ANTIFUNGAL ACTIVITY OF ENDOPHYTIC FUNGI ISOLATED FROM LANNEA COROMANDELICA– AN INSILICO APPROACH Original Article N. PREMJANU a* , C. JAYNTHY b , S. DIVIYA b a Department of Biochemistry, Dental College and Hospital, Sathyabama University, Chennai, b Department of Bioinformatics, Sathyabama University, Chennai Email: janu6kishore@gmail.com Received: 24 Oct 2015 Revised and Accepted: 30 Mar 2016 ABSTRACT Objective: The objective of this research was to isolate endophytic fungi from Lannea coromandelica having antifungal activity potential and to isolate the secondary metabolite from the dominant fungi and predict the probable mechanism behind its activity. Methods: The endophytic fungi were isolated from leaves of Lannea coromandelica by surface sterilization method. Then fungal biomass was extracted for intracellular metabolites by using ethyl acetate as solvent. The crude extract was filtered, and the filtrate was dried under vacuum at 40 °C. The filtrate was analysed for antifungal activity. The fungi which showed the maximum activity was identified and the metabolite present in the ethyl acetate extract was characterized and identified by GC-MS (Gas-Chromatography Mass-spectrophotometry) analysis. Further, these compounds were docked against the target protein Lanosterol 14-alpha demethylase to unravel and predict the probable mechanism behind the antifungal activity of secondary metabolite. Results: Aspergillus flavus, Aspergillus niger, Alternaria alternata and Colletotrichum gloeosporioides were isolated and identified based on their morphological features as endophytic fungi. Among the four dominant fungi, the antifungal activity of Aspergillus flavus showed the maximum activity with an inhibitory zone of 26.22 mm against Candida albicans and 16.72 mm against Malassezia pachydermis. Further, the secondary metabolite was identified by GC-MS (Gas-Chromatography Mass-spectrophotometry) () analysis and found to be Kojic Acid, Octadecanoic acid, n- Hexadecanoic Acid, diethyl Phthlate, 3-Phenyl Propionic Acid. These compounds were docked with the target protein and were able to bind at an active site similar to that of Flucanozole a known inhibitor. Conclusion: The finding of this work clearly indicates that the metabolite produced by the endophytic fungus could be used as an alternative source of antifungal agents against clinical pathogens. Keywords: Endophytic fungi, Secondary metabolite, Antifungal activity © 2016 The Authors. Published by Innovare Academic Sciences Pvt Ltd. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/) INTRODUCTION In the last 15 y, there has been an alarming increase in life- threatening systemic fungal infections of varying types which are observed in immunosuppressed/immunocompromised person such as AIDS, cancer, and transplant patients. Candida sp. have emerged as significant pathogens in recent decades as a consequence of increasing numbers and population of immunocompromised patients [1]. As a result, they have become a major direct cause of death in patients being treated for malignant diseases and emerging immune deficiency diseases [2]. Management of fungal infections have become more complex due to lack of a number of effective antifungal drugs, problems of drug safety, side effects, resistance and effectiveness of drug forms [3]. Therefore, there is an urgent need to improve currently used drugs, to design new drugs and to explore an alternate source of novel drugs through various mechanisms, for better treatment Endophytic fungi are a group of microorganism colonized in inter and intracellular host plants without causing any disease, plays important physiological roles and ecological roles [4]. After colonization many fungal endophytes may occupy a large area relatively and create an obstacle for other endophytic species by competing [5] or by producing antagonistic metabolites [6, 7]. After the discoveries of endophytes which reside in unique biological niche that is intracellular cells of the plants, created a worldwide curiosity among scientist which led to isolation of endophytes and to study their natural products. It has been observed that plants harbour a myriad of endophytes, possessing enormous metabolic diversity and diverse bioactive compounds that can be used as a therapeutic agent against emerging human diseases such as cancer and infectious diseases [8, 9]. It has been reported that mainly the filamentous fungi, are capable of producing a large number of chemically different secondary metabolites [10], which possess diversified functions and can be used as antimicrobials, antifungal and antivirals. These metabolites have high chemical diversity which comprises of alkaloids, terpenoids, quinones, peptides, xanthones and phenols [4]. Due to the presence of the above-mentioned properties, it may provide numerous lead compounds for the development of new drugs, directly, or as an inspiration for synthetic drugs [8]. A considerable body of research has investigated the diversity, ecological role, secondary metabolites and bioactivity of the endophytic fungi isolated from various medicinal plants [11]. One such plant studied is Lannea coromandelica, which is commonly known as “The Indian Ash Tree” belonging to the Anacardiaceae family. The bark and leaves of this tree have been used to treat various aliment by villagers and tribal people. Lannea coromandelica is considered to have antimicrobial, antifungal and antiviral properties [12]. Apart from this, it is claimed to have anti- inflammatory, zoosporicidal, tissue healing, and tissue regeneration capacity. It is also used in the treatment of skin diseases, injuries, diarrhoea, particularly in young children, ulcers, impotence, vaginal troubles, heart diseases, gout, rheumatism, sore eyes, dysentery, and as an antidote in coma caused by the narcotics [13-15]. In this study an attempt was made to investigate and identify the antifungal metabolite produced by the fungal endophytes by GC-MS (Gas-Chromatography Mass-Spectrophotometry) analysis. Further studies were carried out to evaluate the interactions of metabolites present in the ethyl acetate extract of Aspergillus flavus by using In silico docking method for antifungal analysis with Lanosterol 14- alpha demethylase as target protein. Our study dealt with the assessment of the interactions between ligands and target protein by molecular docking method in order to calculate the minimum binding energy (kcal/mol) between them. International Journal of Pharmacy and Pharmaceutical Sciences ISSN- 0975-1491 Vol 8, Issue 5, 2016