ISOLATION AND CHARACTERIZATION OF ENDOPHYTIC FUNGI FROM CALOTROPIS PROCERA FOR THEIR ANTIOXIDANT ACTIVITY
Objective: To investigate the antioxidant activity of methanolic extract of endophytic fungi isolated from Calotropis procera plant.
Methods: The endophytic fungi were isolated from explants of leaf, stem, and flower of C. procera on potato dextrose broth (PDB) medium. The fungal
isolates were mass cultured in PDB. The methanolic extracts of these fungi were prepared and evaluated for the antioxidant activity by 1,1-diphenyl-
2-picryl-hydrazyl (DPPH) free radical scavenging assay and reducing power assay. The extracts were characterized for the presence of phenolics and
Results: A total of 8 endophytic fungi were isolated from C. procera explants. Out of these 8 fungal isolates, the crude extracts of two fungi, i.e. CPIMR-2
and CPIL-1 showed positive antioxidant activity. From the morphological characteristics, the isolates CPIMR-2 and CPIL-1 were identified as Penicillium
spp. and Aspergillus spp., respectively. Both the isolates were found to possess antioxidant potential with % inhibition value of 81.16% and 80.97%
in the DPPH radical scavenging assay. The phytochemical screening of the methanolic extracts showed the presence of phenolics and flavonoids. The
total flavonoids content in Penicillium species and Aspergillus species were found to be 130.50 Î¼g/mg and 94.91 Î¼g/mg of extracts, respectively. The
total phenolic content was found to be 9.16 Î¼g/mg and 12.13 Î¼g/mg of extracts in Penicillium species and Aspergillus species, respectively.
Conclusions: Endophtyic fungi were found to be present in medicinal value plant C. procera. The probable bioactive component for antioxidant
activity possessed by the fungi would be the flavonoids and phenolics. These metabolite produced by endophytic fungi from C. procera need to be
explored further as potential source of novel natural antioxidant compound.
Keywords: Endophytes, Calotropis procera, Antioxidant, Radical scavenging, Reducing power, Flavonoids.
1. Halliwell B. Free radicals and antioxidants: A personal view. Nutr Rev 1994;52:253-65.
2. Finkel T, Holbrook NJ. Oxidants, oxidative stress and the biology of ageing. Nature 2000;408(6809):239-47.
3. KÃ¤hkÃ¶nen MP, Hopia AI, Vuorela HJ, Rauha JP, Pihlaja K, Kujala TS, et al. Antioxidant activity of plant extracts containing phenolic compounds. J Agric Food Chem 1999;47(10):3954-62.
4. Zheng W, Wang SY. Antioxidant activity and phenolic compounds in selected herbs. J Agric Food Chem 2001;49(11):5165-70.
5. Cai Y, Luo Q, Sun M, Corke H. Antioxidant activity and phenolic compounds of 112 traditional Chinese medicinal plants associated with anticancer. Life Sci 2004;74(17):2157-84.
6. BÃ©rdy J. Bioactive microbial metabolites. J Antibiot (Tokyo) 2005;58(1):1-26.
7. Tan RX, Zou WX. Endophytes: A rich source of functional metabolites. Nat Prod Rep 2001;18(4):448-59.
8. Yu H, Zhang L, Li L, Zheng C, Guo L, Li W, et al. Recent developments and future prospects of antimicrobial metabolites produced by endophytes. Microbiol Res 2010;165(6):437-49.
9. Harper JK, Arif AM, Ford EJ, Strobel GA, Porco JA, Tomer DP. Pestacin a 1, 3-dihydro isobenzofuran from Pestalotiopsis microspora possessing antioxidant and antimycotic activities. Tetrahedron 2003;59(14):2471-6.
10. Huang XZ, Zhu Y, Guan XL, Tian K, Guo JM, Wang HB, et al. A novel antioxidant isobenzofuranone derivative from fungus Cephalosporium sp.AL031. Molecules 2012;17(4):4219-24.
11. Song YC, Huang WY, Sun C, Wang FW, Tan RX. Characterization of graphislactone A as the antioxidant and free radical-scavenging substance from the culture of Cephalosporium sp. IFB-E001, an endophytic fungus in Trachelospermum jasminoides. Biol Pharm Bull 2005;28(3):506-9.
12. Zhao J, Fu Y, Luo M, Zu Y, Wang W, Zhao C, et al. Endophytic fungi from pigeon pea [Cajanus cajan (L.) Millsp.] produce antioxidant Cajaninstilbene acid. J Agric Food Chem 2012;60(17):4314-9.
13. Huang WY, Cai YZ, Xing J, Corke H, Sun M. A potential antioxidant resource: Endophytic fungi from medicinal plants. Econ Bot 2007;61:14-30.
14. Liu X, Dong M, Chen X, Jiang M, Lv X, Yan G. Antioxidant activity and phenolics of an endophytic Xylaria sp. from Ginkgo biloba. Food Chem 2007;105(2):548-54.
15. Schulz B, Wanke U, Draeger S, Aust HJ. Endophytes from herbaceous plants and shrubs: Effectiveness of surface sterilization methods. Mycol Res 1993;97(12):1447-50.
16. Goveas SW, Madtha R, Nivas SK, Dâ€™Souza L. Isolation of endophytic fungi from Coscinium fenestratum-a red listed endangered medicinal plant. Euras J Bio Sci 2011;5(4):48-53.
17. Lin Y, Wang J, Wu X, Zhou S, Vrijmoed LL, Jones EG. A novel compound enniatin G from the mangrove fungus Halosarpheia sp.(strain# 732) from the South China sea. Aust J Chem 2002;55(3):225-7.
18. Choudhary MI, Musharraf SG, Mukhmoor T, Shaheen F, Ali S, Rahman AU. Isolation of bioactive compounds from Aspergillus terreus. Z Naturforsch B 2004;59:324-8.
19. Babu DR, Rao GN. Antioxidant properties and electrochemical behavior of cultivated commercial Indian edible mushrooms. J Food Sci Technol 2013;50(2):301-8.
20. Oyaizu M. Studies on products of browning reaction--antioxidative activities of products of browning reaction prepared from glucosamine. Jpn J Nutr 1986;44(6):307-15.
21. Trease GE, Evans WC. Pharmacognosy. 13th ed. London: ELBS, Bailliere Tindall; 1989. p. 345-6.
22. Wagner H, Bladt S. Plant Drug Analysis: A Thin Layer Chromatography atlas. Heidelberg: Springer Science & Business Media; 1996.
23. Sathishkumar T, Baskar R, Shanmugam S, Rajasekaran P, Sadasivam S, Manikandan V. Optimization of flavonoids extraction from the leaves of Tabernaemontana heyneana Wall. using L16 Orthogonal design. Nat Sci 2008;6(3):10-21.
24. Patel VR, Patel PR, Kajal SS. Antioxidant activity of some selected medicinal plants in western region of India. Adv Biol Res 2010;4(1):23-6.
25. Singleton VL, Orthofer R, Lamuela-Raventos RM. Analysis of total phenols and other oxidation substrates and antioxidants by means of folin-ciocalteu reagent. Method Enzymol 1999;299:152-78.
26. Khan R, Shahzad S, Choudhary MI, Khan SA, Ahmad A. Biodiversity of the endophytic fungi isolated from Calotropis procera (Ait.) R. Br. Pak J Bot 2007;39(6):2233-9.
27. Selvanathan S, Indrakumar I, Johnpaul M. Biodiversity of the endophytic fungi isolated from Calotropis gigantea (L.) R. Br. Rec Res Sci Technol 2011;3(4):94-100.
28. Kharwar RN, Verma VC, Strobel G, Ezra D. The endophytic fungal complex of Catharanthus roseus (L.) G. Don. Curr Sci 2008;95:228-33.
29. Dhankhar S, Kumar S, Dhankhar S, Yadav JP. Antioxidant activity of fungal endophytes isolated from Salvadora oleoides Decne. Int J Pharm Pharm Sci 2012;4(2):380-5.
30. Kalita P, Tapan BK, Pal TK, Kalita R. Estimation of total flavonoids content (TFC) and anti oxidant activities of methanolic whole plant extract of Biophytum sensitivum Linn. J Drug Deliv Ther 2013;3(4):33-7.
31. Abirami G, Boominathan M. Antimicrobial activity of endophytic fungi isolated from medicinal plant Hugonia mystax L. J Acad Ind Res 2016;4(12):257-62.
32. Ruma K, Kumar S, Prakash HS. Antioxidant, anti-inflammatory, antimicrobial and cytotoxic properties of fungal endophytes from Garcinia species. Int J Pharm Pharm Sci 2013;5(3):889-97.
33. Devi NN, Prabakaran JJ, Wahab F. Phytochemical analysis and enzyme analysis of endophytic fungi from Centella asiatica. Asian Pac J Trop Biomed 2012;2(3):1280-84.
34. Sharma R, Kumar BS. Isolation characterization and antioxidant potential of endophytic fungi of Ocimum sanctum Linn. Indian J Appl Res 2013;3(7):5-10.
35. Kekuda TP, Prashith TR, Shobha KS, Onkarappa R. Potent insecticidal activity of two Streptomyces species isolated from the soils of Western ghats of Agumbe, Karnataka. J Nat Pharm 2010;1(1):30-2.
36. Zheng LP, Gao LW, Zhou JQ, Sima YH, Wang JW. Antioxidant activity of aqueous extract of a Tolypocladium sp fungus isolated from wild Cordyceps sinensis. Afr J Biotechnol 2008;7(17):3004-10.
37. Govindappa M, Bharath N, Shruthi HB, Santoyo G. In vitro antioxidant activity and phytochemical screening of endophytic extracts of Crotalaria pallida. Free Radic Antioxid 2011;1(3):79-86.
38. Lopes GK, Schulman HM, Hermes-Lima M. Polyphenol tannic acid inhibits hydroxyl radical formation from Fenton reaction by complexing ferrous ions. BBA Gen Subj 1999;1472(1-2):142-52.
39. Song TY, Yen GC. Antioxidant properties of Antrodia camphorata in submerged culture. J Agric Food Chem 2002;50(11):3322-7.
40. Qiu M, Xie RS, Shi Y, Zhang H, Chen HM. Isolation and identification of two flavonoid-producing endophytic fungi from Ginkgo biloba L. Ann Microbiol 2010;60(1):143-50.
41. Hata K, Futai K, Tsuda M. Seasonal and needle age-dependent changes of the endophytic mycobiota in Pinus thunbergii and Pinus densiflora needles. Can J Bot 1998;76(2):245-50.
42. Bors W, Saran M, Elstner EF. Screening for plant antioxidants. In: Plant Toxin Analysis. Berlin, Heidelberg: Springer; 1992. p. 277-95.
43. Wolfe K, Wu X, Liu RH. Antioxidant activity of apple peels. J Agric Food Chem 2003;51(3):609-14.
44. Basile A, Giordano S, LÃ³pez-SÃ¡ez JA, Cobianchi RC. Antibacterial activity of pure flavonoids isolated from mosses. Phytochemistry 1999;52(8):1479-82.
The publication is licensed under CC By and is open access. Copyright is with author and allowed to retain publishing rights without restrictions.