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Characterization of pyomelanin secreted by Shewanella sp. and their application in metal recovery

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Abstract

Melanin is a biopolymer with versatile structural and functional properties and diverse applications in recovering toxic chemicals from water and wastewater, biomedical imaging, and as theragnostic agent. We report the structural characterization and biosynthetic pathway of an extracellular pyomelanin secreted by a sponge-associated bacterium, Shewanella sp. (Shewanella-melanin), and their potential application in metal recovery from liquid. Pyomelanin particles of > 50 µm size were found in the culture medium within 48 h of growth, which were formed through the self-polymerization of benzoquinone molecule produced through homogentisic acid pathway. The aspC and hppD genes involved in the biosynthetic pathway of pyomelanin were detected in the whole genome sequence of Shewanella sp. The FT-IR spectra of Shewanella-melanin, at ~ 3300–3420 cm−1 corresponding to the stretching vibration of –NH and –OH, was in good agreement with that of Sepia melanin, while its elemental composition (C/N/H/S of 29.2:8.23:6.41:1.58) was unique. Shewanella-melanin showed ~ 300 and ~ 950 times increased chelation of manganese and iron from a liquid medium supplemented with 2 mM of MnSO4 and FeSO4, respectively, compared to a control. The FT-IR spectrum showed the binding of metal ions to the carboxylic acid, hydroxyl, and amine groups of Shewanella-melanin. The Shewanella-melanin, with its excellent metal biosorption, could be a potential candidate for removing toxic compounds from water, in turn contributing to the fulfillment of sustainable development goal (SDG) 6.

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The data used in the current study are not given in any preprint servers prior to submission to ESPR. The data will be made available upon request.

References

  • Andrews S (2010) FastQC: a quality control tool for high throughput sequence data. Babraham Bioinformatics, Babraham Institute, Cambridge

    Google Scholar 

  • Annamalai J, Ummalyma SB, Pandey A, Bhaskar T (2021) Recent trends in microbial nanoparticle synthesis and potential application in environmental technology: a comprehensive review. Environ Sci Pollut Res 28:49362–49382

    Article  CAS  Google Scholar 

  • Baraldi P, Capelletti R, Crippa P, Romeo N (1979) Electrical characteristics and electret behavior of melanin. J Electrochem Soc 126:1207

    Article  CAS  Google Scholar 

  • Bernsmann F, Ersen O, Voegel J-C, Jan E, Kotov NA, Ball V (2010) Melanin-containing films: Growth from dopamine solutions versus layer-by-layer deposition. ChemPhysChem 11:3299–3305

  • Bolger AM, Lohse M, Usadel B (2014) Trimmomatic: a flexible trimmer for Illumina sequence data. Bioinformatics 30:2114–2120

    Article  CAS  Google Scholar 

  • Bosi E, Donati B, Galardini M, Brunetti S, Sagot M-F, Lió P, Crescenzi P, Fani R, Fondi M (2015) MeDuSa: a multi-draft based scaffolder. Bioinformatics 31:2443–2451

    Article  CAS  Google Scholar 

  • Bowness J, Morton R (1952) Distribution of copper and zinc in the eyes of fresh-water fishes and frogs. Occurrence of metals in melanin fractions from eye tissues. Biochem J 51:530–535

    Article  CAS  Google Scholar 

  • Carletti G, Nervo G, Cattivelli L (2014) Flavonoids and melanins: a common strategy across two kingdoms. Int J Biol Sci 10:1159

    Article  CAS  Google Scholar 

  • Centeno SA, Shamir J (2008) Surface enhanced Raman scattering (SERS) and FTIR characterization of the sepia melanin pigment used in works of art. J Mol Struct 873:149–159

    Article  CAS  Google Scholar 

  • Chedekel Miles R, Ahene Ago B, Lisa Z (1992) Melanin standard method: empirical formula 2. Pigment Cell Res 5:240–246

    Article  Google Scholar 

  • Coon SL, Kotob S, Jarvis BB, Wang S, Fuqua W, Weiner RM (1994) Homogentisic acid is the product of MelA, which mediates melanogenesis in the marine bacterium Shewanella colwelliana D. Appl Environ Microbiol 60:3006–3010

    Article  CAS  Google Scholar 

  • Cordero RJ, Casadevall A (2020) Melanin. Curr Biol 30:R142–R143

    Article  CAS  Google Scholar 

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

    Article  Google Scholar 

  • Galeb HA, Wilkinson EL, Stowell AF, Lin H, Murphy ST, Martin-Hirsch PL, Mort RL, Taylor AM, Hardy JG (2021) Melanins as sustainable resources for advanced biotechnological applications. Global Chall 5:2000102

    Article  Google Scholar 

  • Hong L, Liu Y, Simon JD (2004) Binding of metal ions to melanin and their effects on the aerobic reactivity. Photochem Photobiol 80:477–481

    Article  CAS  Google Scholar 

  • Hong L, Simon JD (2007) Current understanding of the binding sites, capacity, affinity, and biological significance of metals in melanin. J Phys Chem B 111:7938–7947

    Article  CAS  Google Scholar 

  • Jolley KA, Bray JE, Maiden MC (2018) Open-access bacterial population genomics: BIGSdb software, the PubMLST.org website and their applications. Wellcome open research 3

  • Krueger F, James F, Ewels P, Afyounian E, Schuster-Boeckler B (2021) FelixKrueger/TrimGalore: v0 6.7-DOI via Zenodo. Zenodo https://zenodo.org/record/5127899. Accessed 23 Feb 2022

  • Li C, Ji C, Tang B (2018) Purification, characterisation, and biological activity of melanin from Streptomyces sp. FEMS Microbiol Lett 365:fny077

    Article  CAS  Google Scholar 

  • Lindroos M, Hörnström D, Larsson G, Gustavsson M, van Maris AJA (2019) Continuous removal of the model pharmaceutical chloroquine from water using melanin-covered Escherichia coli in a membrane bioreactor. J Hazard Mater 365:74–80

    Article  CAS  Google Scholar 

  • Longo DL, Stefania R, Aime S, Oraevsky A (2017) Melanin-based contrast agents for biomedical optoacoustic imaging and theranostic applications. Int J Mol Sci 18:1719

    Article  Google Scholar 

  • Loring DH, Rantala RTT (1992) Manual for the geochemical analyses of marine sediments and suspended particulate matter. Earth Sci Rev 32:235–283

    Article  CAS  Google Scholar 

  • Lorquin F, Ziarelli F, Amouric A, Di Giorgio C, Robin M, Piccerelle P, Lorquin J (2021) Production and properties of non-cytotoxic pyomelanin by laccase and comparison to bacterial and synthetic pigments. Sci Rep 11:1–16

    Article  Google Scholar 

  • Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–275

    Article  CAS  Google Scholar 

  • Maher S, Mahmoud M, Rizk M, Kalil H (2020) Synthetic melanin nanoparticles as peroxynitrite scavengers, photothermal anticancer and heavy metals removal platforms. Environ Sci Pollut Res 27:19115–19126

    Article  CAS  Google Scholar 

  • Manirethan V, Gupta N, Balakrishnan RM, Raval K (2020a) Batch and continuous studies on the removal of heavy metals from aqueous solution using biosynthesised melanin-coated PVDF membranes. Environ Sci Pollut Res 27:24723–24737

    Article  CAS  Google Scholar 

  • Manirethan V, Raval K, Balakrishnan RM (2020b) Adsorptive removal of trivalent and pentavalent arsenic from aqueous solutions using iron and copper impregnated melanin extracted from the marine bacterium Pseudomonas stutzeri. Environ Pollut 257:113576

    Article  CAS  Google Scholar 

  • McLean J, Purvis O, Williamson B, Bailey E (1998) Role for lichen melanins in uranium remediation. Nature 391:649–650

    Article  CAS  Google Scholar 

  • Mekala LP, Mohammed M, Chinthalapati S, Chinthalapati VR (2019) Pyomelanin production: insights into the incomplete aerobic l-phenylalanine catabolism of a photosynthetic bacterium, Rubrivivax benzoatilyticus JA2. Int J Biol Macromol 126:755–764

    Article  CAS  Google Scholar 

  • Oh J-J, Kim JY, Kim YJ, Kim S, Kim G-H (2021) Utilization of extracellular fungal melanin as an eco-friendly biosorbent for treatment of metal-contaminated effluents. Chemosphere 272:129884

    Article  CAS  Google Scholar 

  • Parvulescu MJS, Martin KL, Mogilevsky P, Patel TA, Street DP, Gupta MK, Hung C-S, Dickerson MB (2021) Biomorphic ceramics: synthesis and characterization of preceramic polymer-modified melaniN. ACS Biomater Sci Eng 7:3103–3113

    Article  CAS  Google Scholar 

  • Pavan ME, López NI, Pettinari MJ (2020) Melanin biosynthesis in bacteria, regulation and production perspectives. Appl Microbiol Biotechnol 104:1357–1370

    Article  CAS  Google Scholar 

  • Paz M, Flückiger R, Boak A, Kagan H, Gallop PM (1991) Specific detection of quinoproteins by redox-cycling staining. J Biol Chem 266:689–692

    Article  CAS  Google Scholar 

  • Płonka P, Grabacka M (2006) Melanin synthesis in microorganisms: biotechnological and medical aspects. Acta Biochim Pol 53:429–443

  • Prota G (1988) Progress in the chemistry of melanins and related metabolites. Med Res Rev 8:525–556

    Article  CAS  Google Scholar 

  • Reali M, Gouda A, Bellemare J, Ménard D, Nunzi J-M, Soavi F, Santato C (2020) Electronic transport in the biopigment sepia melanin. ACS Appl Bio Mater 3:5244–5252

    Article  CAS  Google Scholar 

  • Rodríguez-Rojas A, Mena A, Martín S, Borrell N, Oliver A, Blazquez J (2009) Inactivation of the hmgA gene of Pseudomonas aeruginosa leads to pyomelanin hyperproduction, stress resistance and increased persistence in chronic lung infection. Microbiology 155:1050–1057

    Article  Google Scholar 

  • Sajjan SS, Anjaneya O, Kulkarni GB, Nayak AS, Mashetty SB, Karegoudar T (2013) Properties and functions of melanin pigment from Klebsiella sp. GSK. Microbiol Biotechnol Lett 41:60–69

    Article  CAS  Google Scholar 

  • Sambrook J, Russel DW (2001) Molecular cloning: a laboratory mannual, 3rd edn. Cold Spring Harbour Laboratory, New York

    Google Scholar 

  • Seelam SD, Agsar D, Halmuthur MSK, Reddy Shetty P, Vemireddy S, Reddy KM, Umesh MK, Rajitha CH (2021) Characterization and photoprotective potentiality of lime dwelling Pseudomonas mediated melanin as sunscreen agent against UV-B radiations. J Photochem Photobiol B 216:112126

    Article  CAS  Google Scholar 

  • Seemann T (2014) Prokka: rapid prokaryotic genome annotation. Bioinformatics 30:2068–2069

    Article  CAS  Google Scholar 

  • Shin S, Kwak H, Hyun J (2018) Melanin nanoparticle-incorporated silk fibroin hydrogels for the enhancement of printing resolution in 3D-Projection stereolithography of poly(ethylene glycol)-tetraacrylate bio-ink. ACS Appl Mater Interfaces 10:23573–23582

    Article  CAS  Google Scholar 

  • Silveira Maranhão Fd, Gomes F, Thode S, Das DB, Pereira E, Lima N, Carvalho F, Aboelkheir M, Costa V, Pal K (2021) Oil spill sorber based on extrinsically magnetizable porous geopolymer. Materials 14:5641

    Article  Google Scholar 

  • Singh D, Kumar J, Kumar A (2018) Isolation of pyomelanin from bacteria and evidences showing its synthesis by 4-hydroxyphenylpyruvate dioxygenase enzyme encoded by hppD gene. Int J Biol Macromol 119:864–873

    Article  CAS  Google Scholar 

  • Singh S, Nimse SB, Mathew DE, Dhimmar A, Sahastrabudhe H, Gajjar A, Ghadge VA, Kumar P, Shinde PB (2021) Microbial melanin: Recent advances in biosynthesis, extraction, characterization, and applications. Biotechnol Adv 53:107773

  • Siwicka ZE, Son FA, Battistella C, Moore MH, Korpanty J, McCallum NC, Wang Z, Johnson BJ, Farha OK, Gianneschi NC (2021) Synthetic porous melanin. J Am Chem Soc 143:3094–3103

    Article  CAS  Google Scholar 

  • Sono K, Lye D, Moore CA, Boyd WC, Gorlin TA, Belitsky JM (2012): Melanin-based coatings as lead-binding agents. Bioinorganic chemistry and applications 361803

  • Steinert M, Flugel M, Schuppler M, Helbig JM, Supriyono A, Proksch P, Luck PC (2001) The Lly protein is essential for p-hydroxyphenyl pyruvate dioxygenase activity in Legionella pneumophila. FEMS Microbiol Lett 203:41–47

  • Takeda K, Takahashi N-H, Shibahara S (2007) Neuroendocrine functions of melanocytes: beyond the skin-deep melanin maker. Tohoku J Exp Med 211:201–221

    Article  CAS  Google Scholar 

  • Tarangini K, Mishra S (2014) Production of melanin by soil microbial isolate on fruit waste extract: two step optimization of key parameters. Biotechnol Rep 4:139–146

    Article  Google Scholar 

  • Tran-Ly AN, Reyes C, Schwarze FWMR, Ribera J (2020) Microbial production of melanin and its various applications. World J Microbiol Biotechnol 36:170

    Article  CAS  Google Scholar 

  • Turick CE, Caccavo F Jr, Tisa LS (2003) Electron transfer from Shewanella algae BrY to hydrous ferric oxide is mediated by cell-associated melanin. FEMS Microbiol Lett 220:99–104

    Article  CAS  Google Scholar 

  • Turick CE, Caccavo F, Tisa LS (2008) Pyomelanin is produced by Shewanella algae BrY and affected by exogenous iron. Can J Microbiol 54:334–339

    Article  CAS  Google Scholar 

  • Turick CE, Knox AS, Becnel JM, Ekechukwu AA, Milliken CE (2010) Properties and function of pyomelanin. Biopolymers 449:72

    Google Scholar 

  • Vijayan V, Jasmin C, Anas A, Kuttan SP, Vinothkumar S, Subrayan PP, Nair S (2017) Sponge-associated bacteria produce non-cytotoxic melanin which protects animal cells from photo-toxicity. Appl Biochem Biotechnol 183:396–411

    Article  CAS  Google Scholar 

  • Wang Z, Tschirhart T, Schultzhaus Z, Kelly Erin E, Chen A, Oh E, Nag O, Glaser Evan R, Kim E, Lloyd Pamela F, Charles Paul T, Li W, Leary D, Compton J, Phillips Daniel A, Dhinojwala A, Payne Gregory F, Vora Gary J, Johnson Karyn N (2020) Melanin produced by the fast-growing marine bacterium Vibrio natriegens through heterologous biosynthesis: characterization and application. Appl Environ Microbiol 86:e02749-e2819

    CAS  Google Scholar 

  • Wick RR, Judd LM, Gorrie CL, Holt KE (2017) Unicycler: resolving bacterial genome assemblies from short and long sequencing reads. PLoS Comput Biol 13:e1005595

    Article  Google Scholar 

  • Xie W, Pakdel E, Liang Y, Kim YJ, Liu D, Sun L, Wang X (2019) Natural Eumelanin and its derivatives as multifunctional materials for bioinspired applications: a review. Biomacromolecules 20:4312–4331

    Article  CAS  Google Scholar 

  • Zeng Z, Guo X-P, Cai X, Wang P, Li B, Yang J-L, Wang X (2017) Pyomelanin from Pseudoalteromonas lipolytica reduces biofouling. Microb Biotechnol 10:1718–1731

    Article  CAS  Google Scholar 

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Acknowledgements

The authors thank the Director, CSIR-National Institute of Oceanography, Goa, and Scientist-in-Charge CSIR-NIO Regional Centre Kochi for extending all the required support. This is CSIR-NIO contribution 6962.

Funding

This work was implemented with the financial support of the Council of Scientific and Industrial Research, Govt of India (CSIR-NIO/FTT/2016/PME-1/7, MLP1603 and BIOPROSmar (MLP2019)). SPK received fellowship from a research project supported by Department of Biotechnology, Govt of India (BT/PR26858/AAQ/3/880/2017).

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Authors and Affiliations

  1. CSIR-National Institute of Oceanography, Regional Centre Kochi, Dr. Salim Ali Road, Kochi, Kerala, 682 018, India

    Sreelakshmi Parakkaparambil Kuttan, Anas Abdulaziz, Jasmin Chekidhenkuzhiyil & Devika Raj

  2. School of Environmental Sciences, Mahatma Gandhi University, Priyadarsini Hills, Kottayam, Kerala, 686 560, India

    Mahesh Mohan

  3. Department of Chemistry, National Institute of Technology Calicut, Kozhikode, Kerala, 673 601, India

    Sujith Athiyanathil

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  1. Sreelakshmi Parakkaparambil Kuttan
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  2. Anas Abdulaziz
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Contributions

The design of the work and the preparation of manuscript were done by AA. SPK, JC, SA, KRD, and MM contributed to the experimental data and analysis. KRD was involved in the analysis of whole-genome data. MM and SA were involved in the manuscript preparation.

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Correspondence to Anas Abdulaziz.

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Kuttan, S.P., Abdulaziz, A., Chekidhenkuzhiyil, J. et al. Characterization of pyomelanin secreted by Shewanella sp. and their application in metal recovery. Environ Sci Pollut Res 30, 6705–6715 (2023). https://doi.org/10.1007/s11356-022-22686-6

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