Exploring the lipids, carotenoids, and vitamins content of Rhodotorula glutinis with selenium supplementation under lipid accumulating and growth proliferation conditions.

BACKGROUND: Rhodotorula glutinis, a specific type of yeast, has been recognised as a superior resource for generating selenium-enriched biomass that possesses exceptional nutritional and functional attributes. The purpose of this investigation was to assess the effect of sodium selenite at different concentrations on lipid and carotenoid synthesis, as well as the growth of R. glutinis. METHODS: The lipid's fatty acid composition was determined using gas chromatography (GC). The vitamins were detected by high-performance liquid chromatography (HPLC). Transmission electron microscopy was used to detect the structural modification of yeast cells caused by the addition of sodium selenite to the growth medium, as well as the accumulation of elemental selenium in the yeast cells. RESULTS: The yeast cells demonstrated the ability to endure high concentrations of sodium selenite under lipid accumulation (LAM) and growth-promoting (YPD) conditions. 25.0 mM and 30.0 mM, respectively, were published as the IC50 values for the LAM and YPD conditions. In both growth media, 1 mM sodium selenite boosted lipid synthesis. Lipid accumulation increased 26% in LAM to 11.4 g/l and 18% in YPD to 4.3 g/l. Adding 1 mM and 3 mM sodium selenite to YPD medium increased total and cellular carotenoids by 22.8% (646.7 µg/L and 32.12 µg/g) and 48.7% (783.3 µg/L and 36.43 µg/g), respectively. Palmitic acid was identified as the most abundant fatty acid in all treatments, followed by oleic acid and linoleic acid. The concentrations of water soluble vitamins (WSV) and fat soluble vitamins (FSV) were generally significantly increased after supplementation with 1.0 mM sodium selenite. TEM examination revealed a significant reduction in lipid bodies accumulation in the yeast cells when sodium selenite was added to lipid-promoting environments. This decline is accompanied by an augmentation in the formation of peroxisomes, indicating that selenium has a direct impact on the degradation of fatty acids. In addition, autophagy appears to be the primary mechanism by which selenium ions are detoxified. Additionally, intracellular organelles disintegrate, cytoplasmic vacuolization occurs, and the cell wall and plasma membrane rupture, resulting in the discharge of cytoplasmic contents, when a high concentration of sodium selenite (20.0 mM) is added. Also, the presence of numerous electron-dense granules suggests an intracellular selenium-detoxification pathway. CONCLUSION: This study proposes the use of YPD with 1 mM sodium selenite to cultivate selenium-enriched biomass from R. glutinis. This approach leads to heightened lipid levels with higher accumulation of oleic, linoleic and linolenic acids, carotenoids, and vitamins. Hence, this biomass has the potential to be a valuable additive for animal, fish, and poultry feed. Furthermore, explain certain potential factors that indicate the impact of selenium in reducing the accumulation of lipid droplets in R. glutinis during lipogenesis, as detected through TEM examination.

Efficient Production of 4'-Hydroxydihydrochalcones Using Non-Conventional Yeast Strains.

The quest for novel therapeutic agents has rekindled interest in natural products, particularly those derived from biotransformation processes. Dihydrochalcones, a class of plant secondary metabolites, exhibit a range of pharmacological properties. Chalcone and dihydrochalcone compounds with the characteristic 4'-hydroxy substitution are present in 'dragon's blood' resin, known for its traditional medicinal uses and complex composition, making the isolation of these compounds challenging. This study investigates the efficient production of 4'-hydroxydihydrochalcones using non-conventional yeast strains. We evaluated the biotransformation efficiency of various 4'-hydroxychalcone substrates utilizing yeast strains such as Yarrowia lipolytica KCh 71, Saccharomyces cerevisiae KCh 464, Rhodotorula rubra KCh 4 and KCh 82, and Rhodotorula glutinis KCh 242. Our findings revealed that Yarrowia lipolytica KCh 71, Rhodotorula rubra KCh 4 and KCh 82, and Rhodotorula glutinis KCh 242 exhibited the highest conversion efficiencies, exceeding 98% within one hour for most substrates. The position of methoxy substituents in the chalcone ring significantly influenced hydrogenation efficiency. Moreover, we observed isomerization of trans-4'-hydroxy-2-methoxychalcone to its cis isomer, catalyzed by light exposure. This study underscores the potential of using yeast strains for the sustainable and efficient production of dihydrochalcones, providing a foundation for developing new therapeutic agents and nutraceuticals.

Cell wall alterations occurring in an evolved multi-stress tolerant strain of the oleaginous yeast Rhodotorula toruloides.

The oleaginous yeast species Rhodotorula toruloides is a promising candidate for applications in circular bioeconomy due to its ability to efficiently utilize diverse carbon sources being tolerant to cellular stress in bioprocessing. Previous studies including genome-wide analyses of the multi-stress tolerant strain IST536 MM15, derived through adaptive laboratory evolution from a promising IST536 strain for lipid production from sugar beet hydrolysates, suggested the occurrence of significant modifications in the cell wall. In this study, the cell wall integrity and carbohydrate composition of those strains was characterized to gain insights into the physicochemical changes associated to the remarkable multi-stress tolerance phenotype of the evolved strain. Compared to the original strain, the evolved strain exhibited a higher proportion of glucomannans, fucogalactomannans, and chitin relative to (1→4)-linked glucans, and an increased presence of glycoproteins with short glucosamine derived oligosaccharides, which have been found to be associated to ethanol stress tolerance and physical strength of the cell wall. Furthermore, the evolved strain cells were found to be significantly smaller than the original strain and more resistant to thermal and mechanical disruption, consistent with higher proportion of beta-linked polymers instead of glycogen, conferring a more rigid and robust cell wall. These findings provide further insights into the cell wall composition of this basidiomycetous red yeast species and into the alterations occurring in a multi-stress tolerant evolved strain. This new information can guide yeast genome engineering towards more robust strains of biotechnological relevance.

Mixed and membrane-separated culturing of synthetic cyanobacteria-yeast consortia reveals metabolic cross-talk mimicking natural cyanolichens.

Metabolite exchange mediates crucial interactions in microbial communities, significantly impacting global carbon and nitrogen cycling. Understanding these chemically-mediated interactions is essential for elucidating natural community functions and developing engineered synthetic communities. This study investigated membrane-separated bioreactors (mBRs) as a novel tool to identify transient metabolites and their producers/consumers in mixed microbial communities. We compared three co-culture methods (direct mixed, 2-chamber mBR, and 3-chamber mBR) to grow a synthetic binary community of the cyanobacterium Synechococcus elongatus PCC 7942 and the fungus Rhodotorula toruloides NBRC 0880, as well as axenic S. elongatus. Despite not being natural lichen constituents, these organisms exhibited interactions resembling those in cyanolichens. S. elongatus fixed CO2 into sugars as the primary shared metabolite, while R. toruloides secreted various biochemicals, predominantly sugar alcohols, mirroring the metabolite exchange observed in natural lichens. The mBR systems successfully captured metabolite gradients and revealed rapidly consumed compounds, including TCA cycle intermediates and amino acids. Our approach demonstrated that the 2-chamber mBR optimally balanced metabolite exchange and growth dynamics. This study provides insights into cross-species metabolic interactions and presents a valuable tool for investigating and engineering synthetic microbial communities with potential applications in biotechnology and environmental science.

Corn stover variability drives differences in bisabolene production by engineered Rhodotorula toruloides.

Microbial conversion of lignocellulosic biomass represents an alternative route for production of biofuels and bioproducts. While researchers have mostly focused on engineering strains such as Rhodotorula toruloides for better bisabolene production as a sustainable aviation fuel, less is known about the impact of the feedstock heterogeneity on bisabolene production. Critical material attributes like feedstock composition, nutritional content, and inhibitory compounds can all influence bioconversion. Further, the given feedstocks can have a marked influence on selection of suitable pretreatment and hydrolysis technologies, optimizing the fermentation conditions, and possibly even modifying the microorganism's metabolic pathways, to better utilize the available feedstock. This work aimed to examine and understand how variations in corn stover batches, anatomical fractions, and storage conditions impact the efficiency of bisabolene production by R. toruloides. All of these represent different facets of feedstock heterogeneity. Deacetylation, mechanical refining, and enzymatic hydrolysis of these variable feedstocks served as the basis of this research. The resulting hydrolysates were converted to bisabolene via fermentation, a sustainable aviation fuel precursor, using an engineered R. toruloides strain. This study showed that different sources of feedstock heterogeneity can influence microbial growth and product titer in counterintuitive ways, as revealed through global analysis of protein expression. The maximum bisabolene produced by R. toruloides was on the stalk fraction of corn stover hydrolysate (8.89 ± 0.47 g/L). Further, proteomics analysis comparing the protein expression between the anatomic fractions showed that proteins relating to carbohydrate metabolism, energy production, and conversion as well as inorganic ion transport metabolism were either significantly upregulated or downregulated. Specifically, downregulation of proteins related to the iron-sulfur cluster in stalk fraction suggests a coordinated response by R. toruloides to maintain overall metabolic balance, and this was corroborated by the concentration of iron in the feedstocks. ONE-SENTENCE SUMMARY: This study elucidates the effects of different sources of corn stover on bisabolene production by engineered Rhodotorula toruloides, highlighting the importance of understanding feedstock variability to enhance bioprocess efficiency and economic outcomes.

Engineering an Epoxide Hydrolase for Chemoenzymatic Asymmetric Synthesis of Chiral Triazole Fungicide (S)- and (R)-Flutriafol.

Flutriafol, a globally utilized triazole fungicide in agriculture, is typically applied as a racemic mixture, but its enantiomers differ in bioactivity and environmental impact. The synthesis of flutriafol enantiomers is critically dependent on chiral precursors: 2,2-bisaryl-substituted oxirane [(2-fluorophenyl)-2-(4-fluorophenyl)oxirane, 1a] and 1,2-diol [1-(2-fluorophenyl)-1-(4-fluorophenyl)ethane-1,2-diol, 1b]. Here, we engineered a Rhodotorula paludigensis epoxide hydrolase (RpEH), obtaining mutant Escherichia coli/RpehH336W/L360F with a 6.4-fold enhanced enantiomeric ratio (E) from 5.5 to 35.4. This enabled a gram-scale resolution of rac-1a by E. coli/RpehH336W/L360F, producing (S)-1a (98.2% ees) and (R)-1b (75.0% eep) with 44.3 and 55.7% analytical yields, respectively. As follows, chiral (S)-flutriafol (98.2% ee) and (R)-flutriafol (75.0% ee) were easily synthesized by a one-step chemocatalytic process from (S)-1a and a two-step chemocatalytic process from (R)-1b, respectively. This chemoenzymatic approach offers a superior alternative for the asymmetric synthesis of flutriafol enantiomers. Furthermore, molecular dynamics simulations revealed insight into the enantioselectivity improvement of RpEH toward bulky 2,2-bisaryl-substituted oxirane 1a.

The dynamic roles of intracellular vacuoles in heavy metal detoxification by Rhodotorula mucilaginosa.

AIMS: Rhodotorula mucilaginosa (Rho) can develop a range of strategies to resist the toxicity of heavy metals. This study aimed to investigate the physiological responses and transcriptomic regulation of the fungus under different heavy metal stresses. METHODS AND RESULTS: This study applied transmission electron microscopy and RNA-seq to investigate the fungal resistance to Pb, Cd, and Cu stresses. Under Pb stress, the activated autophagy-related genes, vesicle-fusing ATPase, and vacuolar ATP synthase improved vacuolar sequestration. This offsets the loss of lipids. However, the metal sequestration by vacuoles was not improved under Cd stress. Vacuolar fusion was also inhibited following the interference of intravacuolar Ca2+ due to their similar ionic radii. Cu2+ showed the maximum toxic effects due to its lowest cellular sorption (as low as 7%) with respect to Pb2+ and Cd2+, although the efflux pumps and divalent metal ion transporters partially contributed to the detoxification. CONCLUSIONS: Divalent cation transporters and vacuolar sequestration are the critical strategies for Rho to resist Pb stress. Superoxide dismutase (SOD) is the main strategy for Cd resistance in Rho. The intracellular Cu level was decreased by efflux pump and divalent metal ion transporters.

Engineering Rhodosporidium toruloides for sustainable production of value-added punicic acid from glucose and wood residues.

Rhodosporidium toruloides has emerged as a prominent candidate for producing single-cell oil from cost-effective feedstocks. In this study, the capability of R. toruloides to produce punicic acid (PuA), a representative plant unusual fatty acid, was investigated. The introduction of acyl lipid desaturase and conjugase (PgFADX) allowed R. toruloides to accumulate 3.7 % of total fatty acids as PuA. Delta-12 acyl lipid desaturase (PgFAD2) and diacylglycerol acyltransferase 2 were shown to benefit PuA production. The strain with PgFADX and PgFAD2 coexpression accumulated 12 % of its lipids as PuA from glucose, which translated into a PuA titer of 451.6 mg/L in shake flask condition. Utilizing wood hydrolysate as the feedstock, this strain produced 6.4 % PuA with a titer of 310 mg/L. Taken together, the results demonstrated that R. toruloides could serve as an ideal platform for the production of plant-derived high-value conjugated fatty acid using agricultural and forestry waste as feedstock.

Accumulated melanin in molds provides wavelength-dependent UV tolerance.

Fungal contamination poses a serious threat to public health and food safety because molds can grow under stressful conditions through melanin accumulation. Although ultraviolet (UV) irradiation is popular for inhibiting microorganisms, its effectiveness is limited by our insufficient knowledge about UV tolerance in melanin-accumulating molds. In this study, we first confirmed the protective effect of melanin by evaluating the UV sensitivity of young and mature spores. Additionally, we compared UV sensitivity between spores with accumulated melanin and spores prepared with melanin biosynthesis inhibitors. We found that mature spores were less UV-sensitive than young spores, and that reduced melanin accumulation by inhibitors led to reduced UV sensitivity. These results suggest that melanin protects cells against UV irradiation. To determine the most effective wavelength for inhibition, we evaluated the wavelength dependence of UV tolerance in a yeast (Rhodotorula mucilaginosa) and in molds (Aspergillus fumigatus, Cladosporium halotolerans, Cladosporium sphaerospermum, Aspergillus brasiliensis, Penicillium roqueforti, and Botrytis cinerea). We assessed UV tolerance using a UV-light emitting diode (LED) irradiation system with 13 wavelength-ranked LEDs between 250 and 365 nm, a krypton chlorine (KrCl) excimer lamp device, and a low pressure (LP) Hg lamp device. The inhibition of fungi peaked at around 270 nm, and most molds showed reduced UV sensitivity at shorter wavelengths as they accumulated pigment. Absorption spectra of the pigments showed greater absorption at shorter wavelengths, suggesting greater UV protection at these wavelengths. These results will assist in the development of fungal disinfection systems using UV, such as closed systems of air and water purification.

The potential of magnesium oxide nanoparticles (MgONPs) in the transesterification of lipids produced by Rhodotorula minuta.

The urgent need to address energy security risks and global warming has led to exploration of renewable energy sources. One such avenue is biodiesel specifically focusing on the potential of Rhodotorula minuta, a type of yeast known for producing lipids that can be used as a sustainable alternative for production of biodiesel. In the current study, this promising yeast was evaluated for its potential to produce lipids. The morphological characterization was carried out by scanning electron microscope (SEM), and intracellular detail was studied by transmission electron microscope (TEM). Changes in content and cellular biomass were monitored at time intervals with the highest biomass yield of 12.4 g/l and lipid content of 6.2 g/l achieved after 72 h. In the present work, magnesium oxide nanoparticles (MgO NPs) were synthesized and extensively characterized through Fourier transform infrared spectroscopy (FTIR), thermogravimetric analysis (TGA), Brunauer-Emmett-Teller (BET), SEM, TEM, and X-ray diffraction (XRD). By employing response surface methodology (RSM) with Box-Behnken design (BBD), optimal process conditions for transesterification could be determined. The best result achieved was a yield of 88.6% when the conditions were optimized, using methanol to oil ratio of 18:1 and 8% (w/w) amount of catalyst maintaining a reaction temperature of 55 °C and allowing the reaction to proceed for 120 min.

Clostridium carboxidivorans and Rhodosporidium toruloides as a platform for the valorization of carbon dioxide to microbial oils.

There is growing scientific interest in oleaginous yeasts producing microbial oils as precursors of biofuels and potential substitutes for fossil fuels. Due to the high cost of substrates commonly metabolized by yeasts, volatile fatty acids (VFAs) are gaining interest as alternative cheap and sustainable carbon sources, which can be obtained from solid, liquid and gas pollutants. In this research, Rhodosporidium toruloides was proven to be able to accumulate microbial oils from VFAs obtained from the fermentation of syngas by Clostridium carboxidivorans. Using CO2 and CO as carbon sources from the syngas mixture and H2 as energy source, this acetogen produced, via the Wood-Ljungdahl pathway, a mixture of acetic, butyric and caproic acids. It was first revealed that R. toruloides exhibited minimal inhibition at concentrations below 12 g/L when exposed to a mixture of VFAs, which included acetic, butyric and even hexanoic acids. The yeast was then grown on the culture medium derived from the acetogenic fermentation of syngas. Between the two yeast strains tested of the same species, R. toruloides DSM 4444 reached a total VFAs consumption of 69.1 g/L, supplied by successive additions of acids to the reactor, yielding a maximum lipid content of 29.7% w/w cell. The lipid profile obtained in this case, in terms of abundance followed the order C18:1 > C16:0 ≥ C18:0 > C18:2>others; in which the dominant compound (C18:1), represented approximately 50% of the total. This research opens new possibilities in the cultivation of oleaginous yeasts for the production of biofuels and bioproducts from C1 gases.

Isolation and Identification of Highly Sb-Resistant Rhodotorula glutinis Strain J5 and its Mechanism of Resistance to Sb(III).

Sb-resistant strains can detoxify antimony through metabolic mechanisms such as oxidation and affect the migration, transformation, and ultimate fate of antimony in the environment. In this study, a strain of Sb-resistant fungi, Rhodotorula glutinis sp. Strain J5, was isolated from Xikuangshan mine and its growth characteristics, gene expression differences, and functional annotation under Sb(III) stress were further investigated to reveal the mechanism of resistance to Sb(III). We identified strain J5 as belonging to the Rhodotorula glutinis species optimally growing at pH 5.0 and at 28 °C of temperature. According to gene annotation and differential expression, the resistance mechanism of Strain J5 includes: reducing the endocytosis of antimony by aquaporin AQP8 and transmembrane transporter pst, enhancing the efflux of Sb(III) by the gene expression of acr2, acr3 and ABC, improving the oxidation of Sb(III) by iron-sulfur protein and Superoxide dismutase (SOD), glutathione (GSH) and cysteine (Cys) chelation, methylation of methyltransferase and N-methyltransferase, accelerating cell damage repair and EPS synthesis and other biochemical reaction mechanisms. FT-IR analysis shows that the -OH, -COOH, -NH, -PO, C-O, and other active groups of Strain J5 can be complexed with Sb(III), resulting in chemical adsorption. Strain J5 displays significant resistance to Sb(III) with the MIC of 1300 mg/L, playing a crucial role in the global biochemical transformation of antimony and its potential application in soil microbial remediation.

Engineering oleaginous red yeasts as versatile chassis for the production of oleochemicals and valuable compounds: Current advances and perspectives.

Enabling the transition towards a future circular bioeconomy based on industrial biomanufacturing necessitates the development of efficient and versatile microbial platforms for sustainable chemical and fuel production. Recently, there has been growing interest in engineering non-model microbes as superior biomanufacturing platforms due to their broad substrate range and high resistance to stress conditions. Among these non-conventional microbes, red yeasts belonging to the genus Rhodotorula have emerged as promising industrial chassis for the production of specialty chemicals such as oleochemicals, organic acids, fatty acid derivatives, terpenoids, and other valuable compounds. Advancements in genetic and metabolic engineering techniques, coupled with systems biology analysis, have significantly enhanced the production capacity of red yeasts. These developments have also expanded the range of substrates and products that can be utilized or synthesized by these yeast species. This review comprehensively examines the current efforts and recent progress made in red yeast research. It encompasses the exploration of available substrates, systems analysis using multi-omics data, establishment of genome-scale models, development of efficient molecular tools, identification of genetic elements, and engineering approaches for the production of various industrially relevant bioproducts. Furthermore, strategies to improve substrate conversion and product formation both with systematic and synthetic biology approaches are discussed, along with future directions and perspectives in improving red yeasts as more versatile biotechnological chassis in contributing to a circular bioeconomy. The review aims to provide insights and directions for further research in this rapidly evolving field. Ultimately, harnessing the capabilities of red yeasts will play a crucial role in paving the way towards next-generation sustainable bioeconomy.

Chitosan-coated manganese ferrite nanoparticles enhanced Rhodotorula toruloides carotenoid production.

The present study aims to analyze the interaction between Rhodotorula toruloides and magnetic nanoparticles and evaluate their effect on carotenoid production. The manganese ferrite nanoparticles were synthesized without chitosan (MnFe2O4) and chitosan coating (MnFe2O4-CS) by the co-precipitation method assisted by hydrothermal treatment. XRD (X-ray diffraction), Magnetometry, Dynamic Light Scattering (DLS) and FTIR (Fourier-Transform Infrared Spectroscopy), are used to characterize the magnetic nanoparticles. The crystallite size of MnFe2O4 was 16 nm for MnFe2O4 and 20 nm for MnFe2O4-CS. The magnetic saturation of MnFe2O4-CS was lower (39.6 ± 0.6 emu/g) than the same MnFe2O4 nanoparticles (42.7 ± 0.3 emu/g), which was attributed to the chitosan fraction presence. The MnFe2O4-CS FTIR spectra revealed the presence of the characteristic chitosan bands. DLS demonstrated that the average hydrodynamic diameters were 344 nm for MnFe2O4 and 167 nm for MnFe2O4-CS. A kinetic study of cell immobilization performed with their precipitation with a magnet demonstrated that interaction between magnetic nanoparticles and R. toruloides was characterized by an equilibrium time of 2 h. The adsorption isotherm models (Langmuir and Freundlich) were fitted to the experimental values. The trypan blue assay was used for cell viability assessment. The carotenoid production increased to 256.2 ± 6.1 µg/g dry mass at 2.0 mg/mL MnFe2O4-CS. The use of MnFe2O4-CS to stimulate carotenoid yeast production and the magnetic separation of biomass are promising nanobiotechnological alternatives. Magnetic cell immobilization is a perspective technique for obtaining cell metabolites.

[Screening and fermentation of high-yield glycolic acid strains].

Glycolic acid is an important chemical product widely used in various fields, including cosmetics, detergents, textiles, and more. Currently, microbial production of glycolic acid has disadvantages such as poor genetic stability, low yield, and high cost. Additionally, whole-cell catalytic production of glycolic acid typically requires the addition of relatively expensive sorbitol as a carbon source, which limits its industrial production. To develop an industrially applicable method for glycolic acid production, this study used ethylene glycol as a substrate to screen the glycolic acid-producing strains through whole-cell catalysis, obtaining a Rhodotorula sp. capable of producing glycolic acid. The strain was then subjected to UV mutagenesis and high throughput screening, and the positive mutant strain RMGly-20 was obtained. After optimization in shake flasks, the glycolic acid titer of RMGly-20 reached 17.8 g/L, a 10.1-fold increase compared to the original strain. Using glucose as the carbon source and employing a fed-batch culture in a 5 L fermenter, strain RMGly-20 produced 61.1 g/L of the glycolic acid. This achievement marks the preliminary breeding of a genetically stable glycolic acid-producing strain using a cheap carbon source, providing a new host for the biosynthesis of glycolic acid and promoting further progress toward industrial production.

Optimized phenol degradation and lipid production by Rhodosporidium toruloides using response surface methodology and genetic algorithm-optimized artificial neural network.

Oleaginous yeast can produce lipids while degrading phenol in wastewater treatment. In this study, a Plackett-Burman Design (PBD) was adopted to identify key factors of phenol degradation and lipid production using R toruloides 9564T. While temperature, inoculum size, and agitation were significant for both the processes (p < 0.05), pH and incubation were significant for lipid production, and phenol removal, respectively. Results from four factors (pH, temperature, inoculum size, and incubation period) central composite design (CCD) experiment were used to formulate quadratic and genetic algorithm-optimized ANN models. The reduced quadratic model for phenol degradation (R2: 0.993) and lipid production (R2: 0.958) were marginally inferior to ANN models (R2: 0.999, 0.982, respectively) on training sets. Multi-objective optimization with equal importance suggests phenol degradation between 106.4 and 108.76%, and lipid production of 0.864-0.903 g/L, by polynomial and ANN models. Complete phenol degradation (100%) and 3.35-fold increment (0.918 g/L) in lipid production were obtained at pH 6.07, inoculum size 14.68% v/v, at 29.5 °C in 92.17 h experimentally.

Efficient sugar utilization and high tolerance to inhibitors enable Rhodotorula toruloides C23 to robustly produce lipid and carotenoid from lignocellulosic feedstock.

The utilization of lignocellulosic substrates for microbial oil production by oleaginous yeasts has been evidenced as an economically viable process for industrial-scale biodiesel preparation. Efficient sugar utilization and tolerance to inhibitors are critical for lipid production from lignocellulosic substrates. This study investigated the lignocellulosic sugar utilization and inhibitor tolerance characteristics of Rhodotorula toruloides C23. The results demonstrated that C23 exhibited robust glucose and xylose assimilation irrespective of their ratios, yielding over 21 g/L of lipids and 11 mg/L of carotenoids. Furthermore, C23 exhibited high resistance and efficiently degradation towards toxic inhibitors commonly found in lignocellulosic hydrolysates. The potential molecular mechanism underlying xylose metabolism in C23 was explored, with several key enzymes and signal regulation pathways identified as potentially contributing to its superior lipid synthesis performance. The study highlights R. toruloides C23 as a promising candidate for robust biofuel and carotenoid production through direct utilization of non-detoxified lignocellulosic hydrolysates.

Comparative genomic analysis and optimization of astaxanthin production of Rhodotorula paludigena TL35-5 and Rhodotorula sampaioana PL61-2.

Astaxanthin is a powerful antioxidant known to enhance skin, cardiovascular, eye, and brain health. In this study, the genome insights and astaxanthin production of two newly isolated astaxanthin-producing yeasts (TL35-5 and PL61-2) were evaluated and compared. Based on their phenotypic and genotypic characteristics, TL35-5 and PL61-2 were identified as basidiomycetous yeasts belonging to Rhodotorula paludigena and Rhodotorula sampaioana, respectively. To optimize astaxanthin production, the effects of cultural medium composition and cultivation conditions were examined. The optimal conditions for astaxanthin production in R. paludigena TL35-5 involved cultivation in AP medium containing 10 g/L glucose as the sole carbon source, supplemented with 1.92 g/L potassium nitrate, pH 6.5, and incubation at 20°C for 3 days with shaking at 200 rpm. For R. sampaioana PL61-2, the optimal medium composition for astaxanthin production consisted of AP medium with 40 g/L glucose, supplemented with 0.67 g/L urea, pH 7.5, and the fermentation was carried out at 20°C for 3 days with agitating at 200 rpm. Under their optimal conditions, R. paludigena TL35-5 and R. sampaioana PL61-2 gave the highest astaxanthin yields of 3.689 ± 0.031 and 4.680 ± 0.019 mg/L, respectively. The genome of TL35-5 was 20,982,417 bp in length, with a GC content of 64.20%. A total of 6,789 protein-encoding genes were predicted. Similarly, the genome of PL61-2 was 21,374,169 bp long, with a GC content of 64.88%. It contained 6,802 predicted protein-encoding genes. Furthermore, all essential genes involved in astaxanthin biosynthesis, including CrtE, CrtYB, CrtI, CrtS, and CrtR, were identified in both R. paludigena TL35-5 and R. sampaioana PL61-2, providing evidence for their ability to produce astaxanthin.

Bioreduction of 4'-Hydroxychalcone in Deep Eutectic Solvents: Optimization and Efficacy with Various Yeast Strains.

4'-dihydrochalcones are secondary metabolites isolated from many medicinal plants and from the resin known as 'dragon's blood'. Due to their biological potential, our research objective was to determine the possibilities of using biocatalysis processes carried out in deep eutectic solvents (DESs) to obtain 4'-dihydrochalcones as a model compound. The processes were carried out in a culture of the yeast Yarrowia lipolytica KCh 71 and also in cultures of strains of the genera Rhodotorula and Debaryomyces. Based on the experiments carried out, an optimum process temperature of 35 °C was chosen, and the most suitable DES contained glycerol as a hydrogen bond donor (HBD). For a medium with 30% water content (DES 11), the conversion observed after 24 h exceeded 70%, while increasing the amount of water to 50% resulted in a similar level of conversion after just 1 h. A fivefold increase in the amount of added substrate resulted in a reduction in conversion, which reached 30.3%. Of the other yeast strains tested, Rhodotorula marina KCh 77 and Rhodotorula rubra KCh 4 also proved to be good biocatalysts for the bioreduction process. For these strains, the conversion reached 95.4% and 95.1%, respectively. These findings highlight the potential of yeast as a biocatalyst for the selective reduction of α,ß-unsaturated ketones and the possibility of using a DESs as a reaction medium in this process.

Technological modes and processes to enhance the Rhodosporidium toruloides based lipid accumulation.

Rhodosporidium toruloides has emerged as an excellent option for microbial lipid production due to its ability to accumulate up to 70 % of lipids per cell dry weight, consume multiple substrates such as glucose and xylose, and tolerate toxic compounds. Despite the potential of Rhodosporidium toruloides for high lipid yields, achieving these remains is a significant hurdle. A comprehensive review is essential to thoroughly evaluate the advancements in processes and technologies to enhance lipid production in R. toruloides. The review covers various strategies for enhancing lipid production like co-culture, adaptive evolution, carbon flux analysis, as well as different modes of fermentation. This review will help researchers to better understand the recent developments in technologies for sustainable and scalable lipid production from R. toruloides and simultaneously emphasize the need for developing an efficient and sustainable bioprocess.