Identification of Irpex and Rhodotorula on surveillance bronchoscopy in a pediatric lung transplant recipient: A case report and review of literature of these atypical fungal organisms.

BACKGROUND: Invasive fungal disease (IFD) is a frequent complication in pediatric lung transplant recipients, occurring in up to 12% of patients in the first year. Risk factors for infection include impaired lung defenses and intense immunosuppressive regimens. While most IFD occurs from Aspergillus, other fungal conidia are continuously inhaled, and infections with fungi on a spectrum of human pathogenicity can occur. CASE REPORT: We report a case of a 17-year-old lung transplant recipient in whom Irpex lacteus and Rhodotorula species were identified during surveillance bronchoscopy. She was asymptomatic and deemed to be colonized by Irpex lacteus and Rhodotorula species following transplant. 2 years after transplantation, she developed a fever, respiratory symptoms, abnormal lung imaging, and histological evidence of acute and chronic bronchitis on transbronchial biopsy. After developing symptoms concerning for a pulmonary infection and graft dysfunction, she was treated for a presumed IFD. Unfortunately, further diagnostic testing could not be performed at this time given her tenuous clinical status. Despite the initiation of antifungal therapy, her graft function continued to decline resulting in a second lung transplantation. CONCLUSIONS: This case raises the concern for IFD in lung transplant recipients from Irpex species. Further investigation is needed to understand the pathogenicity of this organism, reduce the incidence and mortality of IFD in lung transplant recipients, and refine the approach to diagnosis and manage the colonization and isolation of rare, atypical fungal pathogens in immunocompromised hosts.

Carotenoids production by Rhodosporidium paludigenum yeasts: Characterization of chemical composition, antioxidant and antimicrobial properties.

The high market potential imposed by natural carotenoids has turned the scientific interest in search for new strains, capable of synthesizing a wide spectrum of these pigments. In this study, Rhodosporidium paludigenum NCYC 2663 and 2664 were investigated for carotenoids production and lipid accumulation utilizing different carbon sources (glucose, fructose, sucrose, mixture of glucose: galactose). Strain R. paludigenum 2663 produced the highest total carotenoids titer (2.21 mg/L) when cultivated on sucrose, together with 4 g/L lipids (30% w/w content) and 7 g/L exopolysaccharides. In the case of R. paludigenum 2664, glucose favored the production of 2.93 mg/L total carotenoids and 1.57 g/L lipids (31.8% w/w content). Analysis of the chemical profile during fermentation revealed that ß-carotene was the prominent carotenoid. Strain 2663 co-produced γ-carotene, torulene and torularhodin in lower amounts, whereas 2664 synthesized almost exclusively ß-carotene. The produced lipids from strain 2663 were rich in oleic acid, while the presence of linoleic acid was also detected in the lipoic fraction from strain 2664. The obtained carotenoid extracts exhibited antioxidant (IC50 0.14 mg/mL) and high antimicrobial activity, against common bacterial and fungal pathogenic strains. The results of this study are promising for the utilization of biotechnologically produced carotenoids in food applications.

Diversity, distribution, and bioprospecting potentials of carotenogenic yeast from mangrove ecosystem.

Microbial production of carotenoids has gained significant interest for its cost-effectiveness and sustainable nature. This study focuses on 47 red-pigmented yeasts isolated from sediments and plant parts of 13 species of mangrove trees. The relative abundance and distribution of these yeasts varied with plant species and plant parts. The highest number of red yeasts was associated with the mangrove plant Avicennia officinalis (32%). Notably, the leaves harbored the highest percentage (45%) of carotenogenic yeasts, and definite compartmentalization of these yeast species was noticed in mangrove plant parts. All the isolates were molecularly identified and they belonged to the genera of Rhodotorula, Rhodosporidiobolus, and Cryptococcus. The diversity of the pigmented yeasts isolated from A. officinalis was found to be the greatest. Among these strains, Rhodotorula mucilaginosa PV 8 was identified as the most potent producer of carotenoid pigment. Under optimized conditions of physical parameters - 28 °C, pH 5, and 15% salinity led to biomass production of 9.2 ± 0.12 g/L DCW and a pigment yield of 194.78 µg/g. The pigment produced by PV 8 was identified as ß-carotene by thin layer chromatography (TLC) and Fourier transform infrared spectroscopy (FT-IR). This ß-carotene demonstrated strong antioxidant activity. Moreover, the carotenoid displayed promising antibacterial activity against multidrug-resistant organisms, including Aeromonas sp. and Vibrio sp. In vitro studies revealed the probiotic traits of PV 8. The cytotoxicity of R. mucilaginosa PV 8 was assessed in the invertebrate model Artemia salina and the survival rate showed that it was non-toxic. Furthermore, the ß-carotene from PV 8 demonstrated the ability to transfer its vibrant color to various food products, maintaining color stability even under varied conditions. This research underscores the potential of R. mucilaginosa PV 8, as a versatile and valuable resource for the production of carotenoids.

Glucosylglycerol and proline reverse the effects of glucose on Rhodosporidium toruloides lifespan.

Rhodosporidium toruloides is a novel cell factory used to synthesis carotenoids, biosurfactants, and biofuel feedstocks. However, research on R. toruloides has generally centred on the manufacture of biochemicals, while analyses of its longevity have received scant attention. Understanding of R. toruloides longevity under different nutrient conditions could help to improve its biotechnological significance and metabolite production. Glucosylglycerol (GG) and proline are osmoprotectants that could revert the harmful effects of environmental stress. This study examined how GG and proline affect R. toruloides strain longevity under glucose nutrimental stress. Herein, we provide evidence that GG and proline enhance cell performance and viability. These compatible solutes neutralises the pro-ageing effects of high glucose (10% glucose) on the yeast cell and reverse its cellular stress. GG exhibits the greatest impact on lifespan extension at 100 mM, whereas proline exerts effect at 2 mM. Our data reveal that these compounds significantly affect the culture medium osmolarity. Moreso, GG and proline decreased ROS production and mitohormetic lifespan regulation, respectively. The data indicates that these solutes (proline and GG) support the longevity of R. toruloides at a pro-ageing high glucose culture condition.

Formation of Sb2O3 microcrystals by Rhodotorula mucilaginosa.

Antimony (Sb) pollution seriously endangers ecological environment and human health. Microbial induced mineralization can effectively convert metal ions into more stable and less soluble crystalline minerals by extracellular polymeric substance (EPS). In this study, an efficient Sb-resistant Rhodotorula mucilaginosa (R. mucilaginosa) was screened, which can resist 41 mM Sb(III) and directly transform Sb(III) into Sb2O3 microcrystals by EPS. The removal efficiency of R. mucilaginosa for 22 mM Sb(III) reached 70% by converting Sb(III) to Sb2O3. The components of supernatants as well as the effects of supernatants and pH on Sb(III) mineralization verified that inducible and non-inducible extracellular protein/polysaccharide biomacromolecules play important roles in the morphologies and sizes control of Sb2O3 formed by R. mucilaginosa respectively. Sb2O3 microcrystals with different morphologies and sizes can be prepared by the regulation of inducible and non-inducible extracellular biomacromolecules secreted by R. mucilaginosa. This is the first time to identify that R. mucilaginosa can remove Sb(III) by transforming Sb(III) into Sb2O3 microcrystals under the control of EPS. This study contributes to our understanding for Sb(III) biomineralization mechanisms and provides strategies for the remediation of Sb-contaminated environment.

The mitochondrial respiratory chain from Rhodotorula mucilaginosa, an extremophile yeast.

Rhodotorula mucilaginosa survives extreme conditions through several mechanisms, among them its carotenoid production and its branched mitochondrial respiratory chain (RC). Here, the branched RC composition was analyzed by biochemical and complexome profiling approaches. Expression of the different RC components varied depending on the growth phase and the carbon source present in the medium. R. mucilaginosa RC is constituted by all four orthodox respiratory complexes (CI to CIV) plus several alternative oxidoreductases, in particular two type-II NADH dehydrogenases (NDH2) and one alternative oxidase (AOX). Unlike others, in this yeast the activities of the orthodox and alternative respiratory complexes decreased in the stationary phase. We propose that the branched RC adaptability is an important factor for survival in extreme environmental conditions; thus, contributing to the exceptional resilience of R. mucilaginosa.

Production optimization of food functional factor ergothioneine in wild-type red yeast Rhodotorula mucilaginosa DL-X01.

BACKGROUND: Ergothioneine (EGT) is a high-value food functional factor that cannot be synthesized by humans and other vertebrates, and the low yield limits its application. RESULTS: In this study, the optimal fermentation temperature, fermentation time, initial pH, inoculum age, and inoculation ratio on EGT biosynthesis of Rhodotorula mucilaginosa DL-X01 were optimized. In addition, the effects of three key precursor substances - histidine, methionine, and cysteine - on fungal EGT synthesis were verified. The optimal conditions were further obtained by response surface optimization. The EGT yield of R. mucilaginosa DL-X01 under optimal fermentation conditions reached 64.48 ± 2.30 mg L-1 at shake flask fermentation level. Finally, the yield was increased to 339.08 ± 3.31 mg L-1 (intracellular) by fed-batch fermentation in a 5 L bioreactor. CONCLUSION: To the best of our knowledge, this is the highest EGT yield ever reported in non-recombinant strains. The fermentation strategy described in this study will promote the efficient biosynthesis of EGT in red yeast and its sustainable production in the food industry. © 2024 Society of Chemical Industry.

Transcriptomic Analysis Reveals the Potential Mechanisms for Improving Carotenoid Production in Rhodosporidium toruloides Z11 under Light Stress.

Carotenoids, as a type of tetraterpene compound, have been widely used in food, medical, and health areas owing to their antioxidant, immune enhancement, and disease risk reduction effects. Rhodosporidium toruloides is a promising oleaginous red yeast that can industrially synthesize carotenoids. In this study, the effects of different light exposure times and intervals on carotenoid production by R. toruloides Z11 were first investigated. Results showed that a higher carotenoid content (1.29 mg/g) can be achieved when R. toruloides Z11 was exposed to light for 12 h per day, which was increased by 1.98 times compared with that of dark cultivation. Transcriptome profiling revealed that light stress could effectively promote the gene expression levels of GGPS1 and AL1 in the carotenoid biosynthesis pathway and phr in the DNA photolysis pathway of R. toruloides. This work will provide a molecular foundation to further improve the production efficiency of carotenoids by genetic engineering.

Effect of Rhodotorula mucilaginosa inoculation on the aroma development of a fermented vegetables simulated system.

Fermented vegetables are known for their unique flavors and aromas, which are influenced by the complex microbial processes that occur during fermentation. Rhodotorula mucilaginosa is a red yeast strain that is frequently isolated from fermented vegetables. However, the specific mechanisms underlying their effects on aroma production remain unclear. In this study, a simulated system of vegetables fermented using vegetable juices was used to investigate the effects of R. mucilaginosa inoculation on aroma development. The results demonstrated that this red yeast strain could utilize the nutrients present in the vegetable juices to support its growth and reproduction. Moreover, the inoculation of fermented vegetable juices with this yeast strain led to an increase in the levels of umami amino acids and sweet amino acids. Furthermore, this yeast strain was found able to significantly reduce the content of sulfur-containing compounds, which may decrease the unpleasant odor of fermented vegetables. Additionally, the yeast strain was capable of producing high concentrations of aromatic compounds such as phenylethyl alcohol, methyl 2-methylbutyrate, methyl butyrate, and nonanoic acid in a minimum medium. However, only phenylethyl alcohol has been identified as a core aromatic compound in fermented vegetable juice. The three fermented vegetable juices exhibited significantly different flavor profiles according to comparative analysis. Therefore, the core flavor compounds found in fermented vegetables are primarily derived from the release and modification of endogenous flavors naturally present in the vegetables, facilitated by the yeast during fermentation.

Research progress on carotenoid production by Rhodosporidium toruloides.

Carotenoids are natural lipophilic pigments, which have been proven to provide significant health benefits to humans, relying on their capacity to efficiently scavenge singlet oxygen and peroxyl radicals as antioxidants. Strains belonging to the genus Rhodosporidium represent a heterogeneous group known for a number of phenotypic traits including accumulation of carotenoids and lipids and tolerance to heavy metals and oxidative stress. As a representative of these yeasts, Rhodosporidium toruloides naturally produces carotenoids with high antioxidant activity and grows on a wide variety of carbon sources. As a result, R. toruloides is a promising host for the efficient production of more value-added lipophilic compound carotenoids, e.g., torulene and torularhodin. This review provides a comprehensive summary of the research progress on carotenoid biosynthesis in R. toruloides, focusing on the understanding of biosynthetic pathways and the regulation of key enzymes and genes involved in the process. Moreover, the relationship between the accumulation of carotenoids and lipid biosynthesis, as well as the stress from diverse abiotic factors, has also been discussed for the first time. Finally, several feasible strategies have been proposed to promote carotenoid production by R. toruloides. It is possible that R. toruloides may become a critical strain in the production of carotenoids or high-value terpenoids by genetic technologies and optimal fermentation processes. KEY POINTS: • Biosynthetic pathway and its regulation of carotenoids in Rhodosporidium toruloides were concluded • Stimulation of abiotic factors for carotenoid biosynthesis in R. toruloides was summarized • Feasible strategies for increasing carotenoid production by R. toruloides were proposed.

Detoxification Approaches of Bagasse Pith Hydrolysate Affecting Xylitol Production by Rhodotorula mucilaginosa.

In this study, the potential of bagasse pith (the waste of sugar and paper industry) was investigated for bio-xylitol production for the first time. Xylose-rich hydrolysate was prepared using 8% dilute sulfuric acid, at 120 °C for 90 min. Then, the acid-hydrolyzed solution was detoxified by individual overliming (OL), active carbon (AC), and their combination (OL+AC). The amounts of reducing sugars and inhibitors (furfural and hydroxyl methyl furfural) were measured after acid pre-treatment and detoxification process. Thereafter, xylitol was produced from detoxified hydrolysate by Rhodotorula mucilaginosa yeast. Results showed that after acid hydrolysis, the sugar yield was 20%. Detoxification by overliming and active carbon methods increased the reducing sugar content up to 65% and 36% and decreased the concentration of inhibitors to >90% and 16%, respectively. Also, combined detoxification caused an increase in the reducing sugar content (>73%) and a complete removal of inhibitors. The highest productivity of xylitol (0.366 g/g) by yeast was attained after the addition of 100 g/l non-detoxified xylose-rich hydrolysate into fermentation broth after 96 h, while the xylitol productivity enhanced to 0.496 g/g after adding the similar amount of xylose-rich hydrolysate detoxified by combined method (OL+AC2.5%).

Computational assessment of lipid production in Rhodosporidium toruloides in two-stage and one-stage batch bioprocesses.

Oleaginous yeasts are promising platforms for microbial lipids production as a renewable and sustainable alternative to vegetable oils in biodiesel production. In this paper, a thorough in silico assessment of lipid production in batch cultivation by Rhodosporidium toruloides was developed. By means of dynamic flux balance analysis, the traditional two-stage bioprocess (TSB) performed by the native strain was contrasted with one-stage bioprocess (OSB) using four designed strains obtained by gene knockout strategies. Lipid titer, yield, content, and productivity were analyzed at different initial C/N ratios as relevant performance indicators used in bioprocesses. By weighting these indicators, a global lipid efficiency metric (GLEM) was defined to consider different scenarios. Under simulated conditions, designed strains for lipid overproduction in OSB outperformed the TSB in terms of lipid title (up to threefold), lipid yield (up to 2.4-fold), lipid content (up to 2.8-fold, with a maximum of 76%), and productivity (up to 1.3-fold), depending on C/N ratios. Using these efficiency parameters and the proposed GLEM, the process of selecting the most suitable candidates for lipid production could be carried out before experimental assays. This methodology holds the potential to be extended to other oleaginous microorganisms and diverse strain design techniques.

Application of adaptive laboratory evolution to improve the tolerance of Rhodotorula strain to methanol in crude glycerol and development of an effective method for cell lysis.

Rhodotorula toruloides can utilize crude glycerol as the low-cost carbon source for lipid production, but its growth is subjected to inhibition by methanol in crude glycerol. Here, transcriptome profiling demonstrated that 1004 genes were significantly regulated in the strain R. toruloides TO2 under methanol stress. Methanol impaired the function of membrane transport and subsequently weakened the utilization of glycerol, activities of the primary metabolism and functions of nucleus and ribosome. Afterwards the tolerance of TO2 to methanol was improved by using two-round adaptive laboratory evolution (ALE). The final strain M2-ale had tolerance up to 3.5% of methanol. 1 H NMR-based metabolome analysis indicated that ALE not only improved the tolerance of M2-ale to methanol but also tuned the carbon flux towards the biosynthesis of glycerolipid-related metabolites. The biomass and lipid titer of M2-ale reached 14.63 ± 0.45 g L-1 and 7.06 ± 0.44 g L-1 at 96 h in the crude glycerol medium, which increased up to 17.69% and 31.39%, respectively, comparing with TO2. Afterwards, an effective method for cell lysis was developed by combining sonication and enzymatic hydrolysis (So-EnH). The lytic effect of So-EnH was validated by using confocal imaging and flow cytometry. At last, lipid recovery rate reached 95.4 ± 2.7% at the optimized condition.

Cost-effective production of ergothioneine using Rhodotorula mucilaginosa DL-X01 from molasses and fish bone meal enzymatic hydrolysate.

Ergothioneine (EGT) is a high-value natural antioxidant that cannot be synthesized by the human body. This study showed that Rhodotorula mucilaginosa DL-X01 can use untreated molasses and fish bone meal enzymatic hydrolysate as the substrates to synthesize EGT. By optimizing the growth conditions, the EGT yield reached 29.39 mg/L when molasses and fish bone meal (FBM) were added at 60 g/L and 400 g/L respectively. Finally, the EGT yield was increased to 216.25 mg/L by fed-batch fermentation in a 5 L bioreactor. Compared with the fermentation by yeast extract peptone dextrose medium, the feedstock cost of EGT production was reduced by 330.91 % by using molasses and FBM as substrates. These results showed that R. mucilaginosa DL-X01 can produce high-value EGT using two cheap processing by-products, molasses and FBM, which is of great significance for environmental protection and sustainable development.

Environmental assessment of Rhodosporidium toruloides-1588 based oil production using wood hydrolysate and crude glycerol.

Rhodosporidium toruloides, an oleaginous yeast, is a potential feedstock for biodiesel production due to its ability to utilize lignocellulosic biomass-derived hydrolysate with a considerably high lipid titer of 50-70 % w/w. Hence, for the first-time environmental assessment of large-scale R. toruloides-based biodiesel production from wood hydrolysate and crude glycerol was conducted. The global warming potential was observed to be 0.67 kg CO2 eq./MJ along with terrestrial ecotoxicity of 1.37 kg 1,4-DCB eq./MJ and fossil depletion of 0.13 kg oil eq./MJ. The highest impacts for global warming (∼45 %) and fossil depletion (∼37 %) are attributed to the use of chloroform for lipid extraction while fuel consumption for transportation contributed more than 50 % to terrestrial ecotoxicity. Further, sensitivity analysis revealed that maximizing biodiesel yield by increasing lipid yield and solid loading could contribute to reduced environmental impacts. In nutshell, this investigation reveals that environmental impact varies with the type of chemical utilized.

Sustainable trehalose lipid production by Rhodotorula sp.: a promising bio-based alternative.

Global environmental concerns drive research toward the development of new eco-friendly compounds to replace pollutant chemicals. This study focuses on optimizing the production of trehalose lipids (TLs), which are glycolipid biosurfactants (BS) with various applications like antimicrobial or surface tension reduction. New microorganism sources, growth conditions, medium composition, purification conditions, and physicochemical properties of TLs are studied. Addressing a microscale approach, TLs production was successfully achieved using Rhodotorula sp. and Rhodococcus erythropolis to compare, with different media compositions including glucose-based and salt media supplemented with glycerol, glucose, n-hexadecane, n-dodecane. Liquid-liquid extraction using ethyl acetate and methanol was employed for compound extraction, followed by characterization using analytical methods such as Thin layer chromatography (TLC), High performance liquid chromatography (HPLC), and UHPLC. The produced TLs exhibited a minimum surface tension of 47 mN/m and a critical micellar concentration of 4.4 mg/mL. This study also identified Rhodotorula sp. as a new sustainable producer of TLs with improved productivity.

Whole-genome sequence and characterization of a marine red yeast, Rhodosporidium sphaerocarpum GDMCC 60679, featuring the assimilation of ammonia nitrogen.

A marine red yeast, Rhodosporidium sphaerocarpum, is generally used for the production of lipids and carotenoids. In a previous study, we demonstrated that a marine-derived R. sphaerocarpum GDMCC 60679 can efficiently remove ammonia nitrogen and exhibit multiple probiotic functions for shrimp, Litopenaeus vannamei. Here, we performed a genome assembly of the strain GDMCC 60679 using a combination of the data from Illumina PE and PacBio CLR reads. The genome has a size of 18.03 Mb and consists of 32 contigs with an N50 length of 1,074,774 bp and GC content of 63 %. The genome was predicted to contain 6092 protein-coding genes, 5962 of which were functionally annotated. Metabolic pathways responsible for the ammonia assimilation and the synthesis of lipids and carotenoids were particularly examined to explore and characterize genes contributing to these functions. Whole-genome sequence and annotation of the strain lays a foundation to reveal the molecular mechanism of its prominent biological functions and will facilitate us to further expand new applications of yeasts in Rhodosporidium.

Enhancement of novel Endo-polygalacturonase expression in Rhodotorula mucilaginosa PY18: insights from mutagenesis and molecular docking.

Pectinase is a particular type of enzyme that can break down pectin compounds and is extensively utilised in the agricultural field. In this study, twenty yeast isolates were isolated and assayed for pectinase activity. Molecular identification by PCR amplification and sequencing of internal transcribed spacer (ITS) regions of isolate no. 18 had the highest pectinase activity of 46.35 U/mg, was identified as Rhodotorula mucilaginosa PY18, and was submitted under accession no. (OM275426) in NCBI. Rhodotorula mucilaginosa PY18 was further enhanced through sequential mutagenesis, resulting in a mutant designated as Rhodotorula mucilaginosa E54 with a specific activity of 114.2 U/mg. Using Response Surface Methodology (RSM), the best culture conditions for the pectinase-producing yeast mutant Rhodotorula mucilaginosa E54 were pH 5, 72-h incubation, 2.5% xylose, and 2.5% malt extract, with a pectinase-specific activity of 156.55 U/mg. Then, the obtained sequences of the endo-polygalacturonase PGI gene from Rhodotorula mucilaginosa PY18 and mutant Rhodotorula mucilaginosa E54 were isolated for the first time, sequenced, and submitted to NCBI accession numbers OQ283005 and OQ283006, respectively. The modelled 3D structure of the endo-PGI enzyme (485 residues) was validated using Ramachandran's plot, which showed 87.71, 85.56, and 91.57% in the most favourable region for template Rhodotorula mucilaginosa KR, strain Rhodotorula mucilaginosa PY18, and mutant Rhodotorula mucilaginosa E54, respectively. In molecular docking studies, the results of template Rhodotorula mucilaginosa KR endo-PG1 showed an interaction with an affinity score of - 6.0, - 5.9, and - 5.6 kcal/mol for active sites 1, 2, and 3, respectively. Rhodotorula mucilaginosa PY18 endo-PG1 showed an interaction affinity with a score of - 5.8, - 6.0, and - 5.0 kcal/mol for active sites 1, 2, and 3, respectively. Mutant Rhodotorula mucilaginosa E54 endo-PG1 showed an interaction affinity of - 5.6, - 5.5, - 5.5 and - 5.4 kcal/mol for active sites 1, 2, and 3, respectively. The endo-PGI genes of both the yeast strain Rhodotorula mucilaginosa PY18 and mutant Rhodotorula mucilaginosa E54 were successfully cloned and expressed in E. coli DH5α, showing significantly higher endo-PG1 activity, which recorded 94.57 and 153.10 U/mg for recombinant Rhodotorula mucilaginosa pGEM-PGI-PY18 and recombinant mutant Rhotorula pGEM-PGI-E54, respectively.

Simultaneous Lipid and Carotenoid Production via Rhodotorula paludigena CM33 Using Crude Glycerol as the Main Substrate: Pilot-Scale Experiments.

Rhodotorula paludigena CM33 is an oleaginous yeast that has been demonstrated to accumulate substantial quantities of intracellular lipids and carotenoids. In this study, crude glycerol, a by-product of biodiesel production, was used as a carbon source to enhance the accumulation of lipids and carotenoids in the cells. The culture conditions were first optimized using response surface methodology, which revealed that the carotenoid concentration and lipid content improved when the concentration of crude glycerol was 40 g/L. Different fermentation conditions were also investigated: batch, repeated-batch, and fed-batch conditions in a 500 L fermenter. For fed-batch fermentation, the maximum concentrations of biomass, lipids, and carotenoids obtained were 46.32 g/L, 37.65%, and 713.80 mg/L, respectively. A chemical-free carotenoid extraction method was also optimized using high-pressure homogenization and a microfluidizer device. The carotenoids were found to be mostly beta-carotene, which was confirmed by HPLC (high pressure liquid chromatography), LC-MS (liquid chromatography-mass spectrometry), and NMR (nuclear magnetic resonance). The results of this study indicate that crude glycerol can be used as a substrate to produce carotenoids, resulting in enhanced value of this biodiesel by-product.

Genomic analysis of the marine yeast Rhodotorula sphaerocarpa ETNP2018 reveals adaptation to the open ocean.

BACKGROUND: Despite a rising interest in the diversity and ecology of fungi in marine environments, there are few published genomes of fungi isolated from the ocean. The basidiomycetous yeast (unicellular fungus) genus Rhodotorula are prevalent and abundant in the open ocean, and they have been isolated from a wide range of other environments. Many of these environments are nutrient poor, such as the Antarctica and the Atacama deserts, raising the question as to how Rhodotorula yeasts may have adapted their metabolic strategies to optimize survival under low nutrient conditions. In order to understand their adaptive strategies in the ocean, the genome of R. sphaerocarpa ETNP2018 was compared to that of fourteen representative Rhodotorula yeasts, isolated from a variety of environments. RESULTS: Rhodotorula sphaerocarpa ETNP2018, a strain isolated from the oligotrophic part of the eastern tropical North Pacific (ETNP) oxygen minimum zone (OMZ), hosts the smallest of the fifteen genomes and yet the number of protein-coding genes it possesses is on par with the other strains. Its genome exhibits a distinct reduction in genes dedicated to Major Facilitator Superfamily transporters as well as biosynthetic enzymes. However, its core metabolic pathways are fully conserved. Our research indicates that the selective pressures of the ETNP OMZ favor a streamlined genome with reduced overall biosynthetic potential balanced by a stable set of core metabolisms and an expansion of mechanisms for nutrient acquisition. CONCLUSIONS: In summary, this study offers insights into the adaptation of fungi to the oligotrophic ocean and provides valuable information for understanding the ecological roles of fungi in the ocean.