Optimization of process parameters for production of antimicrobial metabolites by an endophytic fungus Aspergillus sp. CPR5 isolated from Calotropis procera root.

  • SHIV KR. VERMA School of Biochemical Engineering IIT - BHU Varanasi
  • Moti Lal


Objective:To study the antimicrobial activity of crud ethylacetate  extract from endophytic fungus in Calotropis procera root.

Methods:Endophytic fungus was screened for production of antimicrobial metablites.fermentation was carried out in 500 ml Erlenmeyer flask. Disc diffusion method was used to test antimicrobial activity of crude extract using chloramphenicol sulphate and Flucnazole as positive controle

Ressults: A total of fourteen Endophytic fungi were isolated (CPR1- CPR14). Among these fourteen isolates, CPR5 was found to show maximum antimicrobial activity, in compare to other isolates, against gram positive, gram negative bacteria and fungi. The antimicrobial activity was tested against Escherichia coli. Pseudomonas aeroginosa, Bacillus subtilis, Staphylococcus aureus, Ralstonia solanacearum, Xanthomonas oryzae, Penicillium chrysogenum, Candida albicans, Phoma exigua, Sclerotium rolfsii and Sclerotinia scleratiourum. Minimum inhibitory concentration (MIC) of crude extract against test microorganisms was determined. Fungus was identified as aspergillus sp. Prduction parameters (Temprature, pH, Carbon source, Nitrgen surce, Sdium Chlride concentration) were optimized.

Conclusion:  Crud  extract produced by the isolated endophytic fungus could be an important source of broad spectrum antimicrobial metabolites. 

Keywords: Endophytic fungi, Antimicrobial metabolites, Process optimization, Calotropis procera, Inhibition zone.

Author Biography

SHIV KR. VERMA, School of Biochemical Engineering IIT - BHU Varanasi

Research Scholar IIT BHU Varanasi


1. Petrini O, Stone J, Carroll FE. Endophytic fungi in evergreen shrubs in Western Oregon: A prelimenary study. Can J Bot 1982;60(6):789-96.
2. Gond SK, Mishra A, Sharma VK, Verma SK, Kharwar RN, Kumar A. Diversity and antimicrobial activity of endophytic fungi isolated from nyctanthes arbor-tristis, a well-known medicinal plant of India. Mycoscience 2012;53(2):113-21.
3. Begum MM, Sariah M, Puteh AB, Abidin MA. Pathogenicity of Colletotrichum truncatum and ITS influence on soybean seed quality. Int J Agric Biol 2008;10(4):393-8.
4. Amirita A, Sindhu P, Swetha J, Vasanthi NS, Kannan KP. Enumeration of endophytic fungi from medicinal plants and screening of extracellular enzymes. World J Microbiol Biotechnol 2012;2(2):13-9.
5. Strobel G, Daisy B. Bioprospecting for microbial endophytes and their natural products. Microbiol Mol Biol Rev 2003;67(4):491-502.
6. Verma SK, Kumar A, Verma A, Debnath MD. Antimicrobial activity of endophytic fungal isolate in Argemone maxicana; A traditional Indian medicinal plant. Int J Innov Res Sci Eng Technol 2014;3(3):10151-62.
7. Hawksworth DL, Rossman AY. Where are all the undescribed fungi? Phytopathology 1997;87(9):888-91.
8. Preetha S, Srinivasan V. Antioxidant activities of ropinirole and pramipexole novel drugs used in treatment of parkinsonism: An in vitro approach. Asian J Pharm Clin Res 2016;9(3):105-7.
9. Stierle A, Strobel GA, Stierle D. Taxol and taxen production by Taxomyces andreanae an endophytic Fungus of Pacific yew. Science 1993;260(5105):214-6.
10. Strobel G, Yang X, Sears J, Kramer R, Sidhu RS, Hess WM. Taxol from Pestalotiopsis microspora, an endophytic fungus of Taxus wallachiana. Microbiology 1996;142:435-40.
11. Kang JG, Shin SY, Kim MJ, Bajpai V, Maheshwari DK, Kang SC. Isolation and anti-fungal activities of 2-hydroxymethyl-chroman-4-one Produced by Burkholderia sp. MSSP. J Antibiot (Tokyo) 2004;57(11):726-31.
12. Myers N, Mittermeier RA, Mittermeier CG, da Fonseca GA, Kent J. Biodiversity hotspots for conservation priorities. Nature 2000;403(6772):853-8.
13. Gowrish A, Vagdevi HM, Rajashekar H. Phytochemical screening and antimicrobial activity of Leucas marrubioides desf. Root extracts. Int J Pharm Pharm Sci 2016;8(12):209-12.
14. Pharamat T, Palaga T, Piapukiew J, Anthony JS, Whalley AJS, Sihanonth P. Antimicrobial and anticancer activities of endophytic fungi from Mitrajyna javanica Koord and Val. Afr J Microbiol Res 2013;7(49):5565-72.
15. Sing AK, Raghubanshi AS, Sing JS. Medical ethnobotany of the tribals of Sonaghati of Sonbhadra district, Uttar Pradesh, India. J Ethnopharmacol 2002;81(1):31-41.
16. Parrotta JA. Healing Plants of Peninsular India. Wallingford, UK and New York: CAB International; 2001. p. 944.
17. Mossa JS. Pharmacological studies on aerial parts of Calotropis procera. Am J Chin Med 1991;19(3):223-31.
18. Anver S, Alam MM. Effect of latex seed dressing on interacting root-knot and reniform nematodes. Afro Asian J Nematol 1992;2(1):17-20.19. Petrini O, Sieber TN, Toti L, Viret O. Ecology, metabolite production, and substrate utilization in endophytic fungi. Nat Toxins 1992;1(3):185-96.
20. 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.
21. White TJ, Bruns TD, Lee SB, Taylor JW. Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. In: Innis MA, Gelfand DH, Sninshy JJ, White TD, editors. PCR Protocols: A Guide to Methods and Applications. London: Academic Press; 1990. p. 315-32.
22. Raahave D. Agar contact plate in evaluation of skin disinfection. Dan Med Bull 1974;20(6):204-8.
23. Saba Y, Vijay LM, Rajesh KM. Isolation and characterization of bioactive streptomyces from mangrove ecosystem of Machilipatnam, Krishna district, Andhra Pradesh. Asian J Pharm Clin Res 2016;9(3):258-63.
24. Arunodaya HS, Krishna V, Shashikumar R, Girish KK. Antibacterial and antioxidant activities of stem bark essential oil constituents of Litsea glutinosa C. B. Rob. Int J Pharm Pharm Sci 2016;8(12):258-64.
25. Hutter R. Design of culture media capable of provoking wide gene expression. In: Bullock JD, Nisbet LJ, Winstanley DJ, editors. Bioactive Microbial Products, Search and Discovery. London: Academic Press; 1982. p. 37-50.
26. Radu S, Kqueen CY. Preliminary screening of endophytic fungi from medicinal plants in Malayasia for antimicrobial and antitumour activity. Malays J Med Sci 2002;9(2):23-33.
27. Lethimaki J, Moisander P, Sivonen K, Kononen K. Growth, nitrogen fixation and nodularin production by two Baltic Sea cyenobacteria. Appl Environ Microbiol 1997;63(5):1647-54.
28. Moita C, Feio SS, Nunes L, Joa M, Curto M, Roseiro JC. Optimisation of physical factors on the production of active metabolites by Bacillus subtilis 355 against wood surface contaminant fungi. Int Biodeterior Biodegradation 2005;55(4):261-9.
29. Digrak M, Eluk SZ. Determination of some fungal metabolite as influenced by temperature, time, pH and sugars by bioassay method. Turk J Biol 2001;25:197-203.
30. Hansen OH. Ecology, physiology, and biochemistry of blue-green algae. Annu Rev Microbiol 1968;22:47-57.
31. Sailer M, Helms GL, Henkel T, Niemczura WP, Stiles ME, Vederas JC. 15N-and 13C-labeled media from Anabaena sp. For universal isotopic labeling of bacteriocins: NMR resonance assignments of leucocin A from Leuconostoc gelidum and nisin A from Lactococcus lactis. Biochemistry 1993;32(1):310-8.
32. Vahidi H, Kobarfard F, Namjoyan F. Effect of cultivation conditions on growth and antifungal activity of Mycena leptocephala. Afr J Biotechnol 2004;3(11):606-9.
33. Gogoi DK, Boruah HP, Saikia R, Bora TC. Optimization of process parameters for improved production of bioactive metabolite by a novel endophytic fungus Fusarium sp. DF2 isolated from Taxus wallichiana. Microbiol Biotechnol 2008;24(1):79-87.
334 Views | 706 Downloads
How to Cite
VERMA, S. K., M. Lal, and M. D. (DAS). “Optimization of Process Parameters for Production of Antimicrobial Metabolites by an Endophytic Fungus Aspergillus Sp. CPR5 Isolated from Calotropis Procera Root.”. Asian Journal of Pharmaceutical and Clinical Research, Vol. 10, no. 4, Apr. 2017, pp. 225-30, doi:10.22159/ajpcr.2017.v10i4.16631.
Original Article(s)