnol 24:2919–2925CrossRefGoogle Scholar

Copyright information

© Association of Food Scientists & Technologists (India) 2011

Authors and Affiliations

  • R. Ranjith Kumar
    • 1
  • P. Hanumantha Rao
    • 1
  • V. V. Subramanian
    • 1
  • V. Sivasubramanian
    • 1
  1. 1.Department of Plant Biology and Plant BiotechnologyRamakrishna Mission Vivekananda CollegeChennaiIndia

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