torulene and torularhodin

Torulene and torularhodin represent the group of carotenoids and are synthesized by yeasts and fungi. The most important producers of these two compounds include yeasts of Rhodotorula and Sporobolomyces genera. The first reports confirming the presence of torulene and torularhodin in the cells of microorganisms date to the 1930s and 1940s; however, only in the past few years, the number of works describing the properties of these compounds increased. These compounds have strong anti-oxidative and anti-microbial properties, and thus may be successfully used as food, feedstock, and cosmetics additives. In addition, tests performed on rats and mice showed that both torulene and torularhodin have anti-cancerous properties. In order to commercialize the production of these two carotenoids, it is necessary to obtain highly efficient yeast strains, for example, via mutagenization and optimization of cultivation conditions. Further studies on the activity of torulene and torularhodin on the human body are also needed.

Topics: Animals; Carotenoids; Humans; Rats; Rats, Wistar

Rhodotorula glutinis is capable of synthesizing numerous valuable compounds with a wide industrial usage. Biomass of this yeast constitutes sources of microbiological oils, and the whole pool of fatty acids is dominated by oleic, linoleic, and palmitic acid. Due to its composition, the lipids may be useful as a source for the production of the so-called third-generation biodiesel. These yeasts are also capable of synthesizing carotenoids such as β-carotene, torulene, and torularhodin. Due to their health-promoting characteristics, carotenoids are commonly used in the cosmetic, pharmaceutical, and food industries. They are also used as additives in fodders for livestock, fish, and crustaceans. A significant characteristic of R. glutinis is its capability to produce numerous enzymes, in particular, phenylalanine ammonia lyase (PAL). This enzyme is used in the food industry in the production of L-phenylalanine that constitutes the substrate for the synthesis of aspartame-a sweetener commonly used in the food industry.

Topics: beta Carotene; Biofuels; Biomass; Carotenoids; Enzymes; Fatty Acids; Industrial Microbiology; Linoleic Acid; Oleic Acid; Palmitic Acid; Phenylalanine; Phenylalanine Ammonia-Lyase; Rhodotorula

Olive mill wastewater (OMW) is a zero-cost substrate for numerous value-added compounds. Although several studies on the production of lipids and carotenoids by Rhodotorula glutinis in OMW exist, none of them has specifically focused on the conditions for a target lipid or carotenoid. This study presents cultivation conditions that selectively stimulate the cell biomass, individual carotenoids and lipids. It was found that supplemental carbon and nitrogen sources as well as illumination affected cell biomass the most. High temperature, low initial pH, illumination, lack of urea and presence of glycerol stimulated the lipid synthesis. The highest total lipid content obtained in undiluted OMW supplemented with urea was 11.08 ± 0.17% (w/w) whilst it was 41.40 ± 0.21% (w/w) when supplemented with glycerol. Moreover, the main fatty acid produced by R. glutinis in all media was oleic acid, whose fraction reached 63.94 ± 0.58%. Total carotenoid yield was significantly increased with low initial pH, high temperature, illumination, certain amounts of urea, glycerol and cultivation time. Up to 192.09 ± 0.16 μg/g cell carotenoid yield was achieved. Torularhodin could be selectively produced at high pH, low temperature and with urea and glycerol supplementation. To selectively induce torulene synthesis, cultivation conditions should have low pH, high temperature and illumination. In addition, low pH, high temperature and urea supplementation served high production of β-carotene. Up to 85.40 ± 0.76, 80.67 ± 1.40 and 39.45 ± 0.69% of torulene, torularhodin and β-carotene, respectively, were obtained under selected conditions. KEY POINTS: • Cultivation conditions selectively induced target carotenoids and lipids • 41.40 ± 0.21% (w/w) lipid content and 192.09 ± 0.16 μg/g cell carotenoid yield were achieved • Markedly high selectivity values for torularhodin and torulene were achieved.

Topics: beta Carotene; Carotenoids; Fatty Acids; Glycerol; Olea; Rhodotorula; Wastewater

Carotenoids represent a diverse class of aliphatic C40 molecules with a variety of applications in the food and pharmaceutical industries. Sporidiobolus pararoseus NGR produces various carotenoids, including torulene, torularhodin and β-carotene. Salt stress significantly increases the torulene accumulation of S. pararoseus NGR. However, little is known, about the molecular mechanisms underlying the increased torulene biosynthesis. In this work, we investigated the effects of NaCl treatment on the contents of carotenoids (both qualitatively and quantitatively) and transcriptome. A total of 12.3 Gb of clean bases were generated in six cDNA libraries. These bases were de novo assembled into 9,533 unigenes with an average length of 1,654 nt and N50 of 2,371 nt. Transcriptome analysis revealed that of 3,849 differential expressed genes (DEGs) in response to salt stress, 2,019 were up-regulated, and 1,830 were down-regulated. Among these DEGs, we identified three carotenogenic genes crtE, crtYB, and crtI. In addition, fourteen candidate genes were predicted to participate in the conversion from torulene to torularhodin. Our findings should provide insights into the mechanisms of carotenoid biosynthesis and salt-tolerance of S. pararoseus NGR.

Topics: Basidiomycota; beta Carotene; Biosynthetic Pathways; Carotenoids; Fungal Proteins; Gene Expression Profiling; Gene Expression Regulation, Bacterial; Salt Stress; Sodium Chloride

Topics: Apoptosis; bcl-2-Associated X Protein; Blotting, Western; Carotenoids; Cells, Cultured; Gene Expression Regulation; Humans; Hydrogen Peroxide; Male; Oxidants; Oxidative Stress; Prostate; Proto-Oncogene Proteins c-bcl-2; Real-Time Polymerase Chain Reaction; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Stromal Cells

The red yeast Sporobolomyces ruberrimus H110 was able to use glycerol as a carbon source. The highest concentration (0.51gL(-1)) and productivity (0.0064gL(-1)h(-1)) of carotenoids were achieved when raw glycerol from biodiesel production, containing around 1gL(-1) of fatty acids, was used as the carbon source, which represented increases of 27% and 1.5×, respectively, in relation to pure glycerol. Mass spectrometry analysis led to the identification of four carotenoids in the fermented samples, torularhodin, torulene, β-carotene and γ-carotene. The use of raw glycerol also enhanced the proportion of torularhodin (69% against 59% in pure glycerol). The addition of individual fatty acids (palmitic, stearic, oleic and linoleic acids) to pure glycerol resulted in increases between 15% and 25% in maximum concentration and between 1.6× and 2.0× in productivity of carotenoids. The presence of palmitic and oleic acids increased the torularhodin proportion to 66%.

Topics: Basidiomycota; beta Carotene; Biofuels; Bioreactors; Carotenoids; Chromatography, Gas; Chromatography, High Pressure Liquid; Fatty Acids; Fermentation; Glycerol; Linoleic Acids; Mass Spectrometry; Oleic Acids; Palmitic Acids; Stearic Acids

Torulene and torularhodin are two of the principal carotenoids in Sporidiobolus pararoseus and have a similar structure to that of lycopene. The present study was to elucidate the anti-cancer activity of torulene and torularhodin in vivo with lycopene as a control. Nude mice were orally supplemented every day with a low or high dose [9 or 18 mg/kg body weight (BW)] of lycopene, torularhodin or torulene. Two weeks after the supplementation, mice were injected once with hormone-independent prostatic carcinoma PC-3 cells. When the tumor of the control group load exceeded 200 mm(3), mice were killed and the study was terminated. Compared with the controls, high-carotenoid supplementation lowered the mean number of tumors from 248.13 ± 28.74 to 50.83 ± 7.63, 70.34 ± 6.77, and 60.53 ± 6.78 mm(3) (P < 0.05, n = 8) by, respectively. Histological examination showed tumor degeneration, apoptosis and necrosis presented at the end of the experiment. Quantitative polymerase chain reaction and immunohistochemistry results showed Bcl-2 expression of the control group was higher than that of the carotenoid-treated group while the expression of Bax was lower than the carotenoid-treated group. High-carotenoid supplementation also increased the mRNA expressions of caspase-3, 8 and 9 in tumor tissues. These results show that both torulene and torularhodin supplementation inhibit the growth of prostate cancer in nude mice and suggest that such an action is associated the apoptosis of tumor cells.

Topics: Administration, Oral; Animals; Antineoplastic Combined Chemotherapy Protocols; Carotenoids; Cell Line, Tumor; Cell Proliferation; Dose-Response Relationship, Drug; Humans; Male; Mice; Mice, Inbred BALB C; Mice, Nude; Prostatic Neoplasms

The pigments from Rhodotorula glutinis were separated by using thin layer chromatography, and the result showed that Rhodotorula glutinis cells could synthesize at least three kinds of pigments, which were beta-carotene, torulene, and torularhodin. The Raman spectra based on the three pigments were acquired, and original spectra were preprocessed by background elimination, baseline correction, and three-point-smoothing, then the averaged spectra from different pigments were investigated, and the result indicated that Raman shift which represents C-C bond was different, and the wave number of beta-carotene demonstrated the largest deviation, finally torulene and torularhodin in Rhodotorula glutinis had more content than beta-carotene. Quantitative analysis of Raman peak height ratio revealed that peak height ratio of pigments showed little difference, which could be used as parameters for further research on living cells, providing reference content of pigments. The above results suggest that Raman spectroscopy combined with thin layer chromatography can be applied to analyze pigments from Rhodotorula glutinis, provides abundant information about pigments, and serves as an effective method to study pigments.

Topics: beta Carotene; Carotenoids; Chromatography, Thin Layer; Rhodotorula; Spectrum Analysis, Raman

Yeasts of the genera Rhodotorula are able to synthesize different pigments of high economic value like β-carotene, torulene, and torularhodin, and therefore represent a biotechnologically interesting group of yeasts. However, the low production rate of pigment in these microorganisms limits its industrial application. Here we describe some strategies to obtain hyperpigmented mutants of Rhodotorula mucilaginosa by means of ultraviolet-B radiation, the procedures for total carotenoids extraction and quantification, and a method for identification of each pigment.

Topics: beta Carotene; Carotenoids; Chromatography, High Pressure Liquid; Metabolic Engineering; Mutagenesis; Mutation; Rhodotorula

A simple method for the extraction of carotenoid pigments from frozen wet cells of red yeasts (Basidiomycota) and their analysis by reversed-phase HPLC using a C(18) column and a water/acetone solvent system is described. Typical red yeast carotenoids belonging to an oxidative series from the monocyclic gamma-carotene to 2-hydroxytorularhodin and from the bicyclic beta-carotene to astaxanthin were separated. Pigment identity was confirmed by LC-atmospheric pressure chemical ionisation (APCI) mass spectrometry using similar chromatographic conditions.

Topics: Basidiomycota; beta Carotene; Carotenoids; Chromatography, High Pressure Liquid; Molecular Structure; Reproducibility of Results; Xanthophylls

Two red Cystofilobasidium spp. isolated from spring sap-flows of Betula pendula were analysed for their carotenoid content. In Cystofilobasidium infirmominiatum, three unusual pigments were detected and identified by structure elucidation as oxidised torulene derivatives. These included 16'-hydroxytorulene and torularhodinaldehyde, two carotenoids known so far only from chemical synthesis or as postulated biosynthetic intermediates en route to torularhodin. Unprecedented formation of beta-apo-2'-carotenal was also observed. The production of these pigments in pure culture was dependent on enhanced oxidative stress caused by cultivation in well-aerated (indented) flasks with or without 2% ethanol (16'-hydroxytorulene), or with 100 microM duroquinone (torularhodinaldehyde and beta-apo-2'-carotenal). Among these three pigments, only 16'-hydroxytorulene was detected in C. capitatum. Torularhodin, a common end product of carotenoid oxidation in red yeasts, was not produced by either species under any incubation conditions. Biosynthetic aspects of incomplete oxidation of torulene by these Cystofilobasidium spp. are discussed.

Topics: Basidiomycota; Carotenoids; Chromatography, High Pressure Liquid; Magnetic Resonance Spectroscopy; Molecular Structure; Oxidative Stress

Eighteen yeast species of the genera Rhodotorula, Rhodosporidium, Sporobolomyces, and Sporidiobolus, each one represented by its type strain, were investigated with the objective of evaluating their carotenoid composition. The pigments were extracted from yeast cells, quantified by high pressure liquid chromatography diode array detector and the main compounds were confirmed by atmospheric pressure chemical ionization quadrupole mass spectrometry. Significant (P < 0.01) differences among several species and (or) genera were observed. Thirteen strains were seen to be able to produce carotenoids, from 16.4 to 184 microg/g cell dry mass and from 6.0 to 1993.4 microg/L culture. The main carotenoids produced were identified as torularhodin, torulene, gamma-carotene, and beta-carotene. The correlation matrix calculated on the basis of the carotenoid composition data matrix indicated significant (P < 0.01) relationships between torulene and torularhodin (r = 0.81), gamma-carotene and torulene (r = 0.49), beta-carotene and torulene (r = -0.72), as well as beta-carotene and gamma-carotene (r = 0.64). These significant correlation coefficients may suggest that species belonging to the genera Rhodosporidium, Sporobolomyces, and Sporidiobolus possess a carotenoid biosynthetic pathway analogous to that elsewhere postulated for Rhodotorula species.

Topics: Basidiomycota; beta Carotene; Carotenoids; Chromatography, High Pressure Liquid; Mass Spectrometry; Rhodotorula; Yeasts

A carotenoid-producing yeast strain, isolated from the sub-arctic, marine copepod Calanus finmarchicus, was identified as Rhodosporidium babjevae (Golubev) according to morphological and biochemical characteristics and phylogenetic inference from the small-subunit ribosomal RNA gene sequence. The total carotenoids content varied with cultivation conditions in the range 66-117 microg per g dry weight. The carotenoid pool, here determined for the first time, was dominated by torularhodin and torulene, which collectively constituted 75-91% of total carotenoids under various regimes of growth. Beta-carotene varied in the range 5-23%. A high-peptone/low-yeast extract (weight ratio 38:1) marine growth medium favoured the production of torularhodin, the carotenoid at highest oxidation level, with an average of 63% of total carotenoids. In standard yeast medium (YM; ratio 1.7:1), torularhodin averaged 44%, with increased proportions of the carotenes, torulene and beta-carotene. The anticipated metabolic precursor gamma-carotene (beta,psi-carotene) constituted a minor fraction (

Topics: beta Carotene; Carotenoids; Chromatography, High Pressure Liquid; Molecular Structure; Phylogeny; Polymerase Chain Reaction; Rhodotorula

Carotenogenesis of the lactose-negative yeast Rhodotorula rubra GED5 was studied by cocultivation with Kluyveromyces lactis MP11 in whey ultrafiltrate (WU) (35, 50, and 70 g of lactose/L). Maximum yields of cell mass (24.3 g/L) and carotenoids (10.2 mg/L of culture fluid or 0.421 micro g/g of dry cells) were obtained by growing the microbial association in WU (50 g of lactose/L) in a fermentor with an airflow rate of 0.8 L/(L.min), agitation of 220 rpm, and temperature of 30 degrees C. The identified carotenoid pigments-beta-carotene, torulene, and torularhodin-reached maximum concentrations (133, 26.9, and 222.3 microg/g of dry cells, respectively) on d 5 for torulene and d 6 for beta-carotene and torularhodin.

Topics: Animals; beta Carotene; beta-Galactosidase; Biomass; Carotenoids; Coculture Techniques; Hydrogen-Ion Concentration; Industrial Microbiology; Kluyveromyces; Lactose; Milk; Rhodotorula; Ultrafiltration

Enhancement in the production of beta-carotene by the hyper producer mutant 32 of Rhodotorula glutinis by manipulation of temperature and illumination.. Growth and beta-carotene production was investigated in a 1 litre fermenter at different temperature and illumination conditions. The optimum temperature for growth and beta-carotene production was 30 and 20 degrees C, respectively. At 30 degrees C, beta-carotene production was 125 +/- 2 mg l-1 and accounted for 66% of the total carotenoids in 72 h; at 20 degrees C, it was 250 +/- 7 mg l-1 and accounted for 92% of total carotenoid content. Continuous illumination of the fermenter by 1000 lx white light hampered growth as well as carotenoid synthesis. At 30 degrees C, illuminating the fermenter in late logarithmic phase resulted in a 58% increase in beta-carotene production with a concurrent decrease in torulene; at 20 degrees C, however, it showed no appreciable increase.. Proper manipulation of culture conditions enhanced beta-carotene production by R. glutinis which makes it a significant source of beta-carotene.

Topics: beta Carotene; Carotenoids; Glucose; Light; Mutation; Rhodotorula; Temperature

Investigations on the production of red pigments by Rhodotorula glutinis on raw substrates of agro-industrial origin may be considered of interest because they represent the first approach to the utilization of these raw materials for biotechnological purposes.. Rhodotorula glutinis DBVPG 3853 was batch and fed-batch co-cultured with Debaryomyces castellii DBVPG 3503 in a medium containing corn syrup as the sole carbon source. Fed-batch co-cultures gave a volumetric production of 8.2 mg total carotenoid l(-1), about 150% of that observed in batch co-cultures. The different carotenoid pigments (beta-carotene, torulene, torularhodin) were quantified.. Oligosaccharides and dextrins of corn syrup could be used profitably for pigment production by R. glutinis DBVPG 3853-D. castellii DBVPG 3503 in co-culture.. The above results suggest that the red yeasts belonging to the genus Rhodotorula may have industrial relevance as carotenoid producers.

Topics: beta Carotene; Carotenoids; Chromatography, High Pressure Liquid; Coculture Techniques; Rhodotorula; Saccharomycetales; Zea mays

Several wild strains and mutants of Rhodotorula spp. were screened for growth, carotenoid production and the proportion of -carotene produced in sugarcane molasses. A better producer, Rhodotorula glutinis mutant 32, was optimized for carotenoid production with respect to total reducing sugar (TRS) concentration and pH. In shake flasks, when molasses was used as the sole nutrient medium with 40 g l(-1) TRS, at pH 6, the carotenoid yield was 14 mg l(-1) and -carotene accounted for 70% of the total carotenoids. In a 14-l stirred tank fermenter, a 20% increase in torulene content was observed in plain molasses medium. However, by addition of yeast extract, this effect was reversed and a 31% increase in -carotene content was observed. Dissolved oxygen (DO) stat fed-batch cultivation of mutant 32 in plain molasses medium yielded 71 and 185 mg l(-1) total carotenoids in double- and triple-strength medium, respectively. When supplemented with yeast extract, the yields were 97 and 183 mg l(-1) total carotenoid with a 30% increase in -carotene and a simultaneous 40% decrease in torulene proportion. Higher cell mass was also achieved by double- and triple-strength fed-batch fermentation.

Topics: beta Carotene; Biomass; Bioreactors; Carotenoids; Culture Media; Fermentation; Glucose; Hydrogen-Ion Concentration; Industrial Microbiology; Kinetics; Magnoliopsida; Molasses; Mutation; Nitrogen; Rhodotorula; Time Factors

Simpson, Kenneth L. (University of California, Davis), T. O. M. Nakayama, and C. O. Chichester. Biosynthesis of yeast carotenoids. J. Bacteriol. 88:1688-1694. 1964.-The biosynthesis of carotenoids was followed in Rhodotorula glutinis and in a new strain, 62-506. The treatment of the growing cultures by methylheptenone, or ionone, vapors permitted observations of the intermediates in the biosynthetic pathway. On the basis of concentration changes and accumulation in blocked pathways, the sequence of carotenoid formation is postulated as phytoene, phytofluene, zeta-carotene, neurosporene, beta-zeacarotene, gamma-carotene, torulin, a C(40) aldehyde, and torularhodin. Torulin and torularhodin were established as the main carotenoids of 62-506.

Topics: Aldehydes; Carotenoids; Chromatography; Hydrocarbons; Ketones; Metabolism; Pharmacology; Research; Saccharomyces cerevisiae; Xanthophylls; Yeasts