Document Type : Original Paper

Authors

1 Ph.D. Student, Department of biology, Faculty of Basic Science, Science and Research Branch, Islamic Azad University, Tehran, Iran

2 Department of Microbiology, Faculty of Basic Siences, North Tehran Branch, Islamic Azad University, Tehran, Iran

3 Department of Microbiology, Faculty of Basic Science, Saveh Branch, Islamic Azad University, Markazi, Iran

Abstract

Biological sources of pigments receive major attention nowadays because of the stringent rules and regulations applied to chemically synthesized pigments. The aims of this study were isolating carotenoids producing Micrococcus spp. and Rhodotorula spp. from soil sources, optimizing the culture conditions for biomass and carotenoids production, molecular cloning of Crt gene, characterizing yielded pigment, and investigating their anti-cancer activities on human breast cancer cells. Carotenoid producing strains, M. luteus and R. mucilaginosa, were isolated from the soil and sediment samples in Kerman Province, Iran; they were identified using 16srDNA analysis. Optimum conditions for biomass and carotenoids production were determined. Further, Crt gene cloned into pTG19 vector and the effect of carotenoid pigments on MCF-7 breast cancer cell line were evaluated. The optimum growth and carotenoids production by strains were 25°C and pH 7.0 for M. luteus, and 25°C and pH 6.5 for R. mucilaginosa. FTIR and TLC analysis showed high similarity of extracted pigments with carotenoids. The expression levels of Crt genes mRNA found to be approximately 5-fold higher in transformed E. coli in comparison with M. luteus and R. mucilaginosa (p <0.001). MCF-7 cells viability decreased in a concentration and time dependent manner (p <0.05). The doses IC50 against MCF-7 cells for yellow and pink pigments were calculated to be 1426.69 µg/ml (r2=0.95) and 1412.1 µg/ml (r2=0.92), respectively. Microorganisms presented in this study can be used as potential sources of commercial carotenoids production and antitumor metabolites.

Keywords

1.   Netzer R, Stafsnes MH, Andreassen T, Goksøyr A, Bruheim P, Brautaset T. Biosynthetic pathway for γ-cyclic sarcinaxanthin in Micrococcus luteus: Heterologous expression and evidence for diverse and multiple catalytic functions of C50 carotenoid cyclases. J Bacteriol. 2010; 192(21):5688-99

2.   Stafsnes MH. Characterization and exploitation of a marine microbial culture collection:–a special focus on carotenoid producing heterotrophic bacteria. Doctoral thesis., 2013. http://hdl.handle.net/11250/245930

3.   Goodwin TW. Chemistry and biochemistry of plant pigments: Academic Press; 1976.

4.   Song MJ, Bae J, Lee DS, Kim CH, Kim JS, Kim SW, et al. Purification and characterization of prodigiosin produced by integrated bioreactor from Serratia sp. KH-95. J Biosci Bioengin. 2006;101(2):157-61.

5.   Barnett JA, Payne RW, Yarrow D. Yeasts: characteristics and identification: Cambridge University Press; 1983.

6.   Zhao Y, Guo L, Xia Y, Zhuang X, Chu W. Isolation, Identification of Carotenoid-Producing Rhodotorula sp. from Marine Environment and Optimization for Carotenoid Production. Mar Drugs. 2019;17(3):161.

7.   Muthezhilan R, Ragul R, Pushpam R, Narayanan RL, Hussain AJ. Isolation, optimization and extraction of microbial pigments from marine yeast Rhodotorula Sp (Amby109) as food colourants. Biosci Biotechnol Res Asia. 2014;11:271-8.

8.   Surekha P, Dhanya P, Sarath M, Pradeep S, Benjamin S. Micrococcus luteus strain BAA2, a novel isolate produces carotenoid pigment. Electron J Biol. 2016;12(1):83-9.

9.   Al-Wandawi H. Carotenoid Biosynthesis in Micrcoccus luteus Grown in the Presence of Different Concentrations of Nicotine. Int J Pure Appl Sci Technol. 2014;24(1).

10. Elsanhoty R, Al-Turki AI, Abdel-Razik MM. Production of carotenoids from rhodotorula mucilaginosa and their applications as colorant agent in sweet candy. J Food Agriculture Environ. 2017;15:21-6.

 

11. Taskin M, Sisman T, Erdal S, Kurbanoglu EB. Use of waste chicken feathers as peptone for production of carotenoids in submerged culture of Rhodotorula glutinis MT-5. Eur Food Res Technol. 2011;233(4):657.

12. Malisorn C., Suntornsuk W. Optimization of β-carotene production by Rhodotorula glutinis DM28 in fermented radish brine. Bioresour Technol. 2008;99(7):2281-7.

13. Akinnibosun F, Omorodion S. Isolation and Characterization of Antibiotic-Resistant Bacteria from Pesticide-Contaminated Agricultural Soils in Edo State, Nigeria. NISEB J. 2017;16(1).

14. Abbes M, Baati H, Guermazi S, Messina C, Santulli A, Gharsallah N, et al. Biological properties of carotenoids extracted from Halobacterium halobium isolated from a Tunisian solar saltern. BMC Complement Altern Med. 2013;13(1):255.

15. Shree GS, Prasad KY, Arpitha H, Deepika U, Kumar KN, Mondal P, et al. β-carotene at physiologically attainable concentration induces apoptosis and down-regulates cell survival and antioxidant markers in human breast cancer (MCF-7) cells. Mol Cell Biochem. 2017;436(1-2):1-12.

16. Vijay K, Sowmya PRR, Arathi BP, Shilpa S, Shwetha HJ, Raju M, et al. Low-dose doxorubicin with carotenoids selectively alters redox status and upregulates oxidative stress-mediated apoptosis in breast cancer cells. Food Chem Toxicol. 2018;18:675-90.