Journal of Multidisciplinary Applied Natural Science
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2.1
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an Open Access Journal
2.1
Calculated on 05 May, 2025
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https://doi.org/10.47352/jmans.2774-3047.446
Muhammad Asril
Achmad Gus Fahmi
Annisa Maulidia Rahayyu
Taufik Priya Azzahwa
Dian Anggria Sari
Didik Supriyadi
Microbial melanin has attracted increasing attention due to its multifunctional properties, particularly its antioxidant activity, making it attractive for biotechnological applications. Volcanic soils represent extreme, metal-rich ecosystems that may harbor melanogenic bacteria with unique metabolic adaptations. This study aimed to isolate and characterize melanin-producing bacteria from the volcanic soil of Mount Anak Krakatau, evaluating their melanogenic capacity, antioxidant activity, and volatile metabolite profiles. Eight bacterial isolates were obtained using tyrosine agar supplemented with CuSO₄ to selectively promote tyrosinase-mediated melanogenesis. Three isolates (AK3, AK5, and AK7) exhibited clear melanogenic phenotypes, as indicated by the progressive development of brown to black pigmentation during cultivation. Tyrosinase activity and L-DOPA concentration showed dynamic changes throughout the production period, with isolate AK5 demonstrating the highest tyrosinase activity (4,500 ± 135.27 U/mL) and L-DOPA accumulation (up to 89.82 ± 0.13 µg/mL), reflecting efficient biotransformation of L-tyrosine. The antioxidant activity of free-cell supernatants, evaluated using the ferric reducing antioxidant power (FRAP) and total antioxidant capacity (TAC) assays, yielded substantial values, with AK5 exhibiting the highest performance among the testing isolates and further identified as Bacillus cereus AK5 by phylogenetic analysis of 16S rRNA gene sequences. Gas chromatography–mass spectrometry (GC–MS) analysis revealed distinct, isolate-specific volatile organic compound (VOC) profiles, which we hypothesize may reflect adaptive secondary metabolic responses to oxidative stress generated during melanogenesis, though this link requires further experimental validation. To the best of our knowledge and based on the available literature, this study reports the first isolation of melanin-producing bacteria with antioxidant potential from volcanic soil at Mount Anak Krakatau, highlighting this environment as a promising source of melanogenic bacteria for pharmaceutical applications.
[1] O. Kraseasintra, S. Sensupa, K. Mahanil, S. Yoosathaporn, J. Pekkoh, S. Srinuanpan, W. Pathom-Aree, and C. Pumas. (2023). "Optimization of Melanin Production by Streptomyces antibioticus NRRL B-1701 Using Arthrospira (Spirulina) platensis Residues Hydrolysates as Low-Cost L-Tyrosine Supplement". BioTech. 12 (1). 10.3390/biotech12010024.
DOI: https://doi.org/10.3390/biotech12010024[2] X. Wan, H. Liu, Y. Liao, Y. Su, J. Geng, M. Yang, X. Chen, and P. Shen. (2007). "Isolation of a Novel Strain of Aeromonas media Producing High Levels of DOPA-Melanin and Assessment of the Photoprotective Role of the Melanin in Bioinsecticide Applications". Journal of Applied Microbiology. 103 (6): 2533-2541. 10.1111/j.1365-2672.2007.03502.x.
DOI: https://doi.org/10.1111/j.1365-2672.2007.03502.x[3] S. Singh, S. B. Nimse, D. E. Mathew, A. Dhimmar, H. Sahastrabudhe, A. Gajjar, V. A. Ghadge, P. Kumar, and P. B. Shinde. (2021). "Microbial Melanin: Recent Advances in Biosynthesis, Extraction, Characterization, and Applications". Biotechnology Advances. 53 : 107773. 10.1016/j.biotechadv.2021.107773.
DOI: https://doi.org/10.1016/j.biotechadv.2021.107773[4] N. E. A. El-Naggar and W. I. A. Saber. (2022). "Natural Melanin: Current Trends and Future Approaches, with Especial Reference to Microbial Source". Polymers. 14 (7): 1339. 10.3390/polym14071339.
DOI: https://doi.org/10.3390/polym14071339[5] P. Sharma, T. A. Singh, B. Bharat, S. Bhasin, and H. A. Modi. (2018). "Approach Towards Different Fermentative Techniques for the Production of Bioactive Actinobacterial Melanin". Beni-Suef University Journal of Basic and Applied Sciences. 7 (4): 695-700. 10.1016/j.bjbas.2018.08.002.
DOI: https://doi.org/10.1016/j.bjbas.2018.08.002[6] I. Pralea, R. Moldovan, A. Petrache, M. Ilies, S. Heghes, I. Ielciu, R. Nicoara, M. Moldovan, M. Ene, M. Radu, A. Uifalean, and C. Iuga. (2019). "From Extraction to Advanced Analytical Methods: The Challenges of Melanin Analysis". International Journal of Molecular Sciences. 20 (16): 3943. 10.3390/ijms20163943.
DOI: https://doi.org/10.3390/ijms20163943[7] M. Asril, R. I. Astuti, I. Rusmana, and A. T. Wahyudi. (2025). "Improved Eumelanin Production, Phenolic Content, Flavonoid Content, and Antioxidant Activity by Streptomyces lasalocidi NTB 42 Following Copper Sulfate Supplementation". Biocatalysis and Agricultural Biotechnology. 66 : 103602. 10.1016/j.bcab.2025.103602.
DOI: https://doi.org/10.1016/j.bcab.2025.103602[8] J. Guo, Z. Rao, T. Yang, Z. Man, M. Xu, and X. Zhang. (2014). "High-Level Production of Melanin by a Novel Isolate of Streptomyces kathirae". FEMS Microbiology Letters. 357 (1): 85-91. 10.1111/1574-6968.12497.
DOI: https://doi.org/10.1111/1574-6968.12497[9] N. E. A. El-Naggar and S. M. El-Ewasy. (2017). "Bioproduction, Characterization, Anticancer and Antioxidant Activities of Extracellular Melanin Pigment Produced by Newly Isolated Microbial Cell Factories Streptomyces glaucescens NEAE-H". Scientific Reports. 7 : 42129. 10.1038/srep42129.
DOI: https://doi.org/10.1038/srep42129[10] A. S. Gamal Shalaby, T. I. M. Ragab, M. M. I. Helal, and M. A. Esawy. (2019). "Optimization of Bacillus licheniformis MAL Tyrosinase: In Vitro Anticancer Activity for Brown and Black Eumelanin". Heliyon. 5 (5): e01657. 10.1016/j.heliyon.2019.e01657.
DOI: https://doi.org/10.1016/j.heliyon.2019.e01657[11] V. Ghadge, P. Kumar, S. Singh, D. E. Mathew, S. Bhattacharya, S. B. Nimse, and P. B. Shinde. (2020). "Natural Melanin Produced by the Endophytic Bacillus subtilis 4NP-BL Associated with the Halophyte Salicornia brachiata". Journal of Agricultural and Food Chemistry. 68 (25): 6854-6863. 10.1021/acs.jafc.0c01997.
DOI: https://doi.org/10.1021/acs.jafc.0c01997[12] O. F. Restaino, M. Scognamiglio, S. F. Mirpoor, M. Cammarota, R. Ventriglia, C. V. L. Giosafatto, A. Fiorentino, R. Porta, and C. Schiraldi. (2022). "Enhanced Streptomyces roseochromogenes Melanin Production by Using the Marine Renewable Source Posidonia oceanica Egagropili". Applied Microbiology and Biotechnology. 106 (21): 7265-7283. 10.1007/s00253-022-12191-8.
DOI: https://doi.org/10.1007/s00253-022-12191-8[13] O. F. Restaino, T. Kordjazi, F. Tancredi, P. Manini, F. Lanzillo, F. Raganati, A. Marzocchella, R. Porta, and L. Mariniello. (2025). "Metal Ion Supplementation to Boost Melanin Production by Streptomyces nashvillensis". International Journal of Molecular Sciences. 26 (1): 416. 10.3390/ijms26010416.
DOI: https://doi.org/10.3390/ijms26010416[14] D. Fiantis, F. I. Ginting, Gusnidar, M. Nelson, E. Van Ranst, and B. Minasny. (2021). "Geochemical Characterization and Evolution of Soils from Krakatau Islands". Eurasian Soil Science. 54 (11): 1629-1643. 10.1134/S1064229321110077.
DOI: https://doi.org/10.1134/S1064229321110077[15] A. R. Setiawati, J. Lumbanraja, A. I. Kurnia, M. Hidayat, S. N. Aini, and D. Prasetyo. (2023). "Changes of Soil Chemistry Characteristics of Tephra Mount Anak Krakatau, Indonesia, Through Leaching Experiment". Journal of Tropical Soils. 28 (2): 57-70. 10.5400/jts.2023.v28i2.57-70.
DOI: https://doi.org/10.5400/jts.2023.v28i2.57-70[16] M. A. Burmasova, A. A. Utebaeva, E. V. Sysoeva, and M. A. Sysoeva. (2019). "Melanins of Inonotus obliquus: Bifidogenic and Antioxidant Properties". Biomolecules. 9 (6): 248. 10.3390/biom9060248.
DOI: https://doi.org/10.3390/biom9060248[17] N. P. Arslan, F. Azad, T. Orak, A. Budak-Savas, S. Ortucu, P. Dawar, M. O. Baltaci, H. Ozkan, N. Esim, and M. Taskin. (2025). "A Review on Bacteria-Derived Antioxidant Metabolites: Their Production, Purification, Characterization, Potential Applications, and Limitations". Archives of Pharmacal Research. 48 (4): 253-292. 10.1007/s12272-025-01541-5.
DOI: https://doi.org/10.1007/s12272-025-01541-5[18] S. Schulz and J. S. Dickschat. (2007). "Bacterial Volatiles: The Smell of Small Organisms". Natural Product Reports. 24 (4): 814-842. 10.1039/B507392H.
DOI: https://doi.org/10.1039/b507392h[19] C. N. Kanchiswamy, M. Malnoy, and M. E. Maffei. (2015). "Chemical Diversity of Microbial Volatiles and Their Potential for Plant Growth and Productivity". Frontiers in Plant Science. 6 : 151. 10.3389/fpls.2015.00151.
DOI: https://doi.org/10.3389/fpls.2015.00151[20] M. I. Mhlongo, L. A. Piater, and I. A. Dubery. (2022). "Profiling of Volatile Organic Compounds from Four Plant Growth-Promoting Rhizobacteria by SPME-GC-MS: A Metabolomics Study". Metabolites. 12 (8): 763. 10.3390/metabo12080763.
DOI: https://doi.org/10.3390/metabo12080763[21] M. Asril, R. I. Astuti, I. Rusmana, N. P. R. A. Krishanti, and A. T. Wahyudi. (2024). "Characterization and Antioxidant Activity of Eumelanin Produced by Streptomyces lasalocidi NTB 42". Biocatalysis and Agricultural Biotechnology. 61 : 103361. 10.1016/j.bcab.2024.103361.
DOI: https://doi.org/10.1016/j.bcab.2024.103361[22] K. M. Raval, P. S. Vaswani, and D. R. Majumder. (2012). "Biotransformation of a Single Amino-Acid L-Tyrosine into a Bioactive Molecule L-DOPA". International Journal of Scientific and Research Publications. 2 (5): 1-9.
[23] A. I. El-Batal, G. S. El-Sayyad, A. El-Ghamery, and M. Gobara. (2017). "Response Surface Methodology Optimization of Melanin Production by Streptomyces cyaneus and Synthesis of Copper Oxide Nanoparticles Using Gamma Radiation". Journal of Cluster Science. 28 (3): 1083-1112. 10.1007/s10876-016-1101-0.
DOI: https://doi.org/10.1007/s10876-016-1101-0[24] T. Chen, S. Liou, H. Wu, F. Tsai, C. Tsai, C. Huang, and Y. Chang. (2010). "New Analytical Method for Investigating the Antioxidant Power of Food Extracts on the Basis of Their Electron-Donating Ability: Comparison to the Ferric Reducing Antioxidant Power (FRAP) Assay". Journal of Agricultural and Food Chemistry. 58 (15): 8477-8480. 10.1021/jf9044292.
DOI: https://doi.org/10.1021/jf9044292[25] P. Prieto, M. Pineda, and M. Aguilar. (1999). "Spectrophotometric Quantitation of Antioxidant Capacity Through the Formation of a Phosphomolybdenum Complex: Specific Application to the Determination of Vitamin E". Analytical Biochemistry. 269 (2): 337-341. 10.1006/abio.1999.4019.
DOI: https://doi.org/10.1006/abio.1999.4019[26] F. Ditterich, C. Poll, G. J. Pronk, K. Heister, A. Chandran, T. Rennert, I. Kogel-Knabner, and E. Kandeler. (2016). "Succession of Soil Microbial Communities and Enzyme Activities in Artificial Soils". Pedobiologia - Journal of Soil Ecology. 59 (3): 93-104. 10.1016/j.pedobi.2016.03.002.
DOI: https://doi.org/10.1016/j.pedobi.2016.03.002[27] A. Neaman, J. Schoffer, C. Navarro-Villarroel, C. Pelosi, P. Penaloza, E. Dovletyarova, and J. Schneider. (2024). "Copper Contamination in Agricultural Soils: A Review of the Effects of Climate, Soil Properties, and Prolonged Copper Pesticide Application in Vineyards and Orchards". Plant, Soil and Environment. 70 (7): 407-417. 10.17221/501/2023-PSE.
DOI: https://doi.org/10.17221/501/2023-PSE[28] O. F. Restaino, P. Manini, T. Kordjazi, M. L. Alfieri, M. Rippa, L. Mariniello, and R. Porta. (2024). "Biotechnological Production and Characterization of Extracellular Melanin by Streptomyces nashvillensis". Microorganisms. 12 (2): 297. 10.3390/microorganisms12020297.
DOI: https://doi.org/10.3390/microorganisms12020297[29] E. Valipour and B. Arikan. (2016). "Increased Production of Tyrosinase from Bacillus megaterium Strain M36 by the Response Surface Method". Archives of Biological Sciences. 68 (3): 659-668. 10.2298/ABS151002058V.
DOI: https://doi.org/10.2298/ABS151002058V[30] J. Zhang, J. Cai, Y. Deng, Y. Chen, and G. Ren. (2007). "Characterization of Melanin Produced by a Wild-Type Strain of Bacillus cereus". Frontiers of Biology in China. 2 (1): 26-29. 10.1007/s11515-007-0004-8.
DOI: https://doi.org/10.1007/s11515-007-0004-8[31] C. Marín-Sanhueza, A. Echeverría-Vega, A. Gómez, G. Cabrera-Barjas, R. Romero, and A. Banerjee. (2022). "Stress-Dependent Biofilm Formation and Bioactive Melanin Pigment Production by a Thermophilic Bacillus Species from Chilean Hot Spring". Polymers. 14 (4): 680. 10.3390/polym14040680.
DOI: https://doi.org/10.3390/polym14040680[32] B. An, Y. Zhan, Q. Cheng, J. Cai, and X. Gu. (2024). "Production Improvement and Photoprotection of Melanin Produced by Bacillus thuringiensis". BioControl. 69 (2): 157-167. 10.1007/s10526-024-10250-9.
DOI: https://doi.org/10.1007/s10526-024-10250-9[33] N. J. Kang, H. S. Jin, S. E. Lee, H. J. Kim, H. Koh, and D. W. Lee. (2020). "New Approaches Towards the Discovery and Evaluation of Bioactive Peptides from Natural Resources". Critical Reviews in Environmental Science and Technology. 50 (1): 72-103. 10.1080/10643389.2019.1619376.
DOI: https://doi.org/10.1080/10643389.2019.1619376[34] M. Abdul Sattar and A. Patnaik. (2023). "Molecular Insights into Antioxidant Efficiency of Melanin: A Sustainable Antioxidant for Natural Rubber Formulations". The Journal of Physical Chemistry B. 127 (38): 8242-8256. 10.1021/acs.jpcb.3c03523.
DOI: https://doi.org/10.1021/acs.jpcb.3c03523[35] M. Pang, R. Xu, R. Xi, H. Yao, K. Bao, R. Peng, H. Zhi, K. Zhang, R. He, Y. Su, X. Liu, and D. Ming. (2024). "Molecular Understanding of the Therapeutic Potential of Melanin Inhibiting Natural Products". RSC Medicinal Chemistry. 15 (7): 2226-2253. 10.1039/D4MD00224E.
DOI: https://doi.org/10.1039/D4MD00224E[36] Z. Li, H. Heng, Q. Qin, L. Chen, Y. Wang, and Z. Zhou. (2022). "Physicochemical Properties, Molecular Structure, Antioxidant Activity, and Biological Function of Extracellular Melanin from Ascosphaera apis". Journal of Zhejiang University Science B. 23 (5): 365-381. 10.1631/jzus.B2100718.
DOI: https://doi.org/10.1631/jzus.B2100718[37] M. Kuczynska, P. Jakubek, and A. Bartoszek. (2022). "More than Just Antioxidants: Redox-Active Components and Mechanisms Shaping Redox Signalling Network". Antioxidants (Basel). 11 (12). 10.3390/antiox11122403.
DOI: https://doi.org/10.3390/antiox11122403[38] H. A. T. Nguyen, T. P. Ho, D. Mangelings, A. Van Eeckhaut, Y. Vander Heyden, and H. T. M. Tran. (2024). "Antioxidant, Neuroprotective, and Neuroblastoma Cells (SH-SY5Y) Differentiation Effects of Melanins and Arginine-Modified Melanins from Daedaleopsis tricolor and Fomes fomentarius". BMC Biotechnology. 24 (1): 89. 10.1186/s12896-024-00918-6.
DOI: https://doi.org/10.1186/s12896-024-00918-6[39] W. Cao, X. Zhou, N. C. McCallum, Z. Hu, Q. Z. Ni, U. Kapoor, C. M. Heil, K. S. Cay, T. Zand, A. J. Mantanona, A. Jayaraman, A. Dhinojwala, D. D. Deheyn, M. D. Shawkey, M. D. Burkart, J. D. Rinehart, and N. C. Gianneschi. (2021). "Unraveling the Structure and Function of Melanin Through Synthesis". Journal of the American Chemical Society. 143 (7): 2622-2637. 10.1021/jacs.0c12322.
DOI: https://doi.org/10.1021/jacs.0c12322[40] A. Tariq, M. Athar, J. Ara, V. Sultana, and M. Ahmad. (2015). "Biochemical Evaluation of Antioxidant Activity in Extracts and Polysaccharide Fractions of Seaweeds". Global Journal of Environmental Science and Management. 1 (1): 47-62. 10.7508/gjesm.2015.01.005.
[41] T. Kordjazi, L. Mariniello, C. V. L. Giosafatto, R. Porta, and O. F. Restaino. (2024). "Streptomycetes as Microbial Cell Factories for the Biotechnological Production of Melanin". International Journal of Molecular Sciences. 25 (5): 3013. 10.3390/ijms25053013.
DOI: https://doi.org/10.3390/ijms25053013[42] B. Ginting, E. Sufriadi, E. Harnelly, N. Isnaini, F. Mulana, I. H. Suparto, A. Ilmiawati, E. Ernawati, S. Muhammad, M. Syakira, and C. D. Riski. (2023). "Identification of Volatile Compounds Contained in the Therapeutic Essential Oils from Pogostemon cablin, Melaleuca leucadendra, and Mentha piperita and Their Purified Fractions". Journal of Advanced Pharmaceutical Technology and Research. 14 (3): 208-212. 10.4103/JAPTR.JAPTR_161_23.
DOI: https://doi.org/10.4103/JAPTR.JAPTR_161_23[43] N. Roy, R. Das, R. Paira, and P. Paira. (2023). "Different Routes for the Construction of Biologically Active Diversely Functionalized Bicyclo[3.3.1]Nonanes: An Exploration of New Perspectives for Anticancer Chemotherapeutics". RSC Advances. 13 (32): 22389-22480. 10.1039/D3RA02003G.
DOI: https://doi.org/10.1039/D3RA02003G[44] C. F. Ajilogba and O. O. Babalola. (2019). "GC-MS Analysis of Volatile Organic Compounds from Bambara Groundnut Rhizobacteria and Their Antibacterial Properties". World Journal of Microbiology and Biotechnology. 35 (6): 83. 10.1007/s11274-019-2660-7.
DOI: https://doi.org/10.1007/s11274-019-2660-7[45] I. C. F. R. Ferreira, P. Baptista, M. Vilas-Boas, and L. Barros. (2007). "Free-Radical Scavenging Capacity and Reducing Power of Wild Edible Mushrooms from Northeast Portugal: Individual Cap and Stipe Activity". Food Chemistry. 100 (4): 1511-1516. 10.1016/j.foodchem.2005.11.043.
DOI: https://doi.org/10.1016/j.foodchem.2005.11.043[46] B. C. S. Santos, A. S. Pires, C. H. Yamamoto, M. R. C. Couri, A. G. Taranto, M. S. Alves, A. L. D. S. De Matos Araújo, and O. V. De Sousa. (2018). "Methyl Chavicol and Its Synthetic Analogue as Possible Antioxidant and Antilipase Agents Based on the In Vitro and In Silico Assays". Oxidative Medicine and Cellular Longevity. 2018 : 2189348. 10.1155/2018/2189348.
DOI: https://doi.org/10.1155/2018/2189348[47] T. Netzker, E. M. F. Shepherdson, M. P. Zambri, and M. A. Elliot. (2020). "Bacterial Volatile Compounds: Functions in Communication, Cooperation, and Competition". Annual Review of Microbiology. 74 : 409-430. 10.1146/annurev-micro-011320-015542.
DOI: https://doi.org/10.1146/annurev-micro-011320-015542[48] S. Moser and H. Pichler. (2019). "Identifying and Engineering the Ideal Microbial Terpenoid Production Host". Applied Microbiology and Biotechnology. 103 (14): 5501-5516. 10.1007/s00253-019-09892-y.
DOI: https://doi.org/10.1007/s00253-019-09892-y[49] Y. Yang, D. Li, R. Wang, X. Ji, Y. Wang, Q. Guo, and T. Shi. (2025). "Recent Advances and Multiple Strategies for the Synthesis of Terpenoid Fragrances and Flavors in Model Microorganisms". Biotechnology Advances. 83 : 108646. 10.1016/j.biotechadv.2025.108646.
DOI: https://doi.org/10.1016/j.biotechadv.2025.108646[50] R. Schmidt, V. Cordovez, W. De Boer, J. Raaijmakers, and P. Garbeva. (2015). "Volatile Affairs in Microbial Interactions". The ISME Journal. 9 (11): 2329-2335. 10.1038/ismej.2015.42.
DOI: https://doi.org/10.1038/ismej.2015.42[51] N. S. A. Hagaggi and U. M. Abdul-Raouf. (2025). "Recent Trends in Microbial Production of Alkanes". World Journal of Microbiology and Biotechnology. 41 (9): 320. 10.1007/s11274-025-04536-y.
DOI: https://doi.org/10.1007/s11274-025-04536-y[52] R. M. Dickey, A. M. Forti, and A. M. Kunjapur. (2021). "Advances in Engineering Microbial Biosynthesis of Aromatic Compounds and Related Compounds". Bioresources and Bioprocessing. 8 (1): 91. 10.1186/s40643-021-00434-x.
DOI: https://doi.org/10.1186/s40643-021-00434-x[53] X. Lv, Y. Li, T. Cui, M. Sun, F. Bai, X. Li, J. Li, and S. Yi. (2020). "Bacterial Community Succession and Volatile Compound Changes During Fermentation of Shrimp Paste from Chinese Jinzhou Region". LWT - Food Science and Technology. 122 : 108998. 10.1016/j.lwt.2019.108998.
DOI: https://doi.org/10.1016/j.lwt.2019.108998[54] D. Plamada, A. S. Nemes, B. E. Teleky, M. S. Pascuta, R. Odocheanu, L. Mitrea, L. F. Calinoiu, K. Szabo, and D. C. Vodnar. (2024). In: "Microbial Production of Food Bioactive Compounds". 1-24. 10.1007/978-3-030-81403-8_53-1.
DOI: https://doi.org/10.1007/978-3-030-81403-8_53-1