Polypeptide Antibiotic Produced by a Thermo-Halophilic Bacterium from Pria Laot Sabang 76 Isolate
DOI:
https://doi.org/10.25077/jrk.v15i1.634Keywords:
Thermo-halophilic bacteria, polypeptide antibiotic, Pria Laot Sabang, PolymyxinAbstract
Extremophiles are known to produce antibiotics. This study aimed to isolate and purify polypeptide antibiotics from Pria Laot Sabang (PLS) 76, a bacterium isolated from an undersea fumarole. The antibiotic production was done by cultivating PLS 76 in a 2.5% TSB liquid medium and incubated at 70°C, 150 rpm for 112 hours. Ninhydrin assay on the crude supernatant after centrifugation indicated that the PLS 76 produced a polypeptide antibiotic. The supernatant was partitioned using methanol. The antibiotic activity was tested using the Kirby-Bauer method, and the inhibition zone was about 12 mm against Escherichia coli and Staphylococcus aureus. The methanol fraction was purified further by Preparative Thin Layer Chromatography and produced several fractions. Fractions with the same Rf were pooled and subjected to the ninhydrin assay. Fraction A, which gave a positive result after the ninhydrin assay, showed an inhibition zone of only 7 mm against E. coli and S. aureus. The Quadrupole Time-of-Flight (QTOF) Mass Spectrometry data indicated that the antibiotic's structure was similar to Polymyxin B2, an antibiotic of a polypeptide group.
References
Nikaido, H., Multidrug Resistance in Bacteria. Annu. Rev. Biochem., 78(1): 119–146 (2009).
Kaur, H., Onsare, J. G., Sharma, V. & Arora, D. S., Isolation, purification and characterization of novel antimicrobial compound 7-methoxy-2,2-dimethyl-4-octa-4′,6′-dienyl-2H-napthalene-1-one from Penicillium sp. and its cytotoxicity studies. AMB Express, 5(1): 40 (2015).
Lange, L. & Lopez, C. S., Protection agents from nature: Natural products and analogues. Cambridge, UK, Royal Society of Chemistry, (1996).
Jiao, K., Gao, J., Zhou, T., Yu, J., Song, H., Wei, Y. & Gao, X., Isolation and purification of a novel antimicrobial peptide from Porphyra yezoensis. J. Food Biochem., 43(7): (2019).
Cheng, A.-C., Lin, H.-L., Shiu, Y.-L., Tyan, Y.-C. & Liu, C.-H., Isolation and characterization of antimicrobial peptides derived from Bacillus subtilis E20-fermented soybean meal and its use for preventing Vibrio infection in shrimp aquaculture. Fish Shellfish Immunol., 67: 270–279 (2017).
Lei, J., Sun, L., Huang, S., Zhu, C., Li, P., He, J., Mackey, V., et al., The antimicrobial peptides and their potential clinical applications. Am. J. Transl. Res., 11(7): 3919–3931 (2019).
Marshall, S. H. & Arenas, G., Antimicrobial peptides: A natural alternative to chemical antibiotics and a potential for applied biotechnology . Electronic Journal of Biotechnology , 6: 271–284 (2003).
Akhmaloka, A., Suharto, A., Nurbaiti, S., Tika, I. N. & Warganegara, F. M., Ribotyping Identification of Thermophilic Bacterium from Papandayan Crater. J. Eng. Technol. Sci., 38(1): 1–10 (2006).
Grégoire, P., Bohli, M., Cayol, J.-L., Joseph, M., Guasco, S., Dubourg, K., Cambar, J., et al., Caldilinea tarbellica sp. nov., a filamentous, thermophilic, anaerobic bacterium isolated from a deep hot aquifer in the Aquitaine Basin. Int. J. Syst. Evol. Microbiol., 61(6): 1436–1441 (2011).
Iqbalsyah, T. M., Nisak, C., Sartika, F., Saidi, N., & Febriani, F. Identification of Antibiotic in Ethyl Acetate Fraction Produced by A Local Isolate PLS 80 Isolated from Shallow Sea Fumaroles. IOP Conference Series: Earth and Environmental Science, 276(1), 012006. (2019).
Başbülbül Özdemir, G. & Biyik, H. H., Isolation and Characterization of a Bacteriocin-Like Substance Produced by Geobacillus toebii Strain HBB-247. Indian J. Microbiol., 52(1): 104–108 (2012).
Ebrahimipour, G. hossein., Khosravibabadi, Z., Sadeghi, H. & Aliahmadi, A., Isolation, Partial Purification and Characterization of an Antimicrobial Compound, Produced by Bacillus atrophaeus. Jundishapur J. Microbiol., 7(8): (2014).
Sánchez, L. A., Sierra, M. G., Siñeriz, F. & Delgado, O., Andrimid production at low temperature by a psychrotolerant Serratia proteamaculans strain. World J. Microbiol. Biotechnol., 29(10): 1773–1781 (2013).
Sawale, A. A., Kadam, T. & Mitkare, S., Isolation and characterization of secondary metabolites from halophilic bacillus species from marin drive in mumbai. J. Appl. Pharm. Sci., 3: 182–188 (2013).
Hussain, M. Y., Ali-Nizam, A. A., & Abou-Isba, S. M. Antibacterial Activities (Bacitracin A and Polymyxin B) of Lyophilized Extracts from Indigenous Bacillus subtilis Against Staphylococcus aureus. 10 (3) (2017).
Atlas, R. M., Handbook of Microbiological Media. CRC Press, (2010). doi:10.1201/EBK1439804063
Hancu, G., Simon, B., Kelemen, H., Rusu, A., Mircia, E. & Gyeresi, A., Thin layer chromatographic analysis of Beta-lactam antibiotics. Adv. Pharm. Bull., 3(2): 367–71 (2013).
Iqbalsyah, T.M., Saidi, N., Maulyna, N., Helwati, H., Aura N., F., Skrining Antibiotik Dari Bakteri Thermo-Halofilik Isolat Pria Laot Sabang (PLS A dan 76). in Prosiding seminar nasional xx perhimpunan biokimia dan biologi molekuler Indonesia (pbbmi) “Current Excitement in Biochemistry and Molecular Biology for Agriculture and Medicine, Fakultas Kedokteran Universitas Lampung, (2016).
Vijayakumari, S. J., Sasidharannair, N. K., Nambisan, B. & Mohandas, C., Optimization of media and temperature for enhanced antimicrobial production by bacteria associated with Rhabditis sp. Iran. J. Microbiol., 5(2): 136–41 (2013).
Kumar, R., Shrivastav, A. K., Singha, A. K., Kumar, P. & Nirmala, A., Antibiotic production from marine Strptomycyes sp. Int J Pharma Bio Sci, 3(4): 331–342 (2012).
Darmayasa, I, B, C., Daya Hambat Fraksinasi Ekstrak Sembung Delan (Sphaerantus Indicus L) Terhadap Bakteri Eschericia coli Dan Staphylococcus aureus. J. Biol. Udayana; Vol 12 No 2 J. Biol., 12(2): 74–77 (2008).
Pilonieta M. C., Erickson K. D., Ernst R. K., Detweiler C. S. (2009). A protein important for antimicrobial peptide resistance, YdeI/OmdA, is in the periplasm and interacts with OmpD/NmpC. J. Bacteriol. 191 7243–7252 (2009).
Waluyo. J.. Sugiharto. B.. dan Zaini. N. C.. Purifikasi dan Karakterisasi Protein Antibakteri dari Pheretima javanica. Jurnal Ilmu Dasar 8 (1) : 37-44 (2007).
Doran-Peterson, J. & Decrescenzo-Henriksen, E., Methods For Producing And Using Polymyxins. (2011).
Thomas, T. A., Broun, E. C., Abildskov, K. M., Kubin, C. J., Horan, J., Yin, M. T. & Cremers, S., High Performance Liquid Chromatography–Mass Spectrometry Assay for Polymyxin B1 and B2 in Human Plasma. Ther. Drug Monit., 34(4): 398–405 (2012).
Meng, M., Wang, L., Liu, S., Jaber, O. M., Gao, L., Chevrette, L. & Reuschel, S., Simultaneous quantitation of polymyxin B1, polymyxin B2 and polymyxin B1-1 in human plasma and treated human urine using solid phase extraction and liquid chromatography–tandem mass spectrometry. J. Chromatogr. B, 1012–1013: 23–36 (2016).
Ayoub Moubareck, C.Polymyxins and Bacterial Membranes: A Review of Antibacterial Activity and Mechanisms of Resistance. Membranes, 10 (8), 181 (2020)
Yu, Z., Qin, W., Lin, J., Fang, S. & Qiu, J., Antibacterial Mechanisms of Polymyxin and Bacterial Resistance. Biomed Res. Int., 2015: 1–11 (2015).
Downloads
Published
How to Cite
Issue
Section
Citation Check
License
This work is licensed under a Creative Commons Attribution 4.0 International License.
Please find the rights and licenses in Jurnal Riset Kimia (J. Ris. Kim). By submitting the article/manuscript of the article, the author(s) agree with this policy. No specific document sign-off is required.
1. License
The use the article will be governed by the Creative Commons Attribution license as currently displayed on Creative Commons Attribution 4.0 International License.Â
2. Author(s)' Warranties
The author warrants that the article is original, written by stated author(s), has not been published before, contains no unlawful statements, does not infringe the rights of others, is subject to copyright that is vested exclusively in the author and free of any third party rights, and that any necessary written permissions to quote from other sources have been obtained by the author(s).
3. User Rights
Under the Creative Commons license, the journal permits users to copy, distribute, and display the material for any purpose. Users will also need to attribute authors and J. Ris. Kim on distributing works in the journal and other media of publications.
4. Rights of Authors
Authors retain all their rights to the published works, such as (but not limited to) the following rights;
- Copyright and other proprietary rights relating to the article, such as patent rights,
- The right to use the substance of the article in own future works, including lectures and books,
- The right to reproduce the article for own purposes,
- The right to self-archive the article,
- The right to enter into separate, additional contractual arrangements for the non-exclusive distribution of the article's published version (e.g., post it to an institutional repository or publish it in a book), with an acknowledgment of its initial publication in this journal.
5. Co-Authorship
If the article was jointly prepared by more than one author, any authors submitting the manuscript warrants that he/she has been authorized by all co-authors to be agreed on this copyright and license notice (agreement) on their behalf, and agrees to inform his/her co-authors of the terms of this policy. J. Ris. Kim will not be held liable for anything that may arise due to the author(s) internal dispute. J. Ris. Kim will only communicate with the corresponding author.