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Nikanova D.A., Artemyeva O.A., Kolodina E.N., Dovydenkova M.V., Berezova K.A., Mikhel I.M., Barashkova A.S., Rogozhin E.A. Saprophytic yeast Rhodotorula mucilaginosa: molecular identification, morphobiochemical properties and carotenoid production. Sel'skokhozyaistvennaya Biologiya [Agricultural Biology], 2025, Vol. 60, № 6, p. 1126-1141.
(how to cite this article)

doi: 10.15389/agrobiology.2025.6.1126eng

UDC: 579.64:663.12

Acknowledgements:
Supported financially by the Russian Science Foundation, grant No. 23-16-00167

 

SAPROPHYTIC YEAST Rhodotorula mucilaginosa: MOLECULAR IDENTIFICATION, MORPHOBIOCHEMICAL PROPERTIES AND CAROTENOID PRODUCTION

D.A. Nikanova1 ✉, O.A. Artemyeva1, E.N. Kolodina1,
M.V. Dovydenkova1, K.A. Berezova1, I.M. Mikhel2, 3,
A.S. Barashkova3, 4, E.A. Rogozhin3, 4

1Ernst Federal Research Center for Animal Husbandry, 60, pos. Dubrovitsy, Podolsk District, Moscow Province, 142132 Russia, e-mail dap2189@gmail.com (✉ corresponding author), vijmikrob@mail.ru, kolodin77@mail.ru, majra_2005@list.ru, voshaisreal@yandex.ru;
2All-Russian Research Institute of Agricultural Biotechnology, 42, ul. Timiryazevskaya, Moscow, 127550 Russia, e-mail joseph.mikhel@yandex.ru;
3Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry RAS, 16/10, ul. Miklukho-Maklaya, Moscow, 117997 Russia, e-mail barashkova.an@gmail.com, rea21@list.ru;
4All-Russian Research Institute of Plant Protection, 3, sh. Podbel’skogo, St. Petersburg, 196608 Russia, e-mail barashkova.an@gmail.com, rea21@list.ru

ORCID:
Nikanova D.A. orcid.org/0000-0001-5164-244X
Berezova K.A. orcid.org/0009-0005-9556-1947
Artemyeva O.A. orcid.org/0000-0001-7706-4182
Mikhel I.M. orcid.org/0000-0002-7930-8862
Kolodina E.N. orcid.org/0000-0002-4017-3390
Barashkova A.S. orcid.org/0000-0002-3455-2335
Dovydenkova M.V. orcid.org/0000-0002-3093-4117
Rogozhin E.A. orcid.org/0000-0003-0659-9547

Final revision received July 31, 2025
Accepted September 29, 2025

Rhodotorula is a genus of saprophytic yeasts of the family Sporidiobolaceae, widespread in air, soil, lakes, seawater, milk, food products, and capable of colonizing protozoa, plants, humans, and other mammals. Three species of Rhodotorula (R. mucilaginosa, R. glutinis, and R. minuta) are considered opportunistic pathogens and are the most common causative agents of infections in immunocompromised individuals. However, in recent years, Rhodotorula mucilaginosa yeasts have increasingly been used in biotechnology for carotenoid production and in feeding agricultural animals. Carotenoid pigments act as antioxidants and possess anticancer properties, including immunomodulatory and oncoprotective effects, which contribute to the normalization of reproductive function, growth, and development in farm animals and poultry. This study provides the first comprehensive investigation of R. mucilaginosa yeasts isolated from the gastrointestinal tract of farm animals, including species identification based on the 5.8S-ITS rDNA fragment, assessment of intraspecific morphobiochemical variability, determination of optimal growth temperature, and identification of the most productive carotenoid-producing strains on a non-enriched medium. The aim of the study was to isolate pigmented yeasts from the gastrointestinal contents of farm animals and poultry, followed by species identification, cultivation, biomass production, assessment of the total carotenoid content and their concentration in biomass, as well as the identification of the most promising isolates for further use as components of a feed additive. Experiments were conducted from 2023 to 2025. Yeast isolates were obtained from the gastrointestinal tract contents of Black-and-White cattle, crossbred F2 pigs (Large White × Landrace) × Duroc, and broiler chickens of the Smena 9 cross (Ernst Federal Research Center for Animal Husbandry). Isolation was performed on Sabouraud dextrose agar (HiMedia Laboratories Pvt. Ltd., India). For microscopic analysis, standard methylene blue staining was used to determine cell morphology. Morphological characteristics and culture purity were assessed using a Nicon ECLIPSE Ci-L light microscope (Nikon, Japan). The biochemical profile was determined using the KB009R HiCarbo test system (HiMedia Laboratories Pvt. Ltd., India) and specific sugars. Polymerase chain reaction (PCR) was performed using a Mini Amp Plus thermal cycler (Thermo Fisher Scientific, Singapore). For amplification of the 5.8S-ITS rDNA fragment and determination of its primary nucleotide sequence, the ITS1 (5´-TCC GTA GGT GAA CCT GCG G-3´) and ITS4 (5´-TCC TCC GCT TAT TGA TAT GC-3´) primers were used. Pigments were extracted from samples containing frozen raw yeast biomass (5-20 g) from each isolate. Carotenoid content in the extracts was quantified spectrophotometrically using a UV-1800 spectrophotometer (Shimadzu Corp., Japan). Absorption spectra were recorded in the 300-600 nm range. All isolates utilized glucose, dextrose, mannose, raffinose, sucrose, and fructose as carbon sources for fermentation. Colonies of all isolates grown on DAP medium exhibited an orange-red pigmentation, were round, convex, with smooth surfaces, even edges, and homogeneous structure. Optimal growth temperature ranged from 19 to 30 °C. The largest cell areas were observed in strains Chli 32 (4.364 µm²) and Sr 16 (4.714 mm2), while the highest elongation coefficient (Ec) was found in strains Pf 22 (1.821 at 3.069±0.100 mm cell length and 1.685±0.033 mm width) and Chc 8 (1.685 at 2.727±0.098 mm length and 1.619±0.034 mm width). Strains Chc 65 and Pf 13 had cell areas of 4.121 and 4.298 mm2, respectively, with Ec values of 1.414 and 1.520, indicating a more elongated cell shape. The smallest cell area was recorded in strain Chc 6 (1.786 mm2). Carotenoids were successfully extracted from the biomass of all isolates and analyzed spectrophotometrically. Based on the results, six isolates (Chli 32, Chli 49, Chc 65, Cm 17, Cr 32, Sr 40) were identified as having the highest carotenoid content (22.86; 21.71; 18.05; 15.08; 11.74; 11.24 mg/g of raw biomass). These isolates seem to be the most promising for further optimization of cultivation parameters and enhancement of productivity.

Keywords: Rhodotorula mucilaginosa, extraction, organic solvents, carotenoids, spectrophotometric analysis.

 

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