Bioactivity Screening and Identification of Secondary Metabolites from Fungal Endophytes of Carica papaya L. Leaves


Affiliations

  • BCSIR Laboratories, Pharmaceutical Sciences Research Division, Dhaka, 1205, Bangladesh
  • Dr. Mahbubur Rahman Mollah College, Dhaka, 1362, Bangladesh
  • BCSIR Laboratories, Biological Research Division, Dhaka, 1205, Bangladesh
  • University of Dhaka, Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Dhaka, 1000, Bangladesh

Abstract

Objectives: This work describes the chemical and in-vitro bioactivity studies on endophytic fungi derived from the leaves of Carica papaya L. Methods: Column Chromatography was employed for isolation of fungal metabolites. Nuclear Magnetic Resonance (NMR) spectral data was utilized to elucidate the structure of the isolated compounds and physicochemical properties of them were also examined. As a part of in-vitro bioactivity screening disc diffusion method, 1, 1-diphenyl-2- picryl-hydrazyl (DPPH) scavenging assay and brine shrimp lethality bioassay were conducted to evaluate antimicrobial, antioxidant and cytotoxic activities, respectively. Results: A total of four fungal isolates were identified as endophytic fungi and purified from the young leaves of Carica papaya. These fungi, encoded as CPLE-1, CPLE-2, CPLE-3 and CPLE-4, were identified up to the genus level on the basis of their macroscopic and microscopic characteristics as Colletotrichum sp., Carvularia sp., and next two as Alternaria sp., respectively. Chemical profiling and preliminary bioactivity screening of the fungal endophytes signified strain CPLE-3 with highest potentiality to produce bioactive compounds which directs its large scale cultivation. Ergosterol and Alternariol monomethyl ether were isolated from the ethyl acetate extract of the fungus, CPLE-3. Conclusion: Bioactivity and chemical screening of the endophytic fungal extracts of Carica papaya leaf along with the isolation of fungal secondary metabolites suggested these endophytic fungi to be possible source of bioactive leads for developing new and improved medicines.

Keywords

Colletotrichum, Carvularia, Alternaria, Ergosterol, Alternariol Monomethyl Ether

Subject Discipline

Endophytic fungi; Medicinal plant investigation

Full Text:

References

Strobel G, Daisy B. Bioprospecting for microbial endophytes and their natural products. Microbiology and

Molecular Biology Reviews. 2003; 67(4):491–502. https://doi.org/10.1128/MMBR.67.4.491-502.2003. PMid:14665674. PMCid:PMC309047

Sathasivam K, Ramanathan S, Mansor SM, Haris MR, Wernsdorfer WH. Thrombocyte count in mice after administration of papaya leaf suspension. Wiener klinische Wochenschrift. 2009; 121:19–22. https://doi.org/10.1007/ s00508-009-1229-0. PMid:19915811

Romasi EF, Karina J, Parhusip AJN. Antibacterial activity of Papaya leaf extract against pathogenic bacteria. Makara Journal of Technology. 2011; 15:173–77. https://doi.org/10.7454/mst.v15i2.936

Chávez-Quintal P, González-Flores T, Rodríguez-Buenfil I, Santiago Gallegos-Tintoré S. Antifungal activity in ethanolic extracts of Carica papaya L. cv. Maradol leaves and seeds. Indian Journal of Applied Microbiology. 2011; 51:54–60. https://doi.org/10.1007/s12088-011-0086-5. PMid:22282629. PMCid:PMC3209867

Kusemiju TO, Yama OE, Okanlawon AO. Effect of Carica papaya bark extracts on oxidative stress parameters in testes of male albino mice. International Journal of Applied Research. 2012; 4:1–6.

Yudoh FV, Yudoh PB, Yumoh PF. Activity of alkaloid extract of Carica papaya seeds on reproductive functions in male Wister rats. Pharmaceutical Biology. 2005; 43:563–7. https://doi.org/10.1080/13880200500220961

Eze PM, Abonyi DO, Abba CC, Proksch P, Okoye FC, Esimone CO. Toxic, but beneficial compounds from endophytic fungi of Carica papaya EuroBiotech Journal. 2019; 3(2):105–11. https://doi.org/10.2478/ebtj-2019-0012

Abonyi DO, Eze PM, Abba CC, Chukwunwejim CR, Ejikeugwu CP, Okoye FB, Esimone CO. Metabolites of endophytic Colletotrichum gloeosporioides isolated from leaves of Carica papaya American Journal of Essential Oils and Natural Products. 2019; 7(1):39–46.

Rashmi M, Meena H, Meena C, Kushveer JS, Busi S, Murali A, Sarma VV. Anti-quorum sensing and antibiofilm potential of Alternaria alternata, a foliar endophyte of Carica papaya, evidenced by QS assays and in-silico analysis. Fungal Biology. 2018; 122(10):998–1012. https://doi.org/10.1016/j.funbio.2018.07.003. PMid:30227935

Wang H, Eze PM, Höfert SP, Janiak C, Hartmann R, Okoye FB, Esimone CO, Orfali RS, Dai H, Liu Z, Proksch P. Substituted L-tryptophan-L-phenyllactic acid conjugates produced by an endophytic fungus Aspergillus aculeatus using an OSMAC approach. RSC Advances. 2018; 8(14):7863–72. https://doi.org/10.1039/C8RA00200B

Kusari S, Lamshöft M, Spiteller M. Aspergillus fumigatus Fresenius, an endophytic fungus from Juniperus communis L. Horstmann as a novel source of the anticancer proâ€drug deoxypodophyllotoxin. Journal of Applied Microbiology. 2009; 107(3):1019–30. https://doi.org/10.1111/j.1365-2672.2009.04285.x. PMid:19486398

Sadananda S, Govindappa M, Ramachandra YL. In Vitro antioxidant activity of Lectin from different endophytic fungi of Viscum album L. British Journal of Pharmaceutical Research. 2014; 4(5):626–43. https://doi.org/10.9734/ BJPR/2014/6702

Devi NN, Prabakaran JJ. Bioactive metabolites from an endophytic fungus Penicillium sp. isolated from Centella asiatica. Current Research in Environmental & Applied Mycology. 2014; 4(1):34–43. https://doi.org/10.5943/ cream/4/1/3

Barnett HL, Hunter BB. 1972. Illustrated genera of imperfect fungi. 4th ed. Minneapolis: Burgess Publishing Co; 1972.

Kidd S, Halliday CL, Alexiou H, Ellis DH. Descriptions of Medical Fungi. Adelaide: Newstyle Printing; 2016.

Chowdhury NS, Sohrab MH, Rony SR, Sharmin S, Begum MN, Rana MS, Hasan CM. Identification and bioactive potential of endophytic fungi from Monochoria hastata (L.) Solms. Bangladesh Journal of Botany. 2016; 45(1):187–93.

Li G, Kusari S, Kusari P, Kayser O, Spiteller M. Endophytic Diaporthe sp. LG23 produces a potent antibacterial tetracyclic triterpenoid. Journal of Natural Products. 2015; 78(8):2128– 32. https://doi.org/10.1021/acs.jnatprod.5b00170. PMid:26186257

Alzoreky NS, Nakahara K. Antibacterial activity of extracts from some edible plants commonly consumed in Asia. Int J Food Microbiol. 2003; 80(3):223-30. https://doi.org/10.1016/S0168-1605(02)00169-1

Bauer AW, Kirby WM, Sherris JC, Truck M. Antibiotic susceptibility testing by a standard single disc method. American Journal of Clinical Pathology. 1966; 45:493–6. https://doi.org/10.1093/ajcp/45.4_ts.493. PMid:5325707

Williams WB, Cuvelier ME, Berset C. Use of a free radical method to evaluate antioxidant activity. Lebensmittel Wissenschraft und Technologie. 1995; 28(1):25–30. https://doi.org/10.1016/S0023-6438(95)80008-5

Meyer BN, Ferrighi NR, Putnam JE, Jacobson LB, Nichols DE, Mclaughlin JL. Brine Shrimp, a convenient general biossay for active plant constituents. Planta Medica. 1982; 45:31–4. https://doi.org/10.1055/s-2007-971236

Zhao J, Zhou L, Wang J, Shan T, Zhong L, Liu X, Gao X. Endophytic fungi for producing bioactive compounds originally from their host plants. In: MeÌndez-Vilas A, Ed, Current research, technology and education topics in applied microbiology and microbial biotechnology. Badajoz: Formatex Research Center; 2010. p. 567–76.

Rai M, Agarkar G, Rathod D. Multiple applications of endophytic Colletotrichum species occurring in medicinal plants. In: Guribâ€Fakim A, Ed, Novel plant bioresources: applications in food, medicine and Cosmetics. Newjersy: John Wiley & Sons, Inc; 2014. p. 227–36. https://doi.org/10.1002/9781118460566.ch18

Nwachukwu CU, Ngwoke KG, Eze PM, Eboka CJ, Okoye FB. Secondary metabolites from Curvularia sp, an endophytic fungus isolated from the leaves of Picralima nitida Durand and Hook (Apocynaceae). Tropical Journal of Natural Product Research. 2018; 2(5):209–13. https://doi.org/10.26538/tjnpr/v2i5.2

Kaaniche F, Hamed A, Abdel-Razek AS, Wibberg D, Abdissa N, El Euch IZ, Allouche N, Mellouli L, Shaaban M, Sewald N. Bioactive secondary metabolites from new endophytic fungus Curvularia sp isolated from Rauwolfia macrophylla. PloS one. 2019; 14(6): e0217627. https://doi.org/10.1371/journal.pone.0217627. PMid:31247016. PMCid:PMC6597039

Khiralla A, Spina R, Saliba S, Laurain-Mattar D. Diversity of natural products of the genera Curvularia and Bipolaris. Fungal Biology Reviews. 2019; 33(2):101–22. https://doi.org/10.1016/j.fbr.2018.09.002

Aly AH, Edrada-Ebel R, Indriani ID, Wray V, Müller WE, Totzke F, Zirrgiebel U, Schächtele C, Kubbutat MH, Lin WH, Proksch P. Cytotoxic metabolites from the fungal endophyte Alternaria sp. and their subsequent detection in its host plant Polygonum senegalense. Journal of Natural Products. 2008; 71(6):972–80. https://doi.org/10.1021/np070447m. PMid:18494522

Fang ZF, Yu SS, Zhou WQ, Chen XG, Ma SG, Li Y, Qu J. A new isocoumarin from metabolites of the endophytic fungus Alternaria tenuissima (Nees & T. Nees: Fr.) Wiltshire. Chinese Chemical Letters. 2012; 23(3):317–20. https://doi.org/10.1016/j.cclet.2011.11.021

Hellwig V, Grothe T, Mayer-Bartschmid AN, Endermann R, Geschke FU, Henkel T, Stadler M. Altersetin, a new antibiotic from cultures of endophytic Alternaria spp. Journal of Antibiotics. 2002; 55(10):881–92. https://doi.org/10.7164/ antibiotics.55.881. PMid:12523821

Lou J, Yu R, Wang X, Mao Z, Fu L, Liu Y, Zhou L. Alternariol 9-methyl ether from the endophytic fungus Alternaria sp. Samif01 and its bioactivities. Brazilian Journal of Microbiology. 2016; 47:96–101. https://doi.org/10.1016/j.bjm.2015.11.004. PMid:26887231. PMCid:PMC4822747

Goulston G, Mercer EI, Goad LJ. The identification of 24-methylene-24, 25-di hydrolanosterol and other possible ergosterol precursors in Phycomyces blakesleeanus and Agaricus campestris. Phytochemistry. 1975; 14(2):457–62. https://doi.org/10.1016/0031-9422(75)85109-0

Khan N, Afroz F, Begum MN, Rony SR, Sharmin S, Moni F, Hasan CM, Shahaa K, Sohrab MH. Endophytic Fusarium solani: A rich source of cytotoxic and antimicrobial napthaquinone and aza-anthraquinone derivatives. Toxicology Reports. 2018; 5:970–6.https://doi.org/10.1016/j.toxrep.2018.08.016. PMid:30294556. PMCid:PMC6169151

Tan N, Tao Y, Pan J, Wang S, Xu F, She Z, Lin Y, Jones EBG. Isolation, structure elucidation and mutagenicity of four alternariol derivatives produced by mangrove endophytic fungus. Chemistry of Natural Compounds. 2008; 44:296–300. https://doi.org/10.1007/s10600-008-9046-7

Gu W. Bioactive metabolites from Alternaria brassicicola ML-P08, an endophytic fungus residing in Malus halliana. World Journal of Microbiology and Biotechnology. 2009; 25: 1677–83. https://doi.org/10.1007/s11274-009-0062-y

Coates LM, Johnson GI. Postharvest diseases of fruit and vegetables. In: Brown JF, Ogle HJ, editors. Plant pathogens and plant diseases. Armidale: Rockvale Publications; 1997. p. 533–48.

Lugtenberg BJJ, Caradus JR, Johnson LJ. Fungal endophytes for sustainable crop production. FEMS Microbiology Ecology. 2016; 92(12):1–17. https://doi.org/10.1093/femsec/ fiw194. PMid:27624083


Refbacks

  • There are currently no refbacks.