Bioactivity Screening of Endophytic Fungus Eutypa linearis isolated from Coleus amboinicus (Lour.)
Keywords:
Eutypa linearis, submerged fermentation, cytotoxicity, antioxidant, endophyte.
Abstract
Coleus amboinicus (Lour.) is a medicinal plant containing various bioactive compounds. Endophytes are microorganisms living inside intracellular tissue of plants and known as the source of bioactive compounds. In order to explore the potential of endophyte in producing novel bioactive compounds, this study focused on isolating endophytic fungi from the leaves of C.amboinicus, characterisation and screening their metabolite bioactivity during submerged culture fermentation. Isolation of endophytic fungi from the leaves segment of C.amboinicus was conducted on PDA media and fungus identification was carried out by analyzing its morphology and molecular examination. The production of metabolites was examined using thin layer chromatography and gas chromatography. Identification of the endophytic fungus showed 98.84% similarity with Eutypa linearis. This species was fermented submergedly in PDB medium for 14 days under dark and exposed to light and the fermentation broth was extracted using ethyl acetate. The results showed that exposure to the light did not significantly influence metabolite production. The ethyl acetate extract exhibited antioxidant and cytotoxic activities. Antioxidant activity of this extract as examined by DPPH assay showed IC50 of 105.31 ± 2.11 µg/mL. Cytotoxic activity against Hela cell line was known to be the best among other cell lines (IC50 301.53 ± 11.34 µg/mL) although it was found to be non selective (SI<1). The extract contained five major compounds namely Benzenemethanol, 4-nitro-(CAS) p-Nitrobenzyl alcohol; 2-Pentadecanone (CAS) Pentadecan-2-one; (1R*,6S*,10R*)-5,5-Dimethyl-11,12-dioxatricyclo[8.2.1.0(1,6)] tridecan-10-ol; 9,12-Octadecadienoic acid (Z,Z)-, methyl ester (CAS) Methyl linoleate; and 3-Furanacetic acid, 4-hexyl-2,5-dihydro-2,5-dioxo- (CAS) 2-carboxymethyl-3-N-hexyl-maleic anhydride.
References
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Mosmann, T. (1983). Rapid Colorimetric Assay for Cellular Growth and Survival: Application to Proliferation and. Journal of Immunological Methods, 65, 55–63.
Patel, R., Mahobia, N., Waseem, N., Upwar, N., & Singh, S. (2010). Phyto-Physicochemical Investigation of Leaves of Plectranthus amboinicus (Lour) Spreng. Pharmacognosy Journal, 2(13), 536–542. https://doi.org/10.1016/S0975-3575(10)80057-4
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Samson, R. A. (1995). Introduction to Food-borne Fungi. Centraalbureau voor Schimmelcultures.
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Swamy, M. K., Arumugam, G., Kaur, R., Ghasemzadeh, A., Yusoff, M. Mohd., & Sinniah, U. R. (2017). GC-MS Based Metabolite Profiling, Antioxidant and Antimicrobial Properties of Different Solvent Extracts of Malaysian Plectranthus amboinicus Leaves. Evidence-Based Complementary and Alternative Medicine : ECAM, 2017. https://doi.org/10.1155/2017/1517683
Webster, J. (1980). Introduction to Fungi (2nd edition). Cambridge University Press.
Wiji Prasetyaningrum, P., Bahtiar, A., & Hayun, H. (2018). Synthesis and Cytotoxicity Evaluation of Novel Asymmetrical Mono-Carbonyl Analogs of Curcumin (AMACs) against Vero, HeLa, and MCF7 Cell Lines. Scientia Pharmaceutica, 86(2). https://doi.org/10.3390/scipharm86020025
Alam, Md. Z., Bari, Md. N., Muyibi, S. A., Jamal, P., & Al Mamun, A. (2011). Development of Culture Inoculum for Scale-Up Production of Citric Acid from Oil Palm Empty Fruit Bunches by Aspergillus niger. Procedia Environmental Science, 8, 396–402.
Arumugam, G., Swamy, M. K., & Sinniah, U. R. (2016). Plectranthus amboinicus (Lour.) Spreng: Botanical, Phytochemical, Pharmacological and Nutritional Significance. Molecules (Basel, Switzerland), 21(4), 369. https://doi.org/10.3390/molecules21040369
Astuti, P., Sudarsono, S., Nisak, K., & Nugroho, G. W. (2014). Endophytic Fungi Isolated from Coleus amboinicus Lour Exhibited Antimicrobial Activity. Advanced Pharmaceutical Bulletin; EISSN 2251-7308. https://doi.org/10.5681/apb.2014.088
Barnett, H. L., & Hunter, B. B. (1998). Illustrated Genera of Imperfect Fungi (4th Edition). Prentice-Hall, Inc.
Bhatt, P., & Negi, P. S. (2012). Antioxidant and Antibacterial Activities in the Leaf Extracts of Indian Borage (Plectranthus amboinicus). Food and Nutrition Sciences, 3(2), 146–152. https://doi.org/10.4236/fns.2012.32022
Chen, L., Zhang, Q.-Y., Jia, M., Ming, Q.-L., Yue, W., Rahman, K., Qin, L.-P., & Han, T. (2016). Endophytic fungi with antitumor activities: Their occurrence and anticancer compounds. Critical Reviews in Microbiology, 42(3), 454–473. https://doi.org/10.3109/1040841X.2014.959892
de Torre, M. P., Cavero, R. Y., Calvo, M. I., & Vizmanos, J. L. (2019). A Simple and a Reliable Method to Quantify Antioxidant Activity In Vivo. Antioxidants, 8(5), 142. https://doi.org/10.3390/antiox8050142
Domsch, K. H., Gams, W., & Anderson, T.-H. (1980). Compendium of Soil Fungi. Academic Press.
Ellis, M. B. (1971). Dematiaceous Hyphomycetes. Commonwealth Mycological Institute.
Geran, R. I., N.H, G., & M.M, M. D. (1972). Protocol for Screening Chemical Agents and Natural Products Against Animal Tumors and other Biologycal Systems. 3(2), 1–103.
Gouda, S., Das, G., Sen, S. K., Shin, H.-S., & Patra, J. K. (2016). Endophytes: A Treasure House of Bioactive Compounds of Medicinal Importance. Frontiers in Microbiology, 7. https://doi.org/10.3389/fmicb.2016.01538
Gunasekaran, S., & Poorniammal, R. (2008). Optimization of fermentation conditions for red pigment production from Penicillium sp. Under submerged cultivation. African Journal of Biotechnology, 7(12), 1894–1898. https://doi.org/10.5897/AJB2008.000-5037
Hardoim, P. R., Overbeek, L. S. van, Berg, G., Pirttilä, A. M., Compant, S., Campisano, A., Döring, M., & Sessitsch, A. (2015). The Hidden World within Plants: Ecological and Evolutionary Considerations for Defining Functioning of Microbial Endophytes. Microbiology and Molecular Biology Reviews, 79(3), 293–320. https://doi.org/10.1128/MMBR.00050-14
Hasibuan, P. A. Z., Ilyas, S., & Nasution, M. P. (2013). Antioxidant and Cytotoxic Activities of Plectranthus amboinicus (Lour.) Spreng. Extracts. International Journal of Pharmacy Teaching and Practice, 4(3), 755–758.
Hawksworth, D. L. (1974). Mycologist’s Handbook: An Introduction to the Principles of Taxonomy and Nomenclature in the Fungi and Lichens. Commonwealth Mycological Institute.
Jalgaonwala, R., Mohite, B. V., & Mahajan, R. (2011). A review: Natural products from plant associated endophytic fungi. J. Microbiol. Biotechnol. Res, 1, 21–32.
Jork, H. (Ed.). (1990). Thin-layer chromatography: Reagents and detection methods. VCH.
Kedare, S. B., & Singh, R. P. (2011). Genesis and development of DPPH method of antioxidant assay. Journal of Food Science and Technology, 48(4), 412–422. https://doi.org/10.1007/s13197-011-0251-1
Kumar, P., Singh, B., Thakur, V., Thakur, A., Thakur, N., Pandey, D., & Chand, D. (2019). Hyper-production of taxol from Aspergillus fumigatus, an endophytic fungus isolated from Taxus sp. Of the Northern Himalayan region. Biotechnology Reports, 24. https://doi.org/10.1016/j.btre.2019.e00395
Kumaran, A., & Joel karunakaran, R. (2006). Antioxidant and free radical scavenging activity of an aqueous extract of Coleus aromaticus. Food Chemistry, 97(1), 109–114. https://doi.org/10.1016/j.foodchem.2005.03.032
Lin, Z.-J., Lu, Z.-Y., Zhu, T.-J., Fang, Y.-C., Gu, Q.-Q., & Zhu, W.-M. (2008). Penicillenols from Penicillium sp. GQ-7, an endophytic fungus associated with Aegiceras corniculatum. Chemical & Pharmaceutical Bulletin, 56(2), 217–221. https://doi.org/10.1248/cpb.56.217
M. Blackwell Constantine J. Alexopoulos, C. W. M. (2007). Introductory Mycology, 4Th Ed (4th edition). Wiley.
Manjamalai, A. (2012). Volatile constituents and antioxidant property of essential oil from Plectranthus amboinicus (Lour). Int. J. Pharm. Biol. Sci, 3, 445–458.
Mosmann, T. (1983). Rapid Colorimetric Assay for Cellular Growth and Survival: Application to Proliferation and. Journal of Immunological Methods, 65, 55–63.
Patel, R., Mahobia, N., Waseem, N., Upwar, N., & Singh, S. (2010). Phyto-Physicochemical Investigation of Leaves of Plectranthus amboinicus (Lour) Spreng. Pharmacognosy Journal, 2(13), 536–542. https://doi.org/10.1016/S0975-3575(10)80057-4
Phongpaichit, S., Nikom, J., Rungjindamai, N., Sakayaroj, J., Hutadilok-Towatana, N., Rukachaisirikul, V., & Kirtikara, K. (2007). Biological activities of extracts from endophytic fungi isolated from Garcinia plants: Biological activities of extracts from endophytic fungi. FEMS Immunology & Medical Microbiology, 51(3), 517–525. https://doi.org/10.1111/j.1574-695X.2007.00331.x
Porras-Alfaro, A., & Bayman, P. (2011). Hidden Fungi, Emergent Properties: Endophytes and Microbiomes. Annual Review of Phytopathology, 49(1), 291–315. https://doi.org/10.1146/annurev-phyto-080508-081831
Prayong, P., Barusrux, S., & Weerapreeyakul, N. (2008). Cytotoxic activity screening of some indigenous Thai plants. Fitoterapia, 79(7–8), 598–601. https://doi.org/10.1016/j.fitote.2008.06.007
Samson, R. A. (1995). Introduction to Food-borne Fungi. Centraalbureau voor Schimmelcultures.
Singh, G., Singh, O. P., Prasad, Y. R., Lampasona, M. P. de, & Catalan, C. (2002). Studies on essential oils, Part 33: Chemical and insecticidal investigations on leaf oil of Coleus amboinicus Lour. Flavour and Fragrance Journal, 17(6), 440–442. https://doi.org/10.1002/ffj.1123
Swamy, M. K., Arumugam, G., Kaur, R., Ghasemzadeh, A., Yusoff, M. Mohd., & Sinniah, U. R. (2017). GC-MS Based Metabolite Profiling, Antioxidant and Antimicrobial Properties of Different Solvent Extracts of Malaysian Plectranthus amboinicus Leaves. Evidence-Based Complementary and Alternative Medicine : ECAM, 2017. https://doi.org/10.1155/2017/1517683
Webster, J. (1980). Introduction to Fungi (2nd edition). Cambridge University Press.
Wiji Prasetyaningrum, P., Bahtiar, A., & Hayun, H. (2018). Synthesis and Cytotoxicity Evaluation of Novel Asymmetrical Mono-Carbonyl Analogs of Curcumin (AMACs) against Vero, HeLa, and MCF7 Cell Lines. Scientia Pharmaceutica, 86(2). https://doi.org/10.3390/scipharm86020025
Published
2021-03-15
How to Cite
Gemantari, B. M., Romadhonsyah, F., Nurrochmad, A., Wahyuono, S., & Astuti, P. (2021). Bioactivity Screening of Endophytic Fungus Eutypa linearis isolated from Coleus amboinicus (Lour.). Indonesian Journal of Pharmacy, 32(1), 86-95. https://doi.org/10.22146/ijp.1077
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Research Article
