Extraction and Characterization of Melanin Pigment from Local Isolated Pseudomonas aeruginosa
DOI:
https://doi.org/10.30526/38.2.3583Keywords:
Melanin pigments, Pseudomonas aeruginosa, Extraction, Purification, UV-vis spectroscopy, FT-IR spectroscopyAbstract
In this present study, a clinically isolated of Pseudomonas spp. from a variety of sources, such as burn patients, wound infections, and UTI patients, that can produce melanin pigment in the presence of 2 % L-tyrosine in both liquid and solid media. These bacteria were identified as Pseudomonas aeruginosa based on morphological, biochemical, and VITEK-2 compact system analysis. The higher melanin producer isolate Ps81, which produces melanin at rate of 3.018933 µg/ml, has been chosen for further investigation steps. The pigment was extracted by several steps of alkali dissolving and acid precipitation followed by washing with organic solvents. Chemical analysis of the pigment particles revealed that they were acid resistant, alkali soluble, insoluble in water, soluble in methanol 100%, ethanol 70% and DMSO but insoluble in ethyl acetate and chloroform. The alkali melanin solution displayed a significant UV absorbance at 273.5 nm and decreased toward visible light and infrared region, the pigments UV-visible spectrum ranged from 200 to 800 nm. FT-IR spectroscopy was used to further characterize the melanin pigment. The infrared spectrum of purified melanin extracted from Ps 81 exhibited a broad absorption band around 3280.82 cm−1 which correspond to the presence of -OH group and N-H groups.
References
1. Ketelboeter L, Potharla VS, Bardy SL. NTBC treatment of the pyomelanogenic Pseudomonas aeruginosa clinical isolate PA1111 inhibits pigment production and increases sensitivity to oxidative stress. Curr Microbiol. 2014;69(3):343–8. https://doi.org/10.1007/s00284-014-0583-8.
2. El-Naggar NEA, El-Ewasy SM. Bioproduction, characterization, anticancer and antioxidant activities of extracellular melanin pigment produced by newly isolated microbial cell factories Streptomyces Glaucescens NEAE-H. Sci Rep. 2017;7:42129. https://doi.org/10.1038/srep42129.
3. Ammanagi AI, Shivasharana, Krishnaveni, Badiger AS, Ramaraj VK, Srinath, Karthik Y. A biotechnological approach to optimization and production of melanin by Brevibacillus invocatus strain IBA, under submerged fermentation. Biomed (Trivandrum). 2022;42(2):318–24. https://doi.org/10.51248/.v42i2.1315.
4. Agunbiade M, Le Roes-Hill M. Application of bacterial tyrosinases in organic synthesis. World J Microbiol Biotechnol. 2022;38(1):2. https://doi.org/10.1007/s11274-021-03200-6.
5. Kamarudheen N, Naushad T, Rao KVB. Biosynthesis, characterization and antagonistic applications of extracellular melanin pigment from marine Nocardiopsis sps. Indian J Pharm Educ. 2019;53(Suppl 2):s112–20. https://doi.org/10.5530/ijper.53.2s.55.
6. Mohammed HA, Zgair AK. Detection of quorum sensing genes of Pseudomonas aeruginosa isolated from different areas in Iraq. Iraqi J Sci. 2022;63(11):4665–73. https://doi.org/10.24996/ijs.2022.63.11.5.
7. Surwase SN, Jadhav SB, Phugare SS, Jadhav JP. Optimization of melanin production by Brevundimonas sp. SGJ using response surface methodology. 3 Biotech. 2013;3(3):187–94. https://doi.org/10.1007/s13205-012-0082-4.
8. Zhang J, Cai J, Deng Y, Chen Y, Ren G. Characterization of melanin produced by a wild-type strain of Bacillus cereus. Front Biol China. 2007;2(1):26–9. https://doi.org/10.1007/s11515-007-0004-8.
9. Noman AE, Al-Barha NS, Chen F. Characterization of physicochemical properties of melanin produced by Gluconobacter oxydans FBFS 97. Fermentation. 2022;8(11):574. https://doi.org/10.3390/fermentation8110574.
10. Sajjan S, Kulkarni G, Yaligara V, Kyoung L, Karegoudar TB. Purification and physiochemical characterization of melanin pigment from Klebsiella sp. GSK. J Microbiol Biotechnol. 2010;20(11):1513–20. https://doi.org/10.4014/jmb.1002.02006.
11. Hossain M, Saha S, Rahman M, Singha J, Mamun A. Isolation, identification and antibiogram study of Pseudomonas aeruginosa from cattle in Bangladesh. J Vet Adv. 2013;3(4):180. https://doi.org/10.5455/jva.20130717123841.
12. Kodaka H, Iwata M, Yumoto S, Kashitani F. Evaluation of a new agar medium containing cetrimide, kanamycin and nalidixic acid for isolation and enhancement of pigment production of Pseudomonas aeruginosa in clinical samples. J Basic Microbiol. 2003;43(5):407–13. https://doi.org/10.1002/jobm.200310264.
13. Sulaiman SD, Abdulhasan GA. Curcumin as efflux pump inhibitor agent for enhancement treatment against multidrug-resistant Pseudomonas aeruginosa isolates. Iraqi J Sci. 2020;61(1):59–67. https://doi.org/10.24996/ijs.2020.61.1.6.
14. Su SS, Lae KZW, Ngwe H. Isolation and identification of Pseudomonas aeruginosa from the clinical soil. Univ Yangon Res J. 2018;8:271–5.
15. Gheni MR, Odaa NH. The antimicrobial activity of melanin-mediated synthesis of silver nanoparticles. Egypt J Hosp Med. 2023;90(1):3383–94. https://doi.org/10.21608/ejhm.2023.291442.
16. Wellinghausen N, Köthe J, Wirths B, Sigge A, Poppert S. Superiority of molecular techniques for identification of gram-negative, oxidase-positive rods, including morphologically nontypical Pseudomonas aeruginosa, from patients with cystic fibrosis. J Clin Microbiol. 2005;43(8):4070–5. https://doi.org/10.1128/JCM.43.8.4070-4075.2005.
17. Dwivedi HP, Franklin S, Chandrasekaran S, Garner O, Traczewski MM, Beasley D, Pincus DH. Multicenter Clinical Evaluation of Vitek 2 Meropenem-Vaborbactam for Susceptibility Testing of Enterobacterales and Pseudomonas aeruginosa. J Clin Microbiol. 2022;60(1):e01610-21. https://doi.org/10.1128/jcm.01610-21.
18. Saud HM, Alaubydi MA. Production, extraction and partial purification of melanin pigment from pathogenic Klebsiella pneumoniae HM isolated from clinical samples. Int J Curr Microbiol Appl Sci. 2016;5(10):910–9. https://doi.org/10.20546/ijcmas.2016.510.098.
19. Pralea IE, Moldovan RC, Petrache AM, Ilieș M, Hegheș SC, Ielciu I, Nicoară R, Moldovan M, Ene M, Radu M. From extraction to advanced analytical methods: The challenges of melanin analysis. Int J Mol Sci. 2019;20(16):3943. https://doi.org/10.3390/ijms20163943.
20. Aghajanyan AE, Hambardzumyan AA, Minasyan EV, Tsaturyan AH, Paloyan AM, Avetisyan SV, Saghyan AS. Development of the technology for producing water-soluble melanin from waste of vinary production and the study of its physicochemical properties. Eur Food Res Technol. 2022;248(2):485–95. https://doi.org/10.1007/s00217-021-03843-6.
21. Bronze-Uhle ES, Batagin-Neto A, Xavier PH, Fernandes NI, De Azevedo ER, Graeff CF. Synthesis and characterization of melanin in DMSO. J Mol Struct. 2013;1047:102-108. https://doi.org/10.1016/j.molstruc.2013.04.058.
22. Zerrad A, Anissi J, Ghanam J, Sendide K, El Hassouni M. Antioxidant and antimicrobial activities of melanin produced by a Pseudomonas balearica strain. J Biotechnol Lett. 2014;5(2):87–94.
23. Singh S, Nimse SB, Mathew DE, Dhimmar A, Sahastrabudhe H, Gajjar A, Shinde PB. Microbial melanin: Recent advances in biosynthesis, extraction, characterization, and applications. Biotechnol Adv. 2021;53:107773. https://doi.org/10.1016/j.biotechadv.2021.107773.
24. Al Khatib M, Harir M, Costa J, Baratto MC, Schiavo I, Trabalzini L, Pollini S, Rossolini GM, Basosi R, Pogni R. Spectroscopic characterization of natural melanin from a Streptomyces cyaneofuscatus strain and comparison with melanin enzymatically synthesized by tyrosinase and laccase. Molecules. 2018;23(8):1916. https://doi.org/10.3390/molecules23081916.
25. Suthar M, Dufossé L, Singh SK. The enigmatic world of fungal melanin: a comprehensive review. J Fungi. 2023;9(9):891. https://doi.org/10.3390/jof9090891.
26. Roy S, Wu J, Cao J, Disu J, Bharadwaj S, Meinert-Spyker E, Wood S. Exploring the impact and influence of melanin on frequency-domain near-infrared spectroscopy measurements. J Biomed Opt. 2024;29(S3):S33310-S33310. https://doi.org/10.1117/1.JBO.29.S3.S33310.
27. Manivasagan P, Venkatesan J, Senthilkumar K, Sivakumar K, Kim SK. Isolation and characterization of biologically active melanin from Actinoalloteichus sp. MA-32. Int J Biol Macromol. 2013;58:263–74. https://doi.org/10.1016/j.ijbiomac.2013.04.041.
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