Cytotoxic effects and Identification of bioactive metabolites from crude extract of potential probiotic bacterium, Bacillus amyloliquefacians CS4, isolated from the gut of freshwater fish Channa striata (Bloch, 1793)

Nobel Surya Pandi Durai R¹ , Aiswarya D² , Amutha V³ , Sethuraman G⁴ , erumal P⁵

Section:Research Paper, Product Type: Isroset-Journal
Vol.6 , Issue.3 , pp.14-19, Jun-2019

CrossRef-DOI:   https://doi.org/10.26438/ijsrbs/v6i3.1419

Online published on Jun 30, 2019

Copyright © Nobel Surya Pandi Durai R, Aiswarya D, Amutha V, Sethuraman G, Perumal P . This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
 

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Abstract :
In the present study, the crude secondary metabolites were extracted from the potential probiotic bacterium, Bacillus amyloliquefacians CS4 using solvent ethyl acetate and the cytotoxicity of bacterial extract was assessed against HeLa cell line through MTT assay. The crude extract exhibited cytotoxicity effect on HeLa cell lines with the effective half maximal inhibitory concentration (IC50) value of 121.65µg/ml. The brine shrimp toxicity assay of crude extract revealed half maximal lethal concentration (LC50) value at 93.377μg/ml against Artemia salina. The zebra fish (Danio rerio) embryos were treated with different concentrations of extract upto 72 hpf and was found lowest survivability (36.6 %) at 200μg/ml concentration. The bioactive compounds and their functional groups of crude extract were identified using GC-MS and FTIR analyses. The secondary metabolites produced by B. amyloliquefacians CS4 have caused potential cytotoxic effects on cancer cell line and these compounds could be possibly used for the development of therapeutic agents, in relation to cancer drug discovery.

Key-Words / Index Term :
Anticancer activity, Bacillus amyloliquefacians, Secondary metabolites, Brine shrimp assay, GC-MS

References :
[1]. M.S. Butler, “Natural products to drugs: Natural productderived compounds in clinical trials”, Natural Product Report, Vol. 25, pp-475–516, 2008.
[2]. W.F. Safari, E. Chasanah, A.T. Wahyudi, “Antibacterial and anticancer activities of marine bacterial extracts and detection of genes for bioactive compounds synthesis”, International Journal of Pharmacy and Pharmaceutical Science, Vol. 8, Issue. 2, pp. 55-59, 2016.
[3]. H. Sugita, Y. Hirose, N. Matsuo, and Y. Deguchi, “Production of the antibacterial substance by Bacillus sp. Strain NM 12, an intestinal bacterium of Japanese coastal fish”, Aquaculture. Vol. 165, pp. 269–280, 1998.
[4]. R. Somayeh, R. Ali, H. Mehrdad and N. Tahereh, “Artemia salina as a model organism in toxicity assessment of nanoparticles”. DARU Journal of Pharmaceutical Sciences, Vol. 23, DOI 10.1186/s40199-015-0105-x, 2015
[5]. A. K. Neu, M. Månsson, L. Gram, M.J. Prol-García, “Toxicity of Bioactive and Probiotic Marine Bacteria and Their Secondary Metabolites in Artemia sp. and Caenorhabditis elegans as Eukaryotic Model Organisms”, Applied and Environmental Microbiology, Vo.1, pp. 146–153, 2013.
[6]. D.J. Newman, and G.M. Cragg, “Natural Products as Sources of New Drugs over the Last 25 Years”, Journal of Natural Products, Vol. 70, pp. 461-477, 2007.
[7]. X.H. Chen, A. Koumoutsi, R. Scholz, A. Eisenreich, K. Schneider, I. Heinemeyer, B. Morgenstern, B. Voss, W.R. Hess, O. Reva, H. Junge, B. Voigt, P.R. Jungblut, J. Vater, R. Sussmuth, H. Liesegang, A. Strittmatter, G. Gottschalk and R. Borriss, “Comparative analysis of the complete genome sequence of the plant growth-promoting bacterium Bacillus Amyloliquefaciens Fzb42”, Nature Biotechnology, Vol. 25, pp. 1007–1014, 2007.
[8]. R.K. Ganguly, S. Midya, and S.K. Chakraborty, “Antioxidant and Anticancer Roles of a Novel Strain of Bacillus anthracis Isolated from Vermicompost Prepared from Paper Mill Sludge”, BioMed Research International. Vol. 1, article ID 1073687, pp. 7, 2018.
[9]. E. Hinarejos, M. Castellano, I. Rodrigo, Bacillus subtilis IAB/BS03 as a potential biological control agent, European Journal of Plant Pathology, Vol. 146, pp. 597–608, 2016.
[10]. R. Nagel, “Dar T: the embryo test with the zebrafish Danio rerioa general model in ecotoxicology and toxicology”. Altex, Vol. 19, pp. 38–48, 2002.
[11]. E. Sansinenea and A. Ortiz, “Secondary metabolites of soil Bacillus spp.”, Biotechnology Letters, Vol. 33, pp. 1523–1538, 2011.
[12]. G.H. Park, H.M. Song, Y.S. Kim, Y. Jeon, J.S. Koo, H.J. Jeong, & J.B. Jeong, “Anti-cancer activity of Bacillus amyloliquefaciens AK-0 through cyclin D1 proteasomal degradation via GSK3 dependent phosphorylation of threonine-286”, Die Pharmazie, Vol. 72, pp. 348–354, 2017.
[13]. R. Ramasubburayan, S. Sumathi, D. MagiBercy, G. Immanuel, and A. Palavesam, “Antimicrobial, antioxidant and anticancer activities of mangrove associated bacterium Bacillus subtilis RG”, Biocatalysis and Agricultural Biotechnology, Vol. 4, pp. 158–165, 2015.
[14]. D. R. Ammons, J.D. Short, J. Bailey, G. Hinojosa, L. Tavarez, M. Salazar and J.N. Rampersad, “Anti-cancer Parasporin Toxins are Associated with Different Environments: Discovery of Two Novel Parasporin 5-like Genes”, Current Microbiology, 2016 https://doi.org/10.1007/s00284-015-0934-3.
[15]. A. Manilal, S. Sujith, G.S. Kiran, J. Selvin, and C. “Shakir, Cytotoxic potentials of red algae, Laurencia brandenii collected from the Indian coast”, Global Journal of Pharmacology, Vol. 3, pp. 90–94, 2009.
[16]. M.O. Ullah, M. Haque, K.F. Urmi, A.H.M. Zulfiker, E.S. Anita, M. Begum, K. Hamid, “Anti–bacterial activity and brine shrimp lethality bioassay of methanolic extracts of fourteen different edible vegetables from Bangladesh. Asian Pacific Journal of Tropical Biomedicine, Vol. 3, pp. 1–7, 2013.
[17]. R. Embry, S. E. Belanger, T. A. Braunbeck, “The fish embryo toxicity test as an animal alternative method in hazard and risk assessment and scientific research”, Aquatic Toxicology, Vol. 97, pp. 79–87, 2010.
[18]. S. Glaberman, S. Padilla, and M.G. Barron, “Evaluating the zebrafish embryo toxicity test for pesticide hazard screening,” Environmental Toxicology and Chemistry”, Vol. 36, pp. 1221–1226, 2017.
[19]. G. Balasubramani, D. Arul, C. Kamaraj, P. Deepak, D. Aiswarya, K. Jothimani, P. Santhanam, P. Perumal, “Comparative extraction of Salmonella bongori derived metabolites and their toxicity on bacterial pathogens, mosquito-larvae, zebrafish-embryo and brine-shrimp: A modified approach”, Ecotoxicology and Environmental Safety, Vol. 169, pp. 192–206, 2019.
[20]. M. Gopi, N. Balachandran Dhayanithi, K. Nanthini Devi, T. T. Ajith Kumar, “Marine natural product, Pyrrolo[1,2-a]pyrazine-1,4-dione, hexahydro-(C7H10N2O2) of antioxidant properties from Bacillus species at Lakshadweep archipelago”, Journal of Coastal Life Medicine, Vol. 2, pp. 636-641, 2014.