Biogenic synthesis of multifunctional silver nanoparticles from <Emphasis Type="Italic">Rhodotorula glutinis</Emphasis> and <Emphasis Type="Italic">Rhodotorula mucilaginosa</Emphasis>: antifungal,catalytic and cytotoxicity activities

Silver nanoparticles (AgNPs) have several technological applications and may be synthetized by chemical, physical and biological methods. Biosynthesis using fungi has a wide enzymatic range and it is easy to handle. However, there are few reports of yeasts with biosynthetic ability to produce stable AgNPs. The purpose of this study was to isolate and identify soil yeasts (Rhodotorula glutinis and Rhodotorula mucilaginosa). After this step, the yeasts were used to obtain AgNPs with catalytic and antifungal activity evaluation. Silver Nanoparticles were characterized by UV–Vis, DLS, FTIR, XRD, EDX, SEM, TEM and AFM. The AgNPs produced by R. glutinis and R. mucilaginosa have 15.45?±?7.94 nm and 13.70?±?8.21 nm (average?±?SD), respectively, when analyzed by TEM. AgNPs showed high catalytic capacity in the degradation of 4-nitrophenol and methylene blue. In addition, AgNPs showed high antifungal activity against Candida parapsilosis and increase the activity of fluconazole (42.2% for R. glutinis and 29.7% for R. mucilaginosa), while the cytotoxicity of AgNPs was only observed at high concentrations. Finally, two yeasts with the ability to produce AgNPs were described and these particles showed multifunctionality and can represent a technological alternative in many different areas with potential applications.     

Lipid accumulation inRhodotorula glutinis on sugar cane molasses in single-stage continuous culture

Microbial lipids produced byRhodotorula glutinis grown in continuous culture with molasses under nitrogen-limiting conditions were evaluated and the effects of growth rate on fatty acid composition were studied. As the growth rate decreased, cell biomass, lipid content and lipid yield gradually increased. The maximum lipid content recorded was 39% (w/w) of dry cell biomass at a dilution rate of 0.04 h^–1. The growth rate also affected fatty acid composition: oleic acid decreased with decreasing growth rate while stearic acid increased.     

Effect of ammonium-N on malic enzyme and lipid production in Rhodotorula glutinis grown on monosodium glutamate wastewater

Rhodotorula glutinis, an oil producing strain, can utilize monosodium glutamate (MSG) wastewater as a raw material for lipid production. The effects of ammonium-N in the MSG wastewater (ammonium 15,000–25,000?mg/L, COD 30,000–50,000?mg/L) on cell growth, lipid accumulation and malic enzyme activity of R. glutinis have been studied. Four initial ammonium sulfate concentrations in the medium were set, which were 20, 60, 100, and 140?g/L. With an increase in the ammonium sulfate concentration, the uptake of ammonia nitrogen and lipid accumulation increased while the biomass decreased at 72?h. The maximum value of ammonia nitrogen consumption reached 5.77?g/L for an initial ammonium sulfate concentration of 140?g/L at 72?h. In addition, 60?g/L ammonium sulfate concentration may be an appropriate concentration for R. glutinis cultivation. The activity of the malic enzyme was measured and the results showed that there was a linear relationship between the intracellular lipid content and the total malic enzyme activity.     

Efficient concomitant production of lipids and carotenoids by oleaginous red yeast <Emphasis Type="Italic">Rhodotorula glutinis</Emphasis> cultured in palm oil mill effluent and application of lipids for biodiesel production

Rhodotorula glutinis TISTR 5159 is oleaginous red yeast that accumulates both lipids and carotenoids. It was cultured in palm oil mill effluent (POME) with only the addition of ammonium sulfate and Tween 20 as a suitable nitrogen source and surfactant, respectively. Response surface methodology (RSM) was applied to optimize initial chemical oxygen demand (COD) in POME, C/N ratio, and Tween 20 concentration for concomitant production of lipids and carotenoids. Among three investigated factors, C/N ratio contributed a significant effect upon lipid and carotenoids production. Analysis of response surface plots revealed that the optimum C/N ratio for the biomass was 140, while that for lipid content and carotenoids were higher at 180 and 170, respectively. The high level of the nitrogen source (with a low C/N ratio) enhanced the biomass, making the accumulation of lipids and carotenoids less preferable. Hence, the two-stage process was attempted as an optimal way for cell growth in the first stage and product accumulation in the second stage. The lipid yield and carotenoid production obtained in the two-stage process were higher than those in the one-stage process. In the semi-continuous fermentation, R. glutinis TISTR 5159 accumulated high lipid content and produced a considerably high concentration of carotenoids during long-term cultivation. Additionally, efficient COD removal by R. glutinis TISTR 5159 was observed. The biodiesel produced from yeast lipids was composed mainly of oleic and palmitic acids, similar to those from plant oil.     

Nickel(II) biosorption by <Emphasis Type="Italic">Rhodotorula glutinis</Emphasis>

The present study reports the feasibility of using Rhodotorula glutinis biomass as an alternative low-cost biosorbent to remove Ni(II) ions from aqueous solutions. Acetone-pretreated R. glutinis cells showed higher Ni(II) biosorption capacity than untreated cells at pH values ranging from 3 to 7.5, with an optimum pH of 7.5. The effects of other relevant environmental parameters, such as initial Ni(II) concentration, shaking contact time and temperature, on Ni(II) biosorption onto acetone-pretreated R. glutinis were evaluated. Significant enhancement of Ni(II) biosorption capacity was observed by increasing initial metal concentration and temperature. Kinetic studies showed that the kinetic data were best described by a pseudo-second-order kinetic model. Among the two-, three-, and four-parameter isotherm models tested, the Fritz-Schluender model exhibited the best fit to experimental data. Thermodynamic parameters (activation energy, and changes in activation enthalpy, activation entropy, and free energy of activation) revealed that the biosorption of Ni(II) ions onto acetone-pretreated R. glutinis biomass is an endothermic and non-spontaneous process, involving chemical sorption with weak interactions between the biosorbent and Ni(II) ions. The high sorption capacity (44.45 mg g⁻¹ at 25°C, and 63.53 mg g⁻¹ at 70°C) exhibited by acetone-pretreated R. glutinis biomass places this biosorbent among the best adsorbents currently available for removal of Ni(II) ions from aqueous effluents.     

Enthalpy content as a function of lipid accumulation in Rhodotorula glutinis

Microcalorimetry has been demonstrated to be a suitable on-line method for monitoring the lipid production phase of oleaginous yeasts. The choice of lipid extraction method for the oil accumulated by oleaginous yeasts is highly important both for accuracy when quantifying the lipid level and determining the fatty acid composition. The energy content of Rhodotorula glutinis increased from 23.0 kJ/g to 30.6 kJ/g dry biomass during the lipid-accumulating phase and was directly correlated to the analysed level of lipids, when an alkaline hydrolysis extraction method was used. Consequently, bomb-calorimetric measurements of the energy content were shown to be an indirect method of quantifying the lipid content in oleaginous yeasts. The fatty acid composition remained rather constant during the batch growth of Rh. glutinis with approximately 70% unsaturated C18 fatty acids. The high energy content as well as the fatty acid composition of Rh. glutinis makes this yeast a better candidate for use as aquaculture feed compared with the commonly used Saccharomyces cerevisiae.     

Single cell protein production from mandarin orange peel

Summary As the hydrolysis of mandarin orange peel with macerating enzyme (40°C, 24 h) produced 0.59 g g^–1 reducing sugar per dry peel compared to 0.36 by acid-hydrolysis (15 min at 120°C with 0.8 N H₂SO₄), the production of single cell protein (SCP) from orange peel was studied mostly using enzymatically hydrolyzed orange peel.When the enzymatically hydrolyzed peel media were used, the utilization efficiency of reducing sugars (%) and the growth yield from reducing sugars (g g^–1) were: 63 and 0.51 for Saccharomyces cerevisiae; 56 and 0.48 for Candida utilis; 74 and 0.69 for Debaryomyces hansenii and 64 and 0.70 for Rhodotorula glutinis. SCP production from orange peel by D. hansenii and R. glutinis were further studied. Batch cultures for 24 h at 30°C using 100 g dried orange peel produced 45 g of dried cultivated peel (protein content, 33%) with D. hansenii and 34 g (protein content, 50%) with R. glutinis, and 38 g (protein content, 44%) with a mixture of both yeasts.     

Biochemical characterization and evaluation of invertases produced from Saccharomyces cerevisiae CAT-1 and Rhodotorula mucilaginosa for the production of fructooligosaccharides

Abstract

Invertases are used for several purposes; one among these is the production of fructooligosaccharides. The aim of this study was to biochemically characterize invertase from industrial Saccharomyces cerevisiae CAT-1 and Rhodotorula mucilaginosa isolated from Cerrado soil. The optimum pH and temperature were 4.0 and 70?°C for Rhodotorula mucilaginosa invertase and 4.5 and 50?°C for Saccharomyces cerevisiae invertase. The pH and thermal stability from 3.0 to 10.5 and 75?°C for R. mucilaginosa invertase, respectively. The pH and thermal stability for S. cerevisiae CAT-1 invertase from 3.0 to 7.0, and 50?°C, respectively. Both enzymes showed good catalytic activity with 10% of ethanol in reaction mixture. The hydrolysis by invertases occurs predominantly when sucrose concentrations are ≤5%. On the other hand, the increase in the concentration of sucrose to levels above 10% results in the highest transferase activity, reaching about 13.3?g/L of nystose by S. cerevisiae invertase and 12.6?g/L by R. mucilaginosa invertase. The results demonstrate the high structural stability of the enzyme produced by R. mucilaginosa, which is an extremely interesting feature that would enable the application of this enzyme in industrial processes.     

Co‐hydrolysis of lignocellulosic biomass for microbial lipid accumulation

The herbaceous perennial energy crops miscanthus, giant reed, and switchgrass, along with the annual crop residue corn stover, were evaluated for their bioconversion potential. A co‐hydrolysis process, which applied dilute acid pretreatment, directly followed by enzymatic saccharification without detoxification and liquid–solid separation between these two steps was implemented to convert lignocellulose into monomeric sugars (glucose and xylose). A factorial experiment in a randomized block design was employed to optimize the co‐hydrolysis process. Under the optimal reaction conditions, corn stover exhibited the greatest total sugar yield (glucose + xylose) at 0.545 g g^?1 dry biomass at 83.3% of the theoretical yield, followed by switch grass (0.44 g g^?1 dry biomass, 65.8% of theoretical yield), giant reed (0.355 g g^?1 dry biomass, 64.7% of theoretical yield), and miscanthus (0.349 g g^?1 dry biomass, 58.1% of theoretical yield). The influence of combined severity factor on the susceptibility of pretreated substrates to enzymatic hydrolysis was clearly discernible, showing that co‐hydrolysis is a technically feasible approach to release sugars from lignocellulosic biomass. The oleaginous fungus Mortierella isabellina was selected and applied to the co‐hydrolysate mediums to accumulate fungal lipids due to its capability of utilizing both C5 and C6 sugars. Fungal cultivations grown on the co‐hydrolysates exhibited comparable cell mass and lipid production to the synthetic medium with pure glucose and xylose. These results elucidated that combining fungal fermentation and co‐hydrolysis to accumulate lipids could have the potential to enhance the utilization efficiency of lignocellulosic biomass for advanced biofuels production. Biotechnol. Bioeng. 2013; 110: 1039–1049. © 2012 Wiley Periodicals, Inc.     

Effect of various nutrient limitations on fatty acid production by Rhodotorula glutinis

The effects of nitrogen (N), phosphorus (P), zinc (Zn) or iron (Fe) limitations on lipid and especially -linolenic acid (ALA) synthesis in the oleaginous yeast Rhodotorula glutinis have been investigated. Exhaustion of the limiting element in the medium resulted in an enhancement of both the fatty acid cell content (FA/X) and the corresponding productivity. Except for Fe-limited media, lipid accumulation was always coupled with an increase in the yield of ALA synthesis, whereas in all the limiting conditions the productivity of ALA declined. Kinetic investigations showed that this decline resulted from a sharp decrease in the specific rates of ALA synthesis associated with slowing down of growth. A comparative study showed that the highest yields and specific rates of fatty acid and ALA synthesis were induced by P-limitation early in the lipid accumulation phase. However, above a FA/X of 15%, N-limiting conditions become more attractive for producing lipids. During P-limited growth a down-shift of temperature from 30° C to 25° C was shown ao reduce the range of FA/X values at which the specific rate of fatty acid synthesis was maximal but without any other effect on fatty acid formation. In contrast, the decreased temperature resulted in enhanced ALA production by maintaining the specific rate of synthesis near to the maximal value of 3.6 mg g _X ^–1 h^–1, where X^* is free fatty acid biomass, and increased the corresponding yield by a factor of up to three. Correspondence to: A. Pareilleux     

Caroteno-protein and exopolysaccharide production by co-cultures of Rhodotorula glutinis and Lactobacillus helveticus

The lactose-negative yeast Rhodotorula glutinis 22P and the homofermentative lactic acid bacterium Lactobacillus helveticus 12A were cultured together in a cheese whey ultrafiltrate containing 42 g L⁻¹ lactose. The chemical composition of the caroteno-protein has been determined. The carotenoid and protein contents are 248  μ g g⁻¹ dry cells and 48.2% dry weight. Carotenoids produced by Rhodotorula glutinis 22P have been identified as β-carotene 15%, torulene 10%, and torularhodin 69%. After separating the cell mass from the microbial association, the exopolysaccharides synthesized by Rhodotorula glutinis 22P were isolated from the supernatant medium in a yield of 9.2 g L⁻¹. The monosaccharide composition of the synthesized biopolymer was predominantly D-mannose (57.5%). Received 08 July 1996/ Accepted in revised form 11 December 1996     

Biotechnological conversion of waste cooking olive oil into lipid-rich biomass using Aspergillus and Penicillium strains

Aims: In this study, we have investigated the biochemical behaviour of Aspergillus sp. (five strains) and Penicillium expansum (one strain) fungi cultivated on waste cooking olive oil. The production of lipid‐rich biomass was the main target of the work. In parallel, the biosynthesis of other extracellular metabolites (organic acids) and enzyme (lipase) and the substrate fatty acid specificity of the strains were studied. Methods and Results: Carbon‐limited cultures were performed on waste oil, added in the growth medium at 15 g l^?1, and high biomass quantities were produced (up to c. 18 g l^?1, conversion yield of c. 1·0 g of dry biomass formed per g of fat consumed or higher). Cellular lipids were accumulated in notable quantities in almost all cultures. Aspergillus sp. ATHUM 3482 accumulated lipid up to 64·0% (w/w) in dry fungal mass. In parallel, extracellular lipase activity was quantified, and it was revealed to be strain and fermentation time dependent, with a maximum quantity of 645 U ml^?1 being obtained by Aspergillus niger NRRL 363. Storage lipid content significantly decreased at the stationary growth phase. Some differences in the fatty acid composition of both cellular and residual lipids when compared with the initial substrate fat used were observed; in various cases, cellular lipids more saturated and enriched with arachidic acid were produced. Aspergillus strains produced oxalic acid up to 5·0 g l^?1. Conclusions: Aspergillus and Penicillium strains are able to convert waste cooking olive oil into high‐added‐value products. Significance and Impact of the Study: Increasing fatty wastes amounts are annually produced. The current study provided an alternative way of biovalourization of these materials, by using them as substrates, to produce added‐value compounds.     

Biomass and carotenoid pigment production by patagonian native yeasts

Summary New yeast isolates from unexplored Patagonian habitats were studied for the production of biomass and carotenoids as the first step towards the selection of hyper-producing strains and the design of a process optimization approach. Patagonian yeast isolates considered as potential biomass and carotenoid sources were studied using ammonium sulphate and urea as nitrogen sources in semi-synthetic medium (MMS), and agro-industrial byproducts (cane molasses, corn syrup, raw malt extract) as carbon sources. Maximum pigment production (300 μg g⁻¹) was achieved by Rhodotorula mucilaginosa CRUB 0195 and by novel species Cryptococcus sp. CRUB 1046. β-carotene, torulene and torularhodin were the major carotenoids found.     

Accumulation of lipid byRhodotorula glutinis in continuous culture

Summary Rhodotorula glutinis accumulated 35% (w/w) lipid when grown nitrogen-limited in a chemostat at a dilution rate (D) of 0.02^–1 . At D = 0.10 h^–1, the lipid content was only 15% (w/w). Dual limitation of nitrogen and phosphate increased neither the amount of lipid produced nor the lipid yield (14g lipid per 100g glucose consumed). The fatty acid composition was unchanged by the growth rate.     

Spontaneous and radical-induced plasma membrane lipid peroxidation in differently oxidant-sensitive yeast species and its suppression by antioxidants

Formation of thiobarbituric acid-reactive substances (TBRS; nmol/mg lipids) indicative of lipid peroxidation was measured in whole cells and in isolated plasma membrane lipids from three yeast species differing in oxidant sensitivity (Schizosaccharomyces pombe, Saccharomyces cerevisiae andRhodotorula glutinis) after exposure to the Fenton reagent, Fe^II, H₂O₂,tert-butyl hydroperoxide (TBHP) and azo compounds (AAPH, ACHN). In whole cells, spontaneous TBRS formation rose in the sequenceS. pombe<S. cerevisiae<R. glutinis (1:∼5:∼7). Oxidants increased the TBRS production 13–18 fold in the sequence Fe^II∼TBHP>AAPH∼ACHN∼Fe-Fenton>H₂O₂. This increase need not be solely due to increased lipid peroxidation. In isolated plasma membrane lipids from all three species, the spontaneous TBRS production referred to 1 mg lipids was 9–13-fold higher than in whole cells. InS. pombe lipids, only TBHP increased the TBRS production. In lipids fromS. cerevisiae andR. glutinis, all added oxidants increased the spontaneous TBRS production 2–3 times in the sequence TBHP>ACHN>AAPH>Fe^II>Fe-Fenton>H₂O₂. Oxidant-induced TBRS production in both whole cells and isolated membrane lipids was partially suppressed by the lipid peroxidation inhibitors 2,6-di-tert-butyl-4-methylphenol (“butylated hydroxytoluene”; BHT) and the newly synthesized PYA12 compound. Both agents were more effective in isolated lipids than in whole cells and against OH-producing than against ROO-or RO-producing oxidants. Yeast membrane lipids, which are generally poor in polyunsaturated fatty acids, are thus subject to perceptible lipid peroxidation.     

Enhanced β-fructofuranosidase biosynthesis by Rhodotorula glutinis using Taguchi robust design method

The aim of this study was to produce β-fructofuranosidase enzyme by Rhodotorula glutinis SO28, using sugar beet (Beta vulgaris) as carbon source due to its high sucrose content and easy availability. β-Fructofuranosidase production was carried out in submerged fermentation. Taguchi orthogonal array (OA) design of experiment (DOE) method was employed for optimization process of β-fructofuranosidase production by R. glutinis SO28. An OA layout of L16 was constructed with five influential factors on β-fructofuranosidase biosynthesis namely, carbon source (sugar beet), initial pH, incubation temperature, agitation speed and incubation time. The average results of β-fructofuranosidase yield obtained from the determined 16 batches were processed with Minitab^® 16.2.3 software at “larger is better” as quality character. The results showed that the maximum β-fructofuranosidase activity was obtained as 21.11?±?0.47?U/mL, which was close to the predicted result (21.78?±?0.43?U/mL). Consequently, sugar beet can be suggested as an economical substrate for β-fructofuranosidase production. Besides, use of Taguchi DOE enhanced enzyme activity about 3-fold when compared with unoptimized condition.     

Direct bioconversion of rice residue from canteen waste into lipids by new amylolytic oleaginous yeast Sporidiobolus pararoseus KX709872

The new amylolytic oleaginous red yeast, Sporidiobolus pararoseus KX709872, produced both α-amylase (540?±?0.09?mU/mL) and amyloglucosidase (23?±?0.00?mU/mL) and showed good ability to directly convert rice residue from canteen waste to biomass and lipids. Effects of medium composition and cultivation conditions on growth and lipid accumulation for strain KX709872 were investigated under shaking flask and upscaling levels. At C?:?N ratio of 25?:?1, pH 5.45, 22.36°C, and 199.40?rpm for 7 days, volumetric production of biomass and lipids, lipid content, and lipid productivity reached 17.69?±?0.44, 8.35?±?0.19?g/L, 49.48?±?0.41% (w/w), and 1.67?±?0.11?g/L/day, respectively. Production of lipids was also implemented in 5.0-L stirred tank bioreactor with 2.5?L of optimized medium at 300?rpm and 3.0 vvm for 5 days. Volumetric production of biomass and lipids, lipid content, and lipid productivity were 16.33?±?0.49, 8.75?±?0.13?g/L, 56.61?±?0.04% (w/w), and 2.19?±?0.03?g/L/day, respectively. Meanwhile, the fatty acids of lipids from strain KX709872 had high oleic acid content (60?62%) which was similar to those of vegetable oils, indicating that these lipids are promising as an alternative biodiesel feedstock. Moreover, the biodiesel derived from lipids of strain KX709872 had properties satisfying the criteria of ASTM D6751 and EN 14214 standards.     

Lead uptake and potentiometric titration studies with live and dried cells of Rhodotorula glutinis

The ability of live cells (LC), freeze dried cells (FDC) and oven dried cells (ODC) of the yeast Rhodotorula glutinis to remove lead from aqueous solution has been studied. Discernible differences were found between the biosorption properties of LC and the other two types of cell preparation. The LC preparation exhibited an uptake level of about 12 mg g⁻¹ in a batch contactor with a biomass dosage of 2 g l⁻¹ and an initial lead concentration of 100 mg l⁻¹. This compared with, respectively, about 26 and 30 mg g⁻¹ for the FDC and ODC biosorbents under the same experimental conditions. It is seen that the level of lead uptake by the two latter biosorbents was increased to, respectively, 2.2- and 2.5-fold of the level observed for the LC preparation. The superior performance of the FDC and ODC biosorbents in the lead binding process was attributed to the presence of additional binding sites on their cell wall surfaces as indicated by potentiometric titration data. These binding sites were ascribed to carboxylic and phosphoric groups, which are the primary sites of divalent metal complexation. Modeling of the titration data revealed that subjecting R. glutinis biomass to freeze drying or oven drying increased its proton binding site concentration by a factor of 3. It appears that the two simple physical treatments were able to compromise the R. glutinis cell wall structure in such a way as to make sites normally inaccessible to become active in proton and lead binding.     

Outdoor pilot-scale production of Botryococcus braunii in panel reactors

In this paper, the outdoor production of Botryococcus braunii in pilot-scale panel reactors (0.4?m³) is studied under uncontrolled conditions at a location close to the Atacama Desert (Chile). Discontinuous experiments were performed on different dates to determine the feasibility of the culture and the influence of environmental conditions on the system yield. Data showed that solar radiation is a major parameter in determining system yield, the average irradiance inside the culture determining both the growth rate and biomass productivity. A maximum specific growth rate of 0.09?day^?1 and biomass productivity of 0.02?g?L^?1?day^?1 (dry weight) were measured in discontinuous mode, at an average irradiance of 60?μE?m^?2?s^?1. With respect to lipids, a productivity of 2.5?mg?L^?1?day^?1 was obtained under favourable growth conditions; no accumulation of lipids at the stationary phase was observed. To confirm this behaviour, a semicontinuous culture was performed at 0.04?day^?1 in a larger reactor (1?m³). In this experiment, the biomass concentration and productivity was 0.3?g?L^?1 and 0.015?g?L^?1?day^?1, respectively. The lipid content and productivity was 15.6% and 2.4?mg?L^?1?day^?1, respectively, the mean average irradiance inside the reactor being 60?μmol photons?m^?2?s^?1. The light path of the reactor determines the light availability, thus determining also the biomass concentration and productivity of the reactor once the dilution rate is fixed. Experimentally, biomass productivity of 0.015?g?L^?1?day^?1 was determined for a light path of 0.15?m, but this can be increased by more than three times for a light path of 0.1?m. These data confirm that this alga can be produced outdoors in a secure form, the culture yield improving when optimal conditions are applied, the data reported here establishing the starting point for the development of the process.     

Overexpression of malic enzyme (ME) of Mucor circinelloides improved lipid accumulation in engineered Rhodotorula glutinis

The oleaginous yeast Rhodotorula glutinis has been known to be a potential feedstock for lipid production. In the present study, we investigated the enhancement of expression of malic enzyme (ME; NADP⁺ dependent; EC 1.1.1.40) from Mucor circinelloides as a strategy to improve lipid content inside the yeast cells. The 26S rDNA and 5.8S rDNA gene fragments isolated from Rhodotorula glutinis were used for homologous integration of ME gene into R. glutinis chromosome under the control of the constitutively highly expressed gene phosphoglycerate kinase 1 to achieve stable expression. We demonstrated that by increasing the expression of the foreign ME gene in R. glutinis, we successfully improved the lipid content by more than twofold. At the end of lipid accumulation phrase (96 h) in the transformants, activity of ME was increased by twofold and lipid content of the yeast cells was increased from 18.74 % of the biomass to 39.35 %. Simultaneously, there were no significant differences in fatty acid profiles between the wild-type strain and the recombinant strain. Over 94 % of total fatty acids were C_16:0, C_18:0, C_16:1, C_18:1, and C_18:2. Our results indicated that heterologous expression of NADP⁺-dependent ME involved in fatty acid biosynthesis indeed increased the lipid accumulation in the oleaginous yeast R. glutinis.