Docosahexaenoic Acid Yield Research Articles

Metabolomics analysis of phytohormone gibberellin improving lipid and DHA accumulation in Aurantiochytrium sp.

In this study, the phytohormone gibberellin was evaluated for its effect on lipid and docosahexaenoic acid (DHA) accumulation in Aurantiochytrium sp.YLH70. It was found that 4mg/mL of gibberellin increased biomass, lipid and DHA yield of Aurantiochytrium sp.YLH70 by 14.4%, 43.6% and 79.1%, respectively. Moreover, a GC–MS metabolomics method combined with a multivariate analysis was applied to reveal the metabolic mechanisms responsible for the increased lipid and DHA accumulation. The principal component analysis (PCA) revealed that metabolomics profiles from all groups were discriminated. 38 metabolites identified by the partial least-squares-discriminant analysis (PLS-DA) were responsible for responding to gibberellin treatment. Metabolic pathway analysis showed that gibberellin accelerated the rate of utilization of glucose, and metabolites in fatty acids biosynthesis and mevalonate pathway were increased, while metabolites in glycolysis and TCA cycle were decreased in Aurantiochytrium sp.YLH70. Moreover, the anti-stress mechanism in Aurantiochytrium sp.YLH70 might be induced by gibberellin.

Adaptive evolution of Schizochytrium sp. by continuous high oxygen stimulations to enhance docosahexaenoic acid synthesis

Adaptive laboratory evolution (ALE) is an effective method in changing the strain characteristics. Here, ALE with high oxygen as a selection pressure was applied to improve the production capacity of Schizochytrium sp. Results showed that cell dry weight (CDW) of endpoint strain was 32.4% higher than that of starting strain. But slight lipid accumulation impairment was observed. These major performance changes were accompanied with enhanced isocitrate dehydrogenase enzyme activity and reduced ATP:citrate lyase enzyme activity. And a serious decrease of 62.6% in SDHA 140rpm→170rpm was observed in the endpoint strain. To further study the docosahexaenoic acid (DHA) production ability of evolved strain, fed-batch strategy was applied and 84.34g/L of cell dry weight and 26.40g/L of DHA yield were observed. In addition, endpoint strain produced greatly less squalene than starting strain. This work demonstrated that ALE may be a promising tool in modifying microalga strains.

Effect of malate on docosahexaenoic acid production from Schizochytrium sp. B4D1

Background: Malate involves in the citrate/malate and transhydrogenase cycles to provide precursors for docosahexaenoic acid (DHA) synthesis. The optimal strategy was investigated for increasing DHA production in Schizochytrium species during fermentation. Results: DHA production increased by 47% and reached 5.51 g/L when 4 g malate/L was added during the rapid lipid accumulation stage in shake-flasks culture. Inducing effects of malate was further investigated through the analysis of three kinetic parameters, including specific cell growth rate (μ), specific glucose consumption rate (q Glu ) and DHA formation rate (q DHA ). DHA concentration was enhanced through a novel fed-batch strategy to a maximum value of 30.7 g/L, giving a yield of 0.103 g DHA/g glucose and a productivity of 284 mg L - 1 h - 1 . Conclusion: A novel malate feeding strategy was developed that enhanced DHA yield and productivity of Schizochytrium species which may offer a desirable method for industrial applications. Normal 0 21 false false false EN-US ZH-CN X-NONE /* Style Definitions */ table.MsoNormalTable {mso-style-name:Tabla normal; mso-tstyle-rowband-size:0; mso-tstyle-colband-size:0; mso-style-noshow:yes; mso-style-priority:99; mso-style-parent:; mso-padding-alt:0cm 5.4pt 0cm 5.4pt; mso-para-margin:0cm; mso-para-margin-bottom:.0001pt; mso-pagination:widow-orphan; font-size:10.5pt; mso-bidi-font-size:11.0pt; font-family:Calibri,sans-serif; mso-ascii-font-family:Calibri; mso-ascii-theme-font:minor-latin; mso-hansi-font-family:Calibri; mso-hansi-theme-font:minor-latin; mso-font-kerning:1.0pt; mso-ansi-language:EN-US; mso-fareast-language:ZH-CN;}

Improvement of Omega-3 Docosahexaenoic Acid Production by Marine Dinoflagellate Crypthecodinium cohnii Using Rapeseed Meal Hydrolysate and Waste Molasses as Feedstock.

Rapeseed meal and waste molasses are two important agro-industrial by-products which are produced in large quantities. In this study, solid state fermentation and fungal autolysis were performed to produce rapeseed meal hydrolysate (RMH) using fungal strains of Aspergillus oryzae, Penicillium oxalicum and Neurospora crassa. The hydrolysate was used as fermentation feedstock for heterotrophic growth of microalga Crypthecodinium cohnii that produce docosahexaenoic acid (DHA). The addition of waste molasses as a supplementary carbon source greatly increased the biomass and DHA yield. In the batch fermentations using media composed of diluted RMH (7%) and 1-9% waste molasses, the highest biomass concentration and DHA yield reached 3.43 g/L and 8.72 mg/L, respectively. The algal biomass produced from RMH and molasses medium also had a high percentage of DHA (22-34%) in total fatty acids similar to that of commercial algal biomass. RMH was shown to be rich in nitrogen supply comparable to the commercial nitrogen feedstock like yeast extract. Using RMH as sole nitrogen source, waste molasses excelled other carbon sources and produced the highest concentration of biomass. This study suggests that DHA production of the marine dinoflagellate C. cohnii could be greatly improved by concomitantly using the cheap by-products rapeseed meal hydrolysate and molasses as alternative feedstock.

Improved synthesis of docosahexaenoic acid (DHA) using Schizochytrium limacinum SR21 and sustainable media

Docosahexaenoic acid (DHA, C22:6) is an omega-3 fatty acid having beneficial health effects in human beings, though its production is severely hampered due to higher costs and lower yields from the synthesis. The current work concentrates on the intensification in the biomass production and DHA yield using Schizochytrium limacinum SR21. The effect of different media supplements on the DHA yields has been investigated. Citric acid which is a major intermediate in TCA (tricarboxylic acid) cycle proved better among different organic acids used in the work in terms of the yield of both biomass and DHA. The role of arginine and hydrophobic amino acids like tryptophan and tyrosine for the enhanced DHA production has also been illustrated. Monosaccharides (glucose and fructose) and glycerol also resulted in good cell growth and DHA yield. Maximum productivity of 5.7g/l DHA has been obtained when glucose and glycerol were used simultaneously as carbon source. It has also been established that different steep liquors, tomato juice and sweet water can be used as possible cheaper substrates for the production of DHA from S. limacinum SR21.

Production of High Docosahexaenoic Acid by <i>Schizochytrium</i> sp. Using Low-cost Raw Materials from Food Industry

The low-cost substrates from food industry, including maize starch hydrolysate and soybean meal hydrolysate, were used to produce docosahexaenoic acid (DHA) by Schizochytrium limacinum OUC88. Glucose derived from maize starch hydrolysate was used as the carbon source and soybean meal hydrolysate as the nitrogen sources. In 10L bioreactor fermentation, by using the soybean meal hydrolysate as the main nitrogen source, the biomass of Schizochytrium limacinum OUC88 reached 85.27 g L(-1), and the yields of DHA was 20.7g L(-1). As a comparison, when yeast extract was used as the main nitrogen source, the yields of biomass and DHA were 68.93 g L(-1) and 13.3 g L(-1), respectively. From the results of this study, these hydrolysates can provide all the nutrients required for high-density cultivation of S. limacinum OUC88 and DHA production, that will improve the economical and competitive efficiency of commercial DHA production.

A strategy for the highly efficient production of docosahexaenoic acid by Aurantiochytrium limacinum SR21 using glucose and glycerol as the mixed carbon sources

Glucose and glycerol are useful carbon sources for the cultivation of Aurantiochytrium limacinum SR21. Glucose facilitates rapid growth and lipid synthesis, and glycerol promotes the accumulation of docosahexaenoic acid (DHA) in A. limacinum SR21. To improve the DHA productivity of A. limacinum SR21, shake flask and fed-batch cultures were performed using glucose and glycerol as mixed carbon sources (MCSs). Along with optimization of the MCSs, the best DHA yield and productivity (32.36g/L and 337.1mg/L/h) were obtained via fed-batch fermentation with maintenance of a constant air supply. The DHA productivity was 15.24% higher than that obtained using glucose as single carbon source (SCS). This study presents a highly efficient and economic strategy for the production of DHA by A. limacinum SR21.

Efficient production of triacylglycerols rich in docosahexaenoic acid (DHA) by osmo-heterotrophic marine protists.

Thraustochytrids have recently emerged as a promising source for docosahexaenoic acid (DHA) production due to their high growth rate and oil content. In this study, two thraustochytrid isolates, Aurantiochytrium sp. PKU#SW7 and Thraustochytriidae sp. PKU#Mn16 were used for DHA production. Following growth parameters were optimized to maximize DHA production: temperature, pH, salinity, and glucose concentration. Both isolates achieved the highest DHA yield at the cultivation temperature of 28 °C, pH 6, 100 % seawater, and 2 % glucose. A DHA yield of 1.395 g/l and 1.426 g/l was achieved under the optimized culture conditions. Further investigation revealed that both isolates possess simple fatty acids profiles with palmitic acid and DHA as their dominant constituents, accounting for ∼79 % of total fatty acids. To date, very few studies have focused on the DHA distribution in various lipid fractions which is an important factor for identifying strains with a potential for industrial DHA production. In the present study, the lipids profiles of each strain both revealed that the majority of DHA was distributed in neutral lipids (NLs), and the DHA distribution in NLs of PKU#SW7 was exclusively in the form of triacylglycerols (TAGs) which suggest that PKU#SW7 could be utilized as an alternative source of DHA for dietary supplements. The fermentation process established for both strains also indicating that Aurantiochytrium sp. PKU#SW7 was more suitable for cultivation in fermenter. In addition, the high percentage of saturated fatty acids produced by the two thraustochytrids indicates their potential application in biodiesel production. Overall, our findings suggest that two thraustochytrid isolates are suitable candidates for biotechnological applications.

Growth and docosahexaenoic acid production performance of the heterotrophic marine microalgae Crypthecodinium cohnii in the wave‐mixed single‐use reactor CELL‐tainer

The process development of docosahexaenoic acid (DHA) with the heterotrophic marine microalgae Crypthecodinium cohnii is challenging, since the oxygen demand of the cells is high and the cell itself sensitive to high shear forces. Furthermore, the marine medium, which comprises a high salt concentration, accelerates corrosion of steel typically applied in bioreactors. Single‐use bioreactors (SUBs) provide an alternative to common stainless steel stirred tank reactors (STR) for the development of bioprocesses with marine microorganisms, when the development stage does not allow for the application of special steel or the replacement of chloride ions in the medium. Among the readily‐available SUBs, the CELL‐tainer exhibits outstanding high oxygen transfer rates. A direct comparison of this SUB with a standard stirred tank bioreactor (STR) was performed for the cultivation of C. cohnii. The biomass and DHA concentrations were similar in both devices, while the biomass yield coefficient, related to the glucose consumption, was reduced by 35% and the DHA yield coefficient by 21% in the STR. The impact of shear stress on cell physiology was elucidated by flow cytometry, revealing a five times higher amount of cell debris in the STR and 14 times higher amount of cells with a low cell membrane potential as evaluated by bis‐(1,3‐dibutylbarbituric acid) trimethine oxonol staining. Foam and biofilm formation, which is a problem in the STR, did not occur in the SUB. The study underlines that the CELL‐tainer can replace the STR, ensuring sufficient oxygen transfer rates and decreasing cell damage and foam formation.

Effects of temperature and carbon source on growth and docosahexaenoic acid accumulation of Crypthecodinium cohnii

Culture conditions were adapted to obtain larger biomass and higher docosahexaenoic acid(DHA)yield of Crypthecodinium cohnii under flask fermentation level.Results showed that the optimal culture should be at 22℃,in carbon-source culture with 12g/L glucose for 4d and then shifted to the same media but with 3g/L sodium acetate instead for 4d.Under this optimal condition,the C.cohnii biomass yield,DHA yield and DHA content reached 234.85mg/(L·d),17.45mg/(L·d),74.36mg/g dry weight respectively.

Culturable diversity and biochemical features of thraustochytrids from coastal waters of Southern China

Thraustochytrids are ubiquitous marine osmo-heterotrophic fungi-like microorganisms with only about 40 identified species till now. In this study, a total of 60 thraustochytrid strains were isolated from marine coastal habitats. Analysis of 18S rRNA gene sequences revealed that they belonged to three genera, i.e., Schizochytrium, Aurantiochytrium, and Thraustochytrium. All of the isolates were found to show considerable cellulolytic and lipolytic activities. Strains of Aurantiochytrium sp. and Thraustochytrium sp. were found to produce the highest levels of extracellular polysaccharides (EPS), which reached 345 μg ml(-1) in the growth media. Fourier transform infrared (FTIR) spectra of the EPS samples derived from two thraustochytrids (PKU#Sed1 and #SW1) displayed peaks for carbohydrates, proteins, lipids, uronic acids, and nucleic acids. Fatty acid profiles of four thraustochytrids comprised of palmitic acid (C16:0) and docosahexaenoic acid (DHA) as their major constituents. Schizochytrium sp. demonstrated the highest DHA production at 44 % of total fatty acids (TFA) with biomass and DHA yield of 7.1 and 1.6 g l(-1), respectively, on the fourth day of growth. All the four isolates exhibited considerable production of palmitic acid (16:0) in their fatty acid profiles ranging from 35 to 50 % TFA. This is the first report on extracellular enzymes, EPS, and DHA production from thraustochytrids isolated from the coastal habitats of China.

Regulation of docosahexaenoic acid production by Schizochytrium sp.: effect of nitrogen addition

Docosahexaenoic acid (DHA) percentage in total fatty acids (TFAs) is an important index in DHA microbial production. In this study, the change of DHA percentage in response to fermentation stages and the strategies to increase DHA percentage were investigated. Two kinds of conventional nitrogen sources, monosodium glutamate (MSG) and ammonium sulfate (AS), were tested to regulate DHA synthesis. Results showed that MSG addition could accelerate the substrate consumption rate but inhibit lipid accumulation, while AS addition could increase DHA percentage in TFAs effectively but extend fermentation period slightly. Finally, the AS addition strategy was successfully applied in 7,000-L fermentor and DHA percentage in TFAs and DHA yield reached 46.06% and 18.48g/L, which was 19.54 and 17.41% higher than that of no-addition strategy. This would provide guidance for the large-scale production of the other similar polyunsaturated fatty acid, and give insight into the nitrogen metabolism in oil-producing microorganisms.

Growth optimization of thraustochytrid strain 12B for the commercial production of docosahexaenoic acid

High yield of cell, lipid, and docosahexaenoic acid (DHA) from thraustochytrid strain 12B were achieved without the use of a complex medium and at low NaCl concentration which is detrimental to avoid unnecessary corrosion of steel tank equipment during cultivation. Culture medium that contained only 0.1% NaCl and 1% MgSO4 in an organic base solution containing 8% glucose, 1% yeast extract, and 1% peptone, referred here as NM medium, was found to be as good as or superior to the culture medium prepared from 50%(v/v) seawater with percentage lipid/dry cell weight (DCW) of 66.4%(w/w) and DHA yield up to 43.95 mg/g DCW for the thraustochytrid strain 12B. The NM medium was also applicable to the prominently high DHA-accumulating Schizochytrium limacinum SR21, and therefore this medium could probably be used for other thraustochytrid and other types of microbial strains as well.

Isolation and characterization of Aurantiochytrium species: high docosahexaenoic acid (DHA) production by the newly isolated microalga, Aurantiochytrium sp. SD116

A heterotrophic microalga, strain SD116, with the ability to produce high concentrations of docosahexaenoic acid (DHA, C22:6n-3) was isolated from Shuidong Bay, Guangdong Province, China. Nucleotide sequence analysis of the 18S rDNA of SD116 showed that the strain has a close phylogenetic relationship to Aurantiochytrium species. The highest rates for growth and DHA accumulation for SD116 were obtained in 6.0% glucose, 2.0% yeast extract, and 50% artificial seawater (ASW) at a pH of 7 at 28°C. The maximum total lipid content reached 56.3% of the dry cell weight (DCW), and the maximum DHA content accounted for 50.9% of the total fatty acid (TFA) content. It was further found that urea may be a potential nitrogen source for industrial fermentation because of its cheap price and ability to induce a relatively high biomass and lipid production capacity. Using 5 L fermenters, the DCW, total lipid content, and DHA yield were found to be 70.43 g L(-1), 71.09% of the DCW, and 17.42 g L(-1) (34.79% of the TFA), respectively. The results show that Aurantiochytrium sp. SD116 is a promising candidate for commercial DHA production and could be useful for the synthesis of biomass-related products.

Utilization of Waste Syrup for Production of Polyunsaturated Fatty Acids and Xanthophylls by Aurantiochytrium

In the food industry, syrups containing a high concentration of sugar used for fruit preservation is abundantly discharged as a food processing waste and disposed by incineration, resulting in the rise of the manufacturing cost and environmental pollution. This study demonstrates how waste syrup can be utilized as carbon source for production of docosahexaenoic acid (DHA) and astaxanthin by the thraustochytrid strain, Aurantiochytrium sp. KH105. The strain could grow in culture medium containing 3-50% waste syrup, and the maximum yields of DHA and astaxanthin were 207.6 mg/L (at 50%) and 1.1 mg/L (at 25%), respectively. After the optimization of culture medium composition by response surface method, DHA and astaxanthin yields increased by 2.1 and 1.5 fold, respectively. When the waste syrup was treated with activated charcoal, citrate concentration in the syrup was reduced and the astaxanthin yield increased by 2.3 fold. This study shows that the waste syrup can be effectively used for the functional lipid production by the thraustochytrid.

A novel fed-batch process based on the biology of Aurantiochytrium sp. KRS101 for the production of biodiesel and docosahexaenoic acid

The biology of Aurantiochytrium sp. KRS101 was thoroughly investigated to enhance its production of biodiesel and docosahexaenoic acid (DHA). Nutrients and salinity were optimized to prevent biomass loss due to cell rupture. Calculation of yield coefficients showed that nitrogen was mostly responsible for the early stage of cell growth or division, whereas carbon was necessary for the entire process of cell development, particularly cell enlargement during late stages. Using these distinctive yield coefficients, a modified fed-batch cultivation method was designed, resulting in increases in palmitic acid (PA) and DHA production of up to 137% and 29%, respectively. This modified fed-batch cultivation method, using appropriate supplies of nitrogen and carbon, may improve the yields of PA and DHA, thus expanding the biotechnological applications of Aurantiochytrium sp. KRS101.

Impact of phosphate concentration on docosahexaenoic acid production and related enzyme activities in fermentation of Schizochytrium sp.

Docosahexaenoic acid (DHA) is an important and widely used infant food additive. In this study, the effects of phosphate concentration on lipid and especially DHA synthesis in the oleaginous fungi Schizochytrium sp. HX-308 have been investigated in batch cultures. The maximum DHA yield (8.9g/L) and DHA productivity (148.3mg/L h) in 0.1g/L KH2PO4 concentration were higher than the DHA yield (6.2g/L) and DHA productivity (86.1mg/L h) in 4g/L KH2PO4 concentration. Furthermore, differences in related enzyme activities (malic enzyme, glucose-6-phosphate dehydrogenase and NAD(+)-isocitrate dehydrogenase) between phosphate-sufficient and phosphate-limitation conditions were assayed. The results showed that the phosphate-limitation condition could maintain higher activities of malic enzyme and glucose-6-phosphate dehydrogenase in addition to lower activity of NAD(+)-isocitrate dehydrogenase. In addition, glucose-6-phosphate dehydrogenase might be the main supplier of NADPH at the early stage of fermentation while malic enzyme might be the provider at the late stage. This information might explain the regulation mechanism of phosphate limitation for lipid production and be useful for further DHA production enhancement.

Efficient Display of Active Geotrichum sp. Lipase on Pichia pastoris Cell Wall and Its Application as a Whole-Cell Biocatalyst To Enrich EPA and DHA in Fish Oil

Geotrichum sp. lipase (GSL) was first displayed on the cell wall of Pichia pastoris on the basis of the a-agglutinin anchor system developed in Saccharomyces cerevisiae . Surface display levels were monitored using Western blotting, immunofluorescence miscroscopy, and fluorescence-activated cell sorting analysis. Lipase activity of the yeast whole cells reached a maximum at 273 ± 2.4 U/g of dry cells toward olive oil after 96 h of culture at 30 °C, with optimal pH and temperature at 7.5 and 45 °C, respectively. Displayed GSL exhibited relatively high stability between pH 6.0 and 8.0 and retained >70% of the maximum activity. The surface-displayed lipase retained 80% of its original activity after incubation at 45 °C for 4 h. Moreover, the GSL-displaying yeast whole cells were then used as a biocatalyst to enrich eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) from fish oil on the basis of selective hydrolysis. As a result, EPA and DHA increased from 1.53 and 24.1% in the original fish oil to 1.85 and 30.86%, which were increases of 1.21- and 1.29-fold, respectively. The total yield of EPA and DHA reached 46.62%.

Use of algae or algal oil rich in n-3 fatty acids as a feed supplement for dairy cattle

Fish oil is used as a ration additive to provide n-3 fatty acids to dairy cows. Fish do not synthesize n-3 fatty acids; they must consume microscopic algae or other algae-consuming fish. New technology allows for the production of algal biomass for use as a ration supplement for dairy cattle. Lipid encapsulation of the algal biomass protects n-3 fatty acids from biohydrogenation in the rumen and allows them to be available for absorption and utilization in the small intestine. Our objective was to examine the use of algal products as a source for n-3 fatty acids in milk. Four mid-lactation Holsteins were assigned to a 4×4 Latin square design. Their rations were supplemented with 1× or 0.5× rumen-protected (RP) algal biomass supplement, 1× RP algal oil supplement, or no supplement for 7 d. Supplements were lipid encapsulated (Balchem Corp., New Hampton, NY). The 1× supplements provided 29g/d of docosahexaenoic acid (DHA), and 0.5× provided half of this amount. Treatments were analyzed by orthogonal contrasts. Supplementing dairy rations with rumen-protected algal products did not affect feed intake, milk yield, or milk component yield. Short- and medium-chain fatty acid yields in milk were not influenced by supplements. Both 0.5× and 1× RP algae supplements increased daily milk fat yield of DHA (0.5 and 0.6±0.10g/d, respectively) compared with 1× RP oil (0.3±0.10g/d), but all supplements resulted in milk fat yields greater than that of the control (0.1±0.10g/d). Yield of trans-18:1 fatty acids in milk fat was also increased by supplementation. Trans-11 18:1 yield (13, 20, 27, and 15±3.0g/d for control, 0.5× RP algae, 1× RP algae, and 1× RP oil, respectively) was greater for supplements than for control. Concentration of DHA in the plasma lipid fraction on d 7 showed that the DHA concentration was greatest in plasma phospholipid. Rumen-protected algal biomass provided better DHA yield than algal oil. Feeding lipid-encapsulated algae supplements may increase n-3 content in milk fat without adversely affecting milk fat yield; however, preferential esterification of DHA into plasma phospholipid may limit its incorporation into milk fat.

The effects of culture condition on the growth property and docosahexaenoic acid production from Thraustochytrium aureum ATCC 34304

The effect of the composition of artificial sea water (ASW) medium on the growth properties and docosahexaenoic acid (DHA) production from Thraustochytrium aureum ATCC 34304 was investigated. A maximum dry cell weight (DCW) of 3.71 g/l was obtained when the NaCl and MgSO4 concentration in the ASW media were 15 and 0 g/l, respectively. The maximum initial specific growth (ISG) rate of 0.16 was attained at 7.5 g/l NaCl and 18 g/l MgSO4, while the minimum ISG rate (0.02) was obtained at 5.0 g/l NaCl and 4.5 g/l MgSO4. The least doubling time required for biomass production was 4.3 h at 7.5 g/l NaCl and different MgSO4 concentrations. A maximum of 7.9 g/l DCW was obtained on the fourth day of cultivation at 30 g/l glucose and 2.5 g/l (each) yeast extract (YE) and peptone. The DHA content in the lipids was significantly affected by the concentration of glucose and nitrogen sources (YE and peptone) in the ASW medium. At the lowest glucose (10 g/l) and YE/peptone (0.5 g/l) concentration and highest glucose (30 g/l) and YE/peptone (2.5 g/l) concentration, the DHA content was 34.725 and 40.33%, respectively, relative to total lipid content. However, the DHA content in the lipid was not affected by the NaCl and MgSO4 concentration. At the lowest NaCl (2.5 g/l) and MgSO4 (4.5 g/l) concentration and highest NaCl (60 g/l) and MgSO4 (18 g/l) concentration the DHA content was 39.62 and 38.48%, respectively. The maximum DHA content in the lipid was 49.01% after four days of cultivation when 7.5 g/l NaCl and 4.5 g/l MgSO4 were in the ASW medium. The growth properties of T. aureum ATCC 34304 for biomass production and DHA yield in the lipid content were found to be affected by NaCl and glucose concentration.