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Characterization and biological activities of melanin pigment from root endophytic fungus, Phoma sp. RDSE17

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Abstract

Melanins are high molecular weight hydrophobic pigments which have gained popularity for their role in virulence against different pathogens. In the present study, we isolated and characterized the melanin pigment produced by a dark septate endophyte fungus Phoma sp. RDSE17, which was associated with the roots of an indigenous Oryza sativa cv. 'Chakhao amubi' in Manipur, Northeast India. The biological properties of purified melanin from the fungus were evaluated for their antioxidant, antimicrobial and anticancerous activities. The pigment was extracted from Phoma sp. by alkaline–acid hydrolysis method and confirmed as melanin through physico-chemical tests and spectral (UV, FTIR, and EPR) analysis. The analyses of the elemental composition indicated that the pigment possessed a low percentage of nitrogen (N) contents, and therefore, would not fall under DOPA class of melanin. Exposure of the fungus to melanin pathway inhibitors revealed a positive melanin inhibition by tricyclazole, but not by kojic acid. Thus, the melanin from Phoma sp. may be a member of the DHN family. Moreover, the purified melanin showed high DPPH (1, 1-Diphenyl-2-picrylhydrazyl) free radical-scavenging activity with an EC50 of 69 µg/mL and inhibited human lung cancer cell (A549 cells) proliferation at 80 µg/mL. The present study demonstrates that melanin from Phoma sp. RDSE17 could be employed as a potential biological (antioxidant) and antimicrobial agent for inhibiting the growth of humans and phytopathogens.

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References

  • Addy HD, Piercey MM, Currah RS (2005) Microfungal endophytes in roots. Can J Bot 83:1–13

    Article  Google Scholar 

  • Akilandeswari P, Pradeep BV (2016) Exploration of industrially important pigments from soil fungi. Appl Microbiol Biotechnol 100(4):1631–1643

    Article  CAS  PubMed  Google Scholar 

  • Altschul SF, Madden TL, Schaffer AA, Zhang J, Zhang Z, Miller W, Lipman DJ (1997) Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucl Acids Res 25:3389–3402

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Arun G, Eyini M, Gunasekaran P (2015) Characterization and biological activities of extracellular melanin produced by Schizophyllum commune (Fries). Ind J Exp Biol 53:380–387

    CAS  Google Scholar 

  • Asem ID, Imotomba RK, Mazumder PB, Laishram JM (2015) Anthocyanin content in the black scented rice (Chakhao): its impact on human health and plant defense. Symbiosis 66(1):47–54

    Article  CAS  Google Scholar 

  • Aveskamp MM, de Gruyter J, Woudenberg JHC, Verkley GJM, Crous PW (2010) Highlights of the Didymellaceae: a polyphasic approach to characterise Phoma and related pleosporalean genera. Stud Mycol 65(1):1–60

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Badillo-Vargas IE, Rivera-Vargas LI, Calle-Bellido J (2008) Morphological, pathogenic and molecular characterization of Phoma spp. isolated from onion field soils in Puerto Rico. J Agric Univ PR 92(1–2):73–86.

  • Berthelot C, Perrin Y, Leyval C, Blaudez D (2017) Melanization and ageing are not drawbacks for successful agro-transformation of dark septate endophytes. Fun Biol 121:652–663

    Article  CAS  Google Scholar 

  • Bhatia D, Mittal A, Malik DK (2016) Antimicrobial activity of PVP coated silver nanoparticles synthesized by Lysinibacillus varians. 3 Biotech 6:196. https://doi.org/10.1007/s13205-016-0514-7

  • Boerema GH, de Gruyter J, Noordeloos ME, Hamers MEC (2004) Phoma identification manual: differentiation of specific and infra-specific taxa in culture. CABI Publishing, Wallingford, Oxford shire, UK

    Book  Google Scholar 

  • Dadachova E, Bryan RA, Huang X, Moadel T, Schweitzer AD, Aisen P, Nosanchuk JD, Casadevall A (2007) Ionizing radiation changes the electronic properties of melanin and enhances the growth of melanized fungi. PLoS ONE 2(5):e457. https://doi.org/10.1371/journal.pone.0000457

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • De Souza RA, Kamat NM, Nadkarni VS (2018) Purification and characterization of a sulphur rich melanin from edible mushroom Termitomyces albuminosus Heim. Mycology 9(4):296–306

    Article  PubMed  PubMed Central  Google Scholar 

  • Dong C, Yao Y (2012) Isolation, characterization of melanin derived from Ophiocordyceps sinensis, an entomogenous fungus endemic to the Tibetan Plateau. J Biosci Bioeng 113(4):474–479

    Article  CAS  PubMed  Google Scholar 

  • Dufossé L, Fouillaud M, Caro Y, Mapari SAS, Sutthiwong N (2014) Filamentous fungi are large scale producers of pigments and colorants for the food industry. Curr Opin Biotechnol 26:56–61

    Article  PubMed  Google Scholar 

  • Eisenman HC, Casadevall A (2012) Synthesis and assembly of fungal melanin. Appl Microbiol Biotechnol 93:931–940

    Article  CAS  PubMed  Google Scholar 

  • Enochs WS, Nilges MJ, Swartz HM (1993) A standardized test for the identification and characterization of melanins using electron-paramagnetic-resonance (EPR) spectroscopy. Pigment Cell Res 6:91–99

    Article  CAS  PubMed  Google Scholar 

  • Gadd GM (1982) Effects of media composition and light on colony differentiation and melanin synthesis in Microdochium bolleyi. Trans Br Mycol Soc 78:115–122

    Article  CAS  Google Scholar 

  • Giménez C, Cabrera R, Reina M, González-Coloma A (2007) Fungal endophytes and their role in plant protection. Curr Org Chem 11:707–720

    Article  Google Scholar 

  • Goncalves RC, Lisboa HC, Pombeiro-Sponchiado SR (2012) Characterization of melanin pigment produced by Aspergillus nidulans. World J Microbiol Biotechnol 28:1467–1474

    Article  CAS  PubMed  Google Scholar 

  • Hall T (1999) BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows95/98/NT. Nucl Acids Symp Ser 41:95–98

    CAS  Google Scholar 

  • Helan Soundra Rani M, Ramesh T, Subramanian J, Kalaiselvam M (2013) Production and characterization of melanin pigment from halophilic black yeast Hortaea werneckii. Int J Pharma Res Rev 2(8):9–17

    Google Scholar 

  • Hou R, Liu X, Xiang K, Chen L, Wu X, Lin W, Zheng M, Fu J (2019) Characterization of the physicochemical properties and extraction optimization of natural melanin from Inonotus hispidus mushroom. Food Chem 277:533–542. https://doi.org/10.1016/j.foodchem.2018.11.002

    Article  CAS  PubMed  Google Scholar 

  • Huang L, Liu M, Huang H, Wen Y, Zhang X, Wei Y (2018) Recent advances and progress on melanin-like materials and their biomedical applications. Biomacromol 19:1858–1868. https://doi.org/10.1021/acs.biomac.8b00437

    Article  CAS  Google Scholar 

  • Jacobson ES, Tinnell SB (1993) Antioxidant function of fungal melanin. J Bacteriol 175(21):7102–7104

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Kejžar A, Gobec S, Plemenitaš A, Lenassi M (2013) Melanin is crucial for growth of the black yeast Hortaea werneckii in its natural hypersaline environment. Funal Biol 117:368–379

    Article  Google Scholar 

  • Kim JW, Choi HG, Song JH, Kang KS, Shim SH (2018) Bioactive secondary metabolites from an endophytic fungus Phoma sp. PF2 derived from Artemisia princeps Pamp. J Antibiot 72(3):174–177.

  • Kumar CG, Mongolla P, Pombala S, Kamle A, Joseph J (2011) Physicochemical characterization and antioxidant activity of melanin from a novel strain of Aspergillus bridgeri ICTF-201. Lett Appl Microbiol 53:350–358

    Article  CAS  PubMed  Google Scholar 

  • Kusari S, Christian H, Michael S (2012) Chemical ecology of endophytic fungi: Origins of secondary metabolites. Chem Biol 19(7):792–798

    Article  CAS  PubMed  Google Scholar 

  • Li C, Ji C, Tang B (2018) Purification, characterisation and biological activity of melanin from Streptomyces sp. FEMS Microbiol Lett 365(19):1–8. https://doi.org/10.1093/femsle/fny077

    Article  CAS  Google Scholar 

  • Łopusiewicz L (2018) Isolation, characterisation and biological activity of melanin from Exidia nigricans. World Sci News 91:111–129

    Google Scholar 

  • Mandyam K, Jumpponen A (2005) Seeking the elusive function of the root-colonising dark septate endophytic fungi. Stud Mycol 53(1):173–189

    Article  Google Scholar 

  • Montefiori DC, Zhou JY (1991) Selective antiviral activity of synthetic and soluble L-tyrosine and L-DOPA melanin against human immunodeficiency virus in vitro. Antiviral Res 15:11–25

    Article  CAS  PubMed  Google Scholar 

  • Mosmann T (1983) Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods 65:55–63

    Article  CAS  PubMed  Google Scholar 

  • Naik BS, Shashikala J, Krishnamurthy YL (2009) Study on the diversity of endophytic communities from rice (Oryza sativa L.) and their antagonistic activities in vitro. Microbiol Res 164:290–296

    Article  CAS  PubMed  Google Scholar 

  • Newsham KK, Upson R, Read DJ (2009) Mycorrhizas and dark septate root endophytes in Polar Regions. Fungal Ecol 2(1):10–20

    Article  Google Scholar 

  • Nosanchuk JD, Casadevall A (2006) Impact of melanin on microbial virulence and clinical resistance to antimicrobial compounds. Antimicrob Agents Chemother 50:3519–3528

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Nosanchuk JD, Stark RE, Casadevall A (2015) Fungal melanin: what do we know about structure? Front Microbiol 6:1463. https://doi.org/10.3389/fmicb.2015.01463

    Article  PubMed  PubMed Central  Google Scholar 

  • Pal AK, Gajjar DU, Vasavada AR (2014) DOPA and DHN pathway orchestrate melanin synthesis in Aspergillus species. Med Mycol 52:10–18

    CAS  PubMed  Google Scholar 

  • Potshangbam M, Devi SI, Sahoo D, Strobel GA (2017) Functional characterization of endophytic fungal community associated with Oryza sativa L. and Zea mays L. Front Microbiol 8:325. https://doi.org/10.3389/fmicb.2017.00325

  • Rai MK, Deshmukh P, Gade A, Ingle A, Kövics GJ, Irinyi L (2009) Phoma Saccardo: Distribution, secondary metabolite production and biotechnological applications. Crit Rev Microbiol 35(3):182–196

    Article  CAS  PubMed  Google Scholar 

  • Raman NM, Ramasamy S (2017) Genetic validation and spectroscopic detailing of DHN-melanin extracted from an environmental fungus. Biochem Biophys Rep 12:98–107

    PubMed  PubMed Central  Google Scholar 

  • Rizner TL, Wheeler MH (2003) Melanin biosynthesis in the fungus Curvularia lunata (teleomorph: Cochliobolus lunatus). Can J Microbiol 49:110–119

    Article  CAS  Google Scholar 

  • Roy S, Banerjee A, Mawkhlieng B, Misra AK, Pattanayak A, Harish GD, Singh SK, Ngachan SV, Bansal KC (2015) Genetic diversity and population structure in aromatic and quality rice (Oryza sativa L.) landraces from North-Eastern India. PLoS ONE 10:e0129607. DOI: https://doi.org/10.1371/journal.pone.0129607

  • Selvakumar P, Rajasekar S, Periasamy K, Raaman N (2008) Isolation and characterization of melanin pigment from Pleurotus cystidiosus (telomorph of Antromycopsis macrocarpa). World J Microbiol Biotechnol 24(10):2125–2131

    Article  CAS  Google Scholar 

  • Singha KM, Singh B, Pandey P (2021) Host specifc endophytic microbiome diversity and associated functions in three varieties of scented black rice are dependent on growth stage. Sci Rep 11:12259. https://doi.org/10.1038/s41598-021-91452-4

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Suryanarayanan TS, Ravishankar JP, Venkatesan G, Murali TS (2004) Characterization of the melanin pigment of a cosmopolitan fungal endophyte. Mycol Res 108(8):974–978

    Article  CAS  PubMed  Google Scholar 

  • Suwannarach N, Kumla J, Watanabe B, Matsui K, Lumyong S (2019) Characterization of melanin and optimal conditions for pigment production by an endophytic fungus, Spissiomyces endophytica SDBR-CMU319. PLoS ONE 14(9):e0222187. https://doi.org/10.1371/journal.pone.0222187

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Tamura K, Dudley J, Nei M, Kumar S (2007) MEGA4: molecular evolutionary genetics analysis (MEGA) software version 4.0. Mol Biol Evol 24:1596–1599

    Article  CAS  PubMed  Google Scholar 

  • Toledo AV, Franco MEE, Lopez SMY, Troncozo MI, Saparrat MCN, Balatti PA (2017) Melanins in fungi: types, localization and putative biological roles. Physiol Mol Plant Pathol 99:2–6

    Article  CAS  Google Scholar 

  • Uzma F, Konappa NM, Chowdappa S (2016) Diversity and extracellular enzyme activities of fungal endophytes isolated from medicinal plants of Western Ghats, Karnataka. Egyp J Basi Appl Sci 3:335–342

    Google Scholar 

  • Vergara C, Araujo KEC, Alves LS, Souza SR, Santos LA, Santa-Catarina C, Silva KD, Pereira GMD, Xavier GR, Zilli JE (2018) Contribution of dark septate fungi to the nutrient uptake and growth of rice plants. Braz J Microbiol 49:67–78

    Article  CAS  PubMed  Google Scholar 

  • Wang JJ, Wu CC, Lee CL, Hsieh SL, Chen JB, Lee CI (2017) Antimelanogenic, antioxidant and antiproliferative effects of Antrodia camphorata fruiting bodies on B16–F0 melanoma cells. PLoS ONE 12(1):e0170924. https://doi.org/10.1371/journal.pone.0170924

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • White TJ, Bruns T, Lee S, Taylor J (1990) Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. In: Innis MA, Gelfand DH, Sninsky JJ, White TJ (eds) PCR protocols: a guide to methods and applications. Academic Press, San Diego, pp 315–322

    Google Scholar 

  • Wong SSW, Rani M, Dodagatta-Marri E, Ibrahim-Granet O, Kishore U, Bayry J, Latgé JP, Sahu A, Madan T, Aimanianda V (2018) Fungal melanin stimulates surfactant protein D–mediated opsonization of and host immune response to Aspergillus fumigates spores. J Biol Chem 293(13):4901–4912

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Xu C, Chen T, Li J, Ye M (2020) The structural analysis of melanin and its hepatoprotective effect from Lachnum sp. Proc Biochem 90:249–256

    Article  CAS  Google Scholar 

  • Yu X, Huo L, Liu H, Chen L, Wang Y, Zhu X (2015) Melanin is required for the formation of the multi-cellular conidia in the endophytic fungus Pestalotiopsis microspora. Microbiol Res 179:1–11

    Article  CAS  PubMed  Google Scholar 

  • Zhan F, He Y, Zu Y, Li T, Zhao Z (2011) Characterization of melanin isolated from a dark septate endophyte (DSE), Exophiala pisciphila. World J Microbiol Biotechnol 27:2483–2489

    Article  CAS  Google Scholar 

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Acknowledgements

The authors (KS, RRP) thank ICAR-NBAIM-AMAAS Coordination Unit (ICAR/NBAIM/AMAAS/2017-20/GF/1a/531), Government of India, for providing the financial support of this work. We also appreciate the support of Sophisticated Analytical Instrument Facility (SAIF), Indian Institute of Technology, Madras, and Central Leather Research Institute, Chennai, for spectral analyses.

Funding

This work was supported by Indian Council of Agricultural Research (ICAR), New Delhi, India (Grand No: ICAR-NBAIM-AMAAS/2017-20/GF/1a/531).

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Authors and Affiliations

  1. Department of Biotechnology, JJ College of Arts and Science (Autonomous), Tamil Nadu, Pudukkottai, 622 422, India

    Kannaiah Surendirakumar

  2. Department of Life Sciences, Manipur University, Canchipur, Imphal, 795 003, Manipur, India

    Kannaiah Surendirakumar, Radha Raman Pandey, Surbala Loushambam & Amit Seth

  3. Department of Botany, Bharathiar University, Tamil Nadu, Coimbatore, 641 046, India

    Thangavelu Muthukumar

  4. Centre for Advanced Studies in Botany, University of Madras, Tamil Nadu, Chennai, 600 025, India

    Anbazhagan Sathiyaseelan

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  1. Kannaiah Surendirakumar
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Surendirakumar, K., Pandey, R.R., Muthukumar, T. et al. Characterization and biological activities of melanin pigment from root endophytic fungus, Phoma sp. RDSE17. Arch Microbiol 204, 171 (2022). https://doi.org/10.1007/s00203-022-02788-y

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