Seyhan Ulusoy, Çağdaş Deniz Periz, Turgut Şekerler, Gülgün Tınaz

Ultrason Destekli Ekstraksiyon Yöntemi ile Elde Edilen Violasinin Antibakteriyel ve Antikanser Aktiviteleri

Viyolasin, özellikle Gram-negatif bakteri Chromobacterium violaceum tarafından üretilen ve antibakteriyel, antifungal, antioksidan ve antikanser etkilere sahip çeşitli bakteri türleri tarafından üretilen doğal mor bir pigmenttir. Bu çalışmada, viyolasin, Ultrason destekli (UA) ve mikrodalga destekli (MA) ekstraksiyon prosedürleri ile C. violaceum suşundan metanol, etanol, aseton ve etil asetat kullanılarak ekstrakte edilmiştir. Elde edilen viyolasin miktarı, yüksek performanslı sıvı kromatografisi (HPLC) kullanılarak belirlenmiştir. Daha sonra bu ekstraktların antibakteriyel ve antikanser aktiviteleri incelenmiştir. Bu çalışmada, viyolasin ilk kez UA ve MA ekstraksiyon yöntemleri kullanılarak elde edilmiştir. Viyolasin ekstrelerinin Staphylococcus aureus, S. aureus (MRSA) ve Bacillus cereus'a karşı antibakteriyel aktiviteye sahip olduğu belirlenmiştir. Viyolasin ekstresi rahim ağzı kanseri hücreleri (HeLa) üzerinde güçlü anti-kanser etkisi göstermiştir. Sonuçta, viyolasini ekstrakte etmek için, UA ve MA ekstraksiyon yöntemleri bakteri kültürlerine başarıyla uygulanmıştır.

Antibacterial and Anticancer Activities of Violacein Extracted Through Ultrasound-Assisted Extraction Method

Violacein is a natural violet pigment produced by various bacterial species, especially by the Gram-negative bacteriaChromobacterium violaceum. Violacein has antibacterial, antifungal, antioxidant and anticancer activities and hasvarious applications in food, cosmetic, textile and pharmaceutical industries. In this study, violacein was extractedfrom the Chromobacterium violaceum culture through ultrasonic-assisted (UA) and microwave-assisted (MA)extraction methods using methanol, ethanol, acetone, and ethyl acetate as solvent. Violacein was quantified usingHigh Performance Liquid Chromatography (HPLC). Then, antibacterial and anticancer activities of these extracts wereinvestigated. In this study, for the first time, the violacein extract was obtained by using UA and MA extractionmethods. The highest violacein concentration could be achieved by UAE method with methanol. It was found out thatviolacein extracts had an antibacterial effect against Staphylococcus aureus, S. aureus (MRSA) and Bacillus cereus.The violacein extract had a strong anticancer effect on cervical cancer (HeLa) cells. Finally, the UA and MA extractionmethods were successfully applied to bacterial cultures in order to extract the violacein.

PDF

___

[1] Choi, S.Y., Kim, S., Lyuck, S., Kim, S.B., Mitchell, R.J. (2015). High-level production of violacein by the newly isolated Duganella violaceinigra str. NI28 and its impact on Staphylococcus aureus. Scientific Reports, 5, 15598.
[2] Moss, M.O., Ryall, C. (1981). Distribution of chromobacteria in a lowland river. Microbial Ecology, 7(2), 139-149.
[3] Lichstein, H.C., Van De Sand, V.F. (1945). Violacein, an antibiotic pigment produced by Chromobacterium violaceum. The Journal of Infectious Diseases, 76(1), 47-51.
[4] Jude, B.A., Tanner, J., Koko, T., McLaughlin, E.C. (2012). Analysis, characterization, and synthesis of violacein from Janthinobacterium isolate extracts. In Abstracts of Papers of the American Chemical Society (Vol. 244). 1155 16TH ST, NW, Washıngton, DC 20036 USA: Amer. Chemical Soc.
[5] De Carvalho, D.D., Costa, F.T., Duran, N., Haun, M. (2006). Cytotoxic activity of violacein in human colon cancer cells. Toxicology in Vitro, 20(8), 1514- 1521.
[6] Durán, N., Justo, G.Z., Durán, M., Brocchi, M., Cordi, L., Tasic, G.R., Castro, L., Nakazato, G. (2016). Advances in Chromobacterium violaceum and properties of violacein-Its main secondary metabolite: A review. Biotechnology Advances, 34(5), 1030-1045.
[7] Durán, N., Menck, C.F. (2001). Chromobacterium violaceum: a review of pharmacological and industiral perspectives. Critical Reviews in Microbiology, 27(3), 201-222.
[8] Nakamura, Y., Asada, C., Sawada, T. (2003). Production of antibacterial violet pigment by psychrotropic bacterium RT102 strain. Biotechnology and Bioprocess Engineering, 8(1), 37-40.
[9] Bromberg, N., Durán, N. (2001). Violacein transformation by peroxidases and oxidases: implications on its biological properties. Journal of Molecular Catalysis B: Enzymatic, 11(4-6), 463- 467.
[10] Subramaniam, S., Ravi, V., Sivasubramanian, A. (2014). Synergistic antimicrobial profiling of violacein with commercial antibiotics against pathogenic micro-organisms. Pharmaceutical Biology, 52(1), 86-90.
[11] Mendes, A.S., de Carvalho, J.E., Duarte, M.C., Durán, N., Bruns, R.E. (2001). Factorial design and response surface optimization of crude violacein for Chromobacterium violaceum production. Biotechnology Letters, 23(23), 1963-1969.
[12] Durán, N., Justo, G.Z., Ferreira, C.V., Melo, P.S., Cordi, L., Martins, D. (2007). Violacein: properties and biological activities. Biotechnology and Applied Biochemistry, 48(3), 127-133.
[13] Durán, M., Ponezi, A.N., Faljoni-Alario, A., Teixeira, M.F., Justo, G.Z., Durán, N. (2012). Potential applications of violacein: a microbial pigment. Medicinal Chemistry Research, 21(7): 1524-1532.
[14] Venil, C.K., Aruldass, C.A., Abd Halim, M.H., Khasim, A.R., Zakaria, Z.A., Ahmad, W.A. (2015). Spray drying of violet pigment from Chromobacterium violaceum UTM 5 and its application in food model systems. International Biodeterioration & Biodegradation, 102, 324-329.
[15] Nakamura, Y., Sawada, T., Morita, Y., Tamiya, E. (2002). Isolation of a psychrotrophic bacterium from the organic residue of a water tank keeping rainbow trout and antibacterial effect of violet pigment produced from the strain. Biochemical Engineering Journal, 12(1), 79-86.
[16] Zhang, Q.W., Lin, L.G., Ye, W.C. (2018). Techniques for extraction and isolation of natural products: a comprehensive review. Chinese Medicine, 13(1), 20.
[17] Kaufmann, B., Christen, P. (2002). Recent extraction techniques for natural products: microwave‐assisted extraction and pressurised solvent extraction. Phytochemical Analysis: An International Journal of Plant Chemical and Biochemical Techniques, 13(2), 105-113.
[18] Wang, L., Weller, C.L. (2006). Recent advances in extraction of nutraceuticals from plants. Trends in Food Science & Technology, 17(6), 300-312.
[19] Blosser, R.S., Gray, K.M. (2000). Extraction of violacein from Chromobacterium violaceum provides a new quantitative bioassay for N-acyl homoserine lactone autoinducers. Journal of Microbiological Methods, 40(1), 47-55.
[20] Holder, I.A., Boyce, S.T. (1994). Agar well diffusion assay testing of bacterial susceptibility to various antimicrobials in concentrations non-toxic for human cells in culture. Burns, 20(5), 426-429.
[21] Batista, A.H., Moreira, A.C., De Carvalho, R.M., Sales, G.W., Nogueira, P.C., Grangeiro, T.B., Medeiros S.C., Silveira E.V., Nogueira, N.A. (2017). Antimicrobial effects of violacein against planktonic cells and biofilms of Staphylococcus aureus. Molecules, 22(10), 1534.
[22] Cazoto, L.L., Martins, D., Ribeiro, M.G., Durán, N., Nakazato, G. (2011). Antibacterial activity of violacein against Staphylococcus aureus isolated from bovine mastitis. The Journal of Antibiotics, 64(5), 395-397.
[23] Aranda, S., Montes-Borrego, M., Landa, B.B. (2011). Purple-pigmented violacein-producing Duganella spp. inhabit the rhizosphere of wild and cultivated olives in southern Spain. Microbial Ecology, 62(2), 446-459.
[24] Im, H., Choi, S.Y., Son, S., Mitchell, R.J. (2017). Combined application of bacterial predation and violacein to kill polymicrobial pathogenic communities. Scientific Reports, 7(1), 1-10.
[25] Choi, S.Y., Lim, S., Cho, G., Kwon, J., Mun, W., Im, H., Mitchell, R.J. (2020). Chromobacterium violaceum delivers violacein, a hydrophobic antibiotic, to other microbes in membrane vesicles. Environmental Microbiology, 22(2), 705-713.
[26] Saraiva, V.S., Marshall, J.C., Cools-Lartigue, J., Burnier Jr, M.N. (2004). Cytotoxic effects of violacein in human uveal melanoma cell lines. Melanoma Research, 14(5), 421-424.
[27] Bromberg, N., Dreyfuss, J.L., Regatieri, C.V., Palladino, M.V., Duran, N., Nader, H.B., Justo, G.Z. (2010). Growth inhibition and pro-apoptotic activity of violacein in Ehrlich ascites tumor. Chemicobiological Interactions, 186(1), 43-52.