Two-Stage Fermentation for Torularhodin Production from Rhodotorula mucilaginosa Using Molasses
Main Article Content
Abstract
Torularhodin is a carotenoid of considerable interest due to its potent antioxidant activity and potential applications in the food, pharmaceutical, and cosmetic industries. In the study, the influence of culture conditions, including pH, temperature, and medium composition (molasses concentration and urea supplementation) on biomass formation and torularhodin biosynthesis in Rhodotorula mucilaginosa RL 231-1.51 was systematically investigated. Optimal biomass production was achieved at pH 6.5, 30 °C, and 10°Bx molasses, whereas maximum torularhodin accumulation was observed at pH 7.5, 25 °C, 22°Bx molasses, and 18 g/L urea. Implementation of a two-stage fermentation strategy, designed to decouple biomass growth from carotenoid production, resulted in a maximum torularhodin content of 2628.1 µg/g dry cell weight, corresponding to a yield of 91.98 mg/L. These findings demonstrate that the two-stage fermentation approach is an effective strategy to enhance torularhodin production and holds promise for industrial-scale applications.
Keywords
Molasses, Rhodotorula mucilaginosa, torularhodin.
Article Details
References
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[2] C. U. Mussagy, M. Gonzalez-Miquel, V. C. Santos-Ebinuma, and J. F. B. Pereira, Microbial torularhodin–a comprehensive review, Critical Reviews in Biotechnology, vol. 43, iss. 4, pp. 540–558, Apr. 2022. https://doi.org/10.1080/07388551.2022.2041540
[3] K. Varmira, A. Habibi, S. Moradi, and E. Bahramian, Statistical optimization of airlift photobioreactor for high concentration production of torularhodin pigment, Biocatalysis and Agricultural Biotechnology, vol. 8, pp. 197–203, Oct. 2016. https://doi.org/10.1016/j.bcab.2016.09.013
[4] A. El-Banna, Some factors affecting the production of carotenoids by Rhodotorula glutinis var. glutinis, Food and Nutrition Sciences, vol. 3, no. 1, pp. 64–71, Jan. 2012.
https://doi.org/10.4236/fns.2012.31011
[5] Y. T. Cheng and C. F. Yang, Using strain Rhodotorula mucilaginosa to produce carotenoids using food wastes, Journal of the Taiwan Institute of Chemical Engineers, vol. 61, pp. 270–275, Apr. 2016. https://doi.org/10.1016/j.jtice.2015.12.027
[6] A. Keskin, A. E. Ünlü, and S. Takaç, Utilization of olive mill wastewater for selective production of lipids and carotenoids by Rhodotorula glutinis, Applied Microbiology and Biotechnology, vol. 107, pp. 4973–4985, Jun. 2023. https://doi.org/10.1007/s00253-023-12625-x
[7] W. R. C. Machado, C. S. Murari, A. L. F. Duarte, and V. L. Del Bianchi, Optimization of agro-industrial coproducts (molasses and cassava wastewater) for the simultaneous production of lipids and carotenoids by Rhodotorula mucilaginosa, Biocatalysis and Agricultural Biotechnology, vol. 42, Mar. 2022, Art. no. 102342.
https://doi.org/10.1016/j.bcab.2022.102342
[8] M. Michelon, T. de Matos de Borba, R. da Silva Rafael, C. A. V. Burkert, and J. F. de Medeiros Burkert, Extraction of carotenoids from Phaffia rhodozyma: a comparison between different techniques of cell disruption, Food Science and Biotechnology, vol. 21, pp. 1–8, Feb. 2012. https://doi.org/10.1007/s10068-012-0001-9
[9] T. A. Pham, T. H. Luu, T. H. Dam, and K. A. To, Bioconversion of shrimp waste into functional lipid by a new oleaginous Sakaguchia sp., Molecular Biotechnology, vol. 67, pp. 3476–3484, Jan. 2024. https://doi.org/10.1007/s12033-023-01014-4
[10] Miller GL, Use of dinitrosalicylic acid reagent for determination of reducing sugar, Analytical Chemistry, vol. 3, no. 31, pp. 426–428, 1959. https://doi.org/10.1021/ac60147a030
[11] L. Zoz, J. C. Carvalho, V. T. Soccol, T. C. Casagrande, and L. Cardoso, Torularhodin and torulene: bioproduction, properties and prospective applications in food and cosmetics - a review, Brazilian Archives of Biology and Technology, vol. 58, no. 2, pp. 278–288, Apr. 2015. https://doi.org/10.1590/S1516-8913201400152
[12] N. Urano, A. Shirao, Y. Naito, M. Okai, M. Ishida, and M. Takashio, Molecular phylogeny and phenotypic characterization of yeasts with a broad range of pH tolerance isolated from natural aquatic environments, Advances in Microbiology, vol. 9, no. 1, pp. 56–73, Jan. 2019. https://doi.org/10.4236/aim.2019.91005
[13] L. D. Lario, O. S. Pillaca-Pullo, L. D. Sette, A. Converti, P. Casati, C. Spampinato, and A. Pessoa, Optimization of protease production and sequence analysis of the purified enzyme from the cold adapted yeast Rhodotorula mucilaginosa CBMAI 1528, Biotechnology Reports, vol. 28, Dec. 2020, Art. no. e00546. https://doi.org/10.1016/j.btre.2020.e00546
[14] K. D. Pham, Y. Shida, A. Miyata, T. Takamizama, Y. Suzuki, S. Ara, H. Yamazaki, K. Mori, S. Aburatani, H. Hirakawa, K. Tashiro, S. Kuhara, H. Takaku, and W. Ogasawara, Effect of light on carotenoid and lipid production in the oleaginous yeast Rhodosporidium toruloides, Bioscience, Biotechnology and Biochemistry, vol. 84, iss. 7, pp. 1501–1512, Jul. 2020. https://doi.org/10.1080/09168451.2020.1740581
[15] E. P. Cipolatti, R. D. Remedi, C. Santos Sá, A. B. Rodrigues, J. M. G. Ramos, C. A. V. Burkert, E. B. Furlong, and J. F. M. Burkert, Use of agroindustrial byproducts as substrate for production of carotenoids with antioxidant potential by wild yeasts, Biocatalysis and Agricultural Biotechnology, vol. 20, Jul. 2019, Art. no. 101208.
https://doi.org/10.1016/j.bcab.2019.101208
[16] N. Elfeky, M. Elmahmoudy, Y. Zhang, J. L. Guo, and Y. Bao, Lipid and carotenoid production by Rhodotorula glutinis with a combined cultivation mode of nitrogen, sulfur, and aluminium stress, Applied Science, vol. 9, iss. 12, Jun. 2019. https://doi.org/10.3390/app9122444
[17] R. Guo, T. Liu, C. Guo, G. Chen, J. Fan, and Q. Zhang, Carotenoid biosynthesis is associated with low-temperature adaptation in Rhodosporidium kratochvilovae, BMC Microbiology, vol. 22, Dec. 2022, Art. no. 319. https://doi.org/10.1186/s12866-022-02728-2
[18] R. Bao, N. Gao, Jing Lv, C. Ji, H. Liang, S. Li, C. Yu, Z. Wang, and X. Lin, Enhancement of torularhodin production in Rhodosporidium toruloides by Agrobacterium tumefaciens-mediated transformation and culture condition optimization, Journal of Agricultural and Food Chemistry, vol. 67, iss. 4, pp. 1156–1164, Jan. 2019.
https://doi.org/10.1021/acs.jafc.8b04667
[19] A. Ambrico, A new method for selective extraction of torularhodin from red yeast using CO2-SFE technique, Applied Biochemistry and Biotechnology, vol. 196, pp. 6473–6491, Feb. 2024. https://doi.org/10.1007/s12010-024-04884-9