In Vitro Anti-HIV Activity of Partially Purified Coumarin(s) Isolated from Fungal Endophyte, Alternaria Species of Calophyllum inophyllum

Melappa Govindappa^1*, Kavya C. Hemmanur¹, S. Nithin¹, Chandrappa Chinna Poojari¹, Gopalakrishna Bhat², K. Channabasava¹

¹Endophytic Natural Product Laboratory, Department of Biotechnology, Shridevi Institute of Engineering & Technology, Tumkur, India.
²Madhuca, Srinivasa Nagara, Chitpady, Udupi, India.

DOI: 10.4236/pp.2015.67034                

Abstract

5 totally different endophytic fungal species were isolated from bark and leaf parts of Calophyllum inophyllum. Leaf part yielded Trichoderma harzianum and Alternaria species, whereas bark showed the presence of Fusarium species, Aspergillus species and unidentified fungi. Two solvents (hexane and methanol) were used for endophytic fungal extraction and the Alternaria species had shown the presence of coumarin whereas Trichoderma harzianum in methanol extract and Fusarium species in hexane extract had shown the coumarin(s) in all the four methods tested. The total coumarin yield was more in microwave assistance method, the methanol Alternaria species (3.941 ± 0.082) stood first, followed by hexane extract of Alternaria species (3.254 ± 0.082), Fusarium species (2.532 ± 0.082) and Trichoderma harzianum (2.294 ± 0.082), the plant extract showed 4.149 + 0.053. The methanol extract of Alternaria species inhibited the activity of HIV-Reverse Transcriptase (RT) (82.81 ± 1.0), integrase (98%) and protease (78) in maximum level followed by hexane extract of Alternaria species (71.12 ± 0.9, 89, 68), Fusarium species (63.92 ± 1.8, 67, 66) and Trichoderma harzianum (56.69 ± 0.9, 71, 63). The endophytic fungi Alternaria species inhibited all the three viral enzymes at maximum level and it was more than standard drug. However, in order to know possible anti-HIV, it is necessary to isolate active coumarin from the Alternaria species and the mechanism of action will be studied in future studies.

Keywords

Endophytes, Alternaria Species, HIV-RT, Integrase, Protease, Inhibitory Activity

Conflicts of Interest

The authors declare no conflicts of interest.

References

[1]	Jassim, S.A.A. and Naji, M.A. (2003) Novel Antiviral Agents: A Medicinal Plant Perspective. Journal of Applied Microbiology, 95, 412-427. http://dx.doi.org/10.1046/j.1365-2672.2003.02026.x
[2]	Narayan, C.L. and Rai, R.V. (2011) A Screening Strategy for Selection of Anti-HIV Integrase and anti-HIV Protease Inhibiotors from Plant Extracts of Indian Medicinal Plants. International Journal of Phytomedicine, 3, 312-318.
[3]	Tewtrakul, S., Nakamura, N., Hattori, M., Fujiwara, T. and Supavita, T. (2002) Flavanone and Flavonol Glycosides from the Leaves of Thevetia peruviana and Their HIV-1 Reverse Transcriptase and HIV-1 Integrase Inhibitory Activities. Chemical and Pharmaceutical Bulletin, 50, 630-635.
[4]	Sundur, S., Shrivastava, B., Sharma, P., Raj, S.S. and Jayasekhar, V.L. (2014) A Review Article of Pharmacological Activities and Biological Importance of Calophyllum inophyllum. International Journal of Advanced Research, 2, 599-603.
[5]	Umashankar, T., Govindappa, M. and Ramachandra, Y.L. (2014) In Vitro Antioxidant and Antimicrobial Activity of Partially Purified Coumarins from Fungal Endophytes of Crotalaria pallid. International Journal of Current Microbiology and Applied Sciences, 3, 58-72.
[6]	Jagessar, R.C. and Cox, M. (2010) Phytochemical Screening of the CHCl3 and CH3CH2OH Extract of Stems, Twigs, Roots and Barks of Conocarpus erectus L. International Journal of Academic Research, 2, 36-45.
[7]	Rege, A.A., Ambaye, R.Y. and Deshmukh, R.A. (2010) In Vitro Testing of Anti-HIV Activity of Some Medicinal Plants. Indian Journal of Natural Products and Resources, 1, 193-199.
[8]	Tewtrakul, S., Subhadhirasakul, S. and Kummee, S. (2003) HIV-1 Protease Inhibitory Effects of Medicinal Plants Used as Self Medication by AIDS Patients. Songklanakarin Journal of Science and Technology, 25, 239-243.
[9]	Tewtrakul, S., Miyashiro, H., Hattori, M., Yoshinaga, T., Fujiwara, T., Tomimori, T., et al. (2001) Inhibitory Effects of Flavonoids on Human Immunodeficiency Virus Type-1 Integrase. Journal of Traditional Medicines, 18, 229-238.
[10]	Govindappa, M., Kumar, N.V.A. and Santoyo, G. (2011) Crotalaria pallida Extracts as a Putative HIV-Protease Inhibitors. Journal of Research in Biology, 4, 285-291.
[11]	Estari, M., Venkanna, L., Sripriya, D. and Lalitha, R. (2012) Human Immunodeficiency Virus (HIV-1) Reverse Transcriptase Inhibitory Activity of Phyllanthus emblica Plant Extract. Biology and Medicine, 4, 178-182.
[12]	Rege, A.A. and Chowdhury, A.S. (2014) Evaluation of Ocimum sanctum and Tinospora cordifolia as Probable HIV Protease Inhibitors. International Journal of Pharmaceutical Sciences Review and Research, 25, 315-318.
[13]	Xiang, Z.C., Jiang, Y. and Guo, S.X. (2006) In Vitro Anti-HIV Activity of a Chinese Fungus Extract. Biomedical and Environmental Sciences, 19, 169-172.
[14]	Umashankar, T., Govindappa, M. and Ramachandra, Y.L. (2012) In Vitro Antioxidant and Anti-HIV Activity of Endophytic Coumarin from Crotalaria pallida Aiton. Planta Medica, 78, 102. http://dx.doi.org/10.1055/s-0032-1307610
[15]	Wellensiek, B.P., Ramakrishnan, R., Bashyal, B.P., Eason, Y., Gunatilaka, A.A.L. and Ahmad, N. (2013) Inhibition of HIV-1 Replication by Secondary Metabolites from Endophytic Fungi of Desert Plants. The Open Virology Journal, 7, 72-80. http://dx.doi.org/10.2174/1874357920130624002