Lipid production by microalgae <Emphasis Type="Italic">Chlorella protothecoides</Emphasis> with volatile fatty acids (VFAs) as carbon sources in heterotrophic cultivation and its economic assessment

Volatile fatty acids (VFAs) that can be derived from food wastes were used for microbial lipid production by Chlorella protothecoides in heterotrophic cultures. The usage of VFAs as carbon sources for lipid accumulation was investigated in batch cultures. Culture medium, culture temperature, and nitrogen sources were explored for lipid production in the heterotrophic cultivation. The concentration and the ratio of VFAs exhibited significant influence on cell growth and lipid accumulation. The highest lipid yield coefficient and lipid content of C. protothecoides grown on VFAs were 0.187 g/g and 48.7 %, respectively. The lipid content and fatty acids produced using VFAs as carbon sources were similar to those seen on growth and production using glucose. The techno-economic analysis indicates that the biodiesel derived from the lipids produced by heterotrophic C. protothecoides with VFAs as carbon sources is very promising and competitive with other biofuels and fossil fuels.     

Effects of molten-salt/ionic-liquid mixture on extraction of docosahexaenoic acid (DHA)-rich lipids from Aurantiochytrium sp. KRS101

In this study, lipid extraction from Aurantiochytrium sp. was performed using a molten-salt/ionic-liquid mixture. The total fatty acid content of Aurantiochytrium sp. was 478.8 mg/g cell, from which 145 mg/g cell (30.3 % of total fatty acids) of docosahexaenoic acid (DHA) was obtained. FeCl₃·6H₂O showed a high lipid extraction yield (207.9 mg/g cell), when compared with that of [Emim]OAc, which was only 118.1 mg/g cell; notably however, when FeCl₃·6H₂O was mixed with [Emim]OAc (5:1, w/w), the yield was increased to 478.6 mg/g cell. When lipid was extracted by the FeCl₃·6H₂O/[Emim]OAc mixture at a 5:1 (w/w) blending ratio under 90 °C, 30 min reaction conditions, the fatty acid content of the extracted lipid was a high purity 997.7 mg/g lipid, with most of the DHA having been extracted (30.2 % of total fatty acids). Overall, lipid extraction from Aurantiochytrium sp. was enhanced by the synergistic effects of the molten-salt/ionic-liquid mixture with different ions.     

Cultivation of <Emphasis Type="Italic">Chlorella protothecoides</Emphasis> in anaerobically treated brewery wastewater for cost-effective biodiesel production

The use of wastewater has been investigated to overcome the economic challenge involved with a production of microalgae-based biodiesel. In this study, to achieve economical biodiesel production along with effective wastewater treatment at the same time, anaerobically treated brewery wastewater (ABWW) was utilized as a low-cost nutrient source, in the cultivation of Chlorella protothecoides. About 96 and 90 % of total nitrogen and phosphorus in ABWW were removed, respectively, while C. protothecoides was accumulating 1.88 g L^?1 of biomass. The C. protothecoides grown in ABWW showed increases in cell size and cell aggregation, resulting in a near 80 % enhanced harvesting efficiency within 20 min, as compared with only 4 % in BG-11. In addition, the total fatty acid content of the C. protothecoides grown in ABWW increased by 1.84-fold (35.94 ± 1.54 % of its dry cell weight), relative to that of BG-11.     

Carotenoid and lipid production by the autotrophic microalga Chlorella protothecoides under nutritional, salinity, and luminosity stress conditions

Today microalgae represent a viable alternative source for high-value products. The specie Chlorella protothecoides (Cp), heterotrophically grown, has been widely studied and provides a high amount of lutein and fatty acids (FA) and has a good profile for biodiesel production. This work studies carotenoid and FA production by autotrophic grown Cp. Cp was grown until the medium’s nitrogen was depleted, then diluted in NaCl solution, resulting in nutritional, luminosity, and salinity stresses. Different NaCl concentrations were tested (10, 20, 30 g/L) at two different dilutions. After dilution, a color shifting from green to orange-red was noticed, showing carotenoid production. The best production of both carotenoids and FA was attained with a 20 g/L NaCl solution. The total carotenoid content was 0.8 % w/w (canthaxanthin (23.3 %), echinenone (14.7 %), free astaxanthin (7.1 %), and lutein/zeaxanthin (4.1 %)). Furthermore, the total lipid content reached 43.4 % w/w, with a FA composition of C18:1 (33.64 %), C16:0 (23.30 %), C18:2 (11.53 %), and less than 12 % of C18:3, which is needed to fulfill the biodiesel quality specifications (EN 14214).     

Boosting fatty acid synthesis in <Emphasis Type="Italic">Rhodococcus opacus</Emphasis> PD630 by overexpression of autologous thioesterases

Objectives

To explore the role of thioesterases in Rhodococcus opacus PD630 by endogenously overexpression in this bacteria for increased lipid production.

Results

Overexpression of four thioesterases from R. opacus PD630 in E. coli led to a 2- to 8-fold increase in C16:1 and C18:1 fatty acids while, when overexpressed in R. opacus PD630, only two recombinants had significant effect on the quantities and compositions of total fatty acid. The contents of total fatty acids (FAs) in two recombinants, pJTE2 (OPAG_00508 thioesterase) and pJTE4 (WP_012687673.1 thioesterase), were 400–460 mg/g (CDW) which is 1.5 times of wild-type strain PD630 (300-350 mg/g CDW), and 20–30 % (w/w) more than that of the control strain PDpJAM2 (330-370 mg/g CDW). The contents of 17:1 and 18:1 fatty acids increased by about 27 and 35 %, respectively, in pJTE2 and by 35 and 20 %, respectively, in pJTE4 compared with the control strain.

Conclusions

The engineered strains showed improved production of lipid (as total fatty acids), and could also tailor the composition of the fatty acid profile when cultured in mineral salts medium using glucose as sole carbon source.

     

High cell density cultivation of a novel Aurantiochytrium sp. strain TC 20 in a fed-batch system using glycerol to produce feedstock for biodiesel and omega-3 oils

A recently isolated Australian Aurantiochytrium sp. strain TC 20 was investigated using small-scale (2 L) bioreactors for the potential of co-producing biodiesel and high-value omega-3 long-chain polyunsaturated fatty acids. Higher initial glucose concentration (100 g/L compared to 40 g/L) did not result in markedly different biomass (48 g/L) or fatty acid (12–14 g/L) yields by 69 h. This comparison suggests factors other than carbon source were limiting biomass production. The effect of both glucose and glycerol as carbon sources for Aurantiochytrium sp. strain TC 20 was evaluated in a fed-batch process. Both glucose and glycerol resulted in similar biomass yields (57 and 56 g/L, respectively) by 69 h. The agro-industrial waste from biodiesel production—glycerol—is a suitable carbon source for Aurantiochytrium sp. strain TC 20. Approximately half the fatty acids from Aurantiochytrium sp. strain TC 20 are suitable for development of sustainable, low emission sources of transportation fuels and bioproducts. To further improve biomass and oil production, fortification of the feed with additional nutrients (nitrogen sources, trace metals and vitamins) improved the biomass yield from 56 g/L (34 % total fatty acids) to 71 g/L (52 % total fatty acids, cell dry weight) at 69 h; these yields are to our knowledge around 70 % of the biomass yields achieved, however, in less than half of the time by other researchers using glycerol and markedly greater than achieved using other industrial wastes. The fast growth and suitable fatty acid profile of this newly isolated Aurantiochytrium sp. strain TC 20 highlights the potential of co-producing the drop-in biodiesel and high value omega-3 oils.     

Phosphate limitation promotes unsaturated fatty acids and arachidonic acid biosynthesis by microalgae <Emphasis Type="Italic">Porphyridium purpureum</Emphasis>

Polyunsaturated fatty acids (PUFAs) are highly appreciated on their nutritive value for human health and aquaculture. P. purpureum, one of the red microalgae acknowledged as a promising accumulator of ARA, was chosen as the target algae in the present research. Effects of sodium bicarbonate (0.04–1.2 g/L), temperature (25, 30 and 33 °C) and phosphate (0.00–0.14 g/L) on biomass yield, total fatty acids (TFA) and arachidonic acid (ARA) accumulation were investigated systemically. NaHCO₃ dose of 0.8 g/L and moderate temperature of 30 °C were preferred. In addition, TFA and ARA production were significantly enhanced by an appropriate concentration of phosphate, and the highest TFA yield of 666.38 mg/L and ARA yield of 159.74 mg/L were obtained at a phosphate concentration of 0.035 g/L. Interestingly, with phosphate concentration continuing to fall, UFA/TFA and ARA/EPA ratios were increased accordingly, suggesting that phosphate limitation promoted unsaturated fatty acids and arachidonic acid biosynthesis. Low concentration of phosphate may be favored to increase the enzymatic activities of ?6-desaturase, which played a key role in catalyzing the conversion of C16:0 to C18:2, and thus the selectivity of UFA increased. Meanwhile, the increase of ARA selectivity could be attributed to ω6 pathway promotion and ?17-desaturase activity inhibition with phosphate limitation. Phosphate limitation strategy enhanced unsaturated fatty acids and ARA biosynthesis in P. purpureum, and can be applied in commercial scale manufacturing and commercialization of ARA.     

High-productivity lipid production using mixed trophic state cultivation of <Emphasis Type="Italic">Auxenochlorella</Emphasis> (<Emphasis Type="Italic">Chlorella</Emphasis>) <Emphasis Type="Italic">protothecoides</Emphasis>

A mixed trophic state production process for algal lipids for use as feedstock for renewable biofuel production was developed and deployed at subpilot scale using a green microalga, Auxenochlorella (Chlorella) protothecoides. The process is composed of two separate stages: (1) the photoautotrophic stage, focused on biomass production in open ponds, and (2) the heterotrophic stage focused on lipid production and accumulation in aerobic bioreactors using fixed carbon substrates (e.g., sugar). The process achieved biomass and lipid productivities of 0.5 and 0.27 g/L/h that were, respectively, over 250 and 670 times higher than those obtained from the photoautotrophic cultivation stage. The biomass oil content (over 60 % w/DCW) following the two-stage process was predominantly monounsaturated fatty acids (~82 %) and largely free of contaminating pigments that is more suitable for biodiesel production than photosynthetically generated lipid. Similar process performances were obtained using cassava hydrolysate as an alternative feedstock to glucose.     

Euglena sp. as a suitable source of lipids for potential use as biofuel and sustainable wastewater treatment

Prolific algal growth in sewage ponds with high organic loads in the tropical regions can provide cost-effective and efficient wastewater treatment and biofuel production. This work examines the ability of Euglena sp. growing in wastewater ponds for biofuel production and treatment of wastewater. The algae were isolated from the sewage treatment plants and were tested for their nutrient removal capability. Compared to other algae, Euglena sp. showed faster growth rates with high biomass density at elevated concentrations of ammonium nitrogen (NH₄-N) and organic carbon (C). Profuse growth of these species was observed in untreated wastewaters with a mean specific growth rate (μ) of 0.28 day^?1 and biomass productivities of 132 mg ?L^?1? day^?1. The algae cultured within a short period of 8 days resulted in the 98 % removal of NH₄-N, 93 % of total nitrogen 85 % of ortho-phosphate, 66 % of total phosphate and 92 % total organic carbon. Euglenoids achieved a maximum lipid content of 24.6 % (w/w) with a biomass density of 1.24 g ?L^?1 (dry wt.). Fourier transform infrared spectra showed clear transitions in biochemical compositions with increased lipid/protein ratio at the end of the culture. Gas chromatography and mass spectrometry indicated the presence of high contents of palmitic, linolenic and linoleic acids (46, 23 and 22 %, respectively), adding to the biodiesel quality. Good lipid content (comprised quality fatty acids), efficient nutrient uptake and profuse biomass productivity make the Euglena sp. as a viable source for biofuel production in wastewaters.     

Olfactory responses of Epilachna dodecastigma (Coleoptera: Coccinellidae) to long-chain fatty acids from Momordica charantia leaves

Extraction, thin-layer chromatography and gas chromatography–mass spectrophotometry analyses revealed the presence of 12, 13, and 12 fatty acids in young, mature, and senescent leaves of Momordica charantia L., representing 87.30, 95.25, and 83.11 % of the total fatty acids, respectively. The proportion of saturated fatty acids was highest in senescent leaves (78.60 %) followed by young leaves (69.42 %) and mature leaves (48.92 %), with the balance accounted for by unsaturated fatty acids. Palmitic acid was the predominant saturated fatty acid in the three types of leaves, whereas alpha-linolenic acid was the predominant unsaturated fatty acid. The fatty acids from young, mature, and senescent leaves followed by the application of a synthetic mixture of fatty acids that was comparable to the natural fatty acids found in the three types of leaves, elicited the attraction of the female insect Epilachna dodecastigma (Coleoptera: Coccinellidae) at 50–200, 50–200, and 100–200 μg/ml concentrations, respectively, in a Y-shaped glass tube olfactometer bioassay. Individual synthetic fatty acids were also evaluated by the olfactometer bioassay at concentrations comparable to the proportions detected in the three types of leaves. Individual synthetic palmitic acid, stearic acid, oleic acid, linoleic acid, and alpha-linolenic acid at 58.24, 13.96, 29.40, 30.31, and 29.76 μg, respectively, attracted the insect. A synthetic blend of 79.13, 10.57, 29.40, 30.31, and 36.33 μg of palmitic, stearic, oleic, linoleic, and alpha-linolenic acids, respectively, which is the proportion present in a 200 μg/ml concentration of fatty acids of mature leaves, or of 116.49, 13.96, and 29.76 μg of palmitic, stearic and alpha-linolenic acids, respectively, which is the proportion present in a 200 μg/ml concentration of natural fatty acids of young leaves, served as attractants for E. dodecastigma.     

Effects of two sodium salts on fatty acid and essential oil composition of basil (Ocimum basilicum L.) leaves

The effects of two sodium salts on growth, fatty acids, and essential oil compositions were investigated in a medicinal and aromatic plant, Ocimum basilicum cultivated in hydroponic medium. Plants were subjected to an equimolar concentration of Na₂SO₄ (25 mM) and NaCl (50 mM) for 15 days. Our results showed that leaf growth rate was more depressed by 25 mM Na₂SO₄ than by 50 mM NaCl. The total fatty acid contents did not show any change in plants. α-Linolenic, palmitic, and linoleic acids were the major fatty acids. The identification of basil leaf fatty acids has not been previously studied and this work revealed the predominance of polyunsaturated fatty acids. Under both salts, leaf fatty acid composition remained unchanged. Regarding the essential oil yield, it decreased significantly by 28 % under 25 mM Na₂SO₄ and showed an increase by 27 % under 50 mM NaCl. The major volatile compound in leaves was linalool with 34.3 % of total essential oil constituents, followed by eugenol (19.8 %), 1.8-cineole (14.4 %) and methyl eugenol (5.2 %). Further, levels of eugenol and methyl eugenol were most modulated by salt, and the negative correlation between these two compounds reflects the stimulation of O-methyltransferase activity under both salts.     

Agitated shoot cultures of <Emphasis Type="Italic">Aronia arbutifolia</Emphasis> and <Emphasis Type="Italic">Aronia</Emphasis> × <Emphasis Type="Italic">prunifolia</Emphasis>: biotechnological studies on the accumulation of phenolic compounds and biotransformation capability

In methanolic extracts of the biomass from agitated cultures of Aronia arbutifolia and Aronia?×?prunifolia grown on four variants of the Murashige and Skoog (MS) medium, with different concentrations of plant growth regulators (PGRs): BA and NAA (0.5–3.0 mg/l), the quantities of phenolic acids (19 compounds) and flavonoids (11 compounds) were estimated using the LC-DAD method. The amounts of individual metabolites and total contents were dependent on the concentration of PGRs in MS medium variant. The maximum total amounts of phenolic acids and flavonoids reached 360.80 and 65.26 mg/100 g DW, and 659.51 and 78.34 mg/100 g DW for A. arbutifolia and A. × prunifolia, respectively. The main metabolites in the biomass of both plants were chlorogenic acid, rosmarinic acid and quercitrin (max. 175.94, 147.98 and 41.14 mg/100 g DW, and 260.34, 225.26 and 78.34 mg/100 g DW, respectively). The cells of both plants convert the exogenously supplied hydroquinone into its β-D-glucoside—arbutin. The maximal total content of the product accumulated in the biomass and media reached 83.55 and 73.62 mg/g DW. The obtained results demonstrated for the first time a high biosynthetic potential of agitated cultures of both plants.     

Sophorolipid production from delignined corncob residue by Wickerhamiella domercqiae var. sophorolipid CGMCC 1576 and Cryptococcus curvatus ATCC 96219

Delignined corncob residue hydrolysate (DCCRH) and detoxified DCCRH were used for single cell oil (SCO) and single cell protein (SCP) production of Cryptococcus curvatus ATCC 96219 and for sophorolipid (SL) production of Wickerhamiella domercqiae var. sophorolipid CGMCC 1576. Both C. curvatus and W. domercqiae could utilize glucose in DCCRH to grow and accumulate lipids or particle-shaped SLs. DCCRH detoxification by activated carbon adsorption not only improved cell growth and lipid accumulation of C. curvatus but also increased SL production and proportion of lactonic SL in total SL. A total biomass of 17.36 g/l with a lipid content of 44.36 % could be achieved after cultivation of C. curvatus on the detoxified DCCRH. The predominant fatty acids of the produced SCO were oleic, stearic, and palmitic acids (27.2, 20.5, and 15.7 %, respectively). When W. domercqiae cells were cultivated on DCCRH and SCO, total SL production of 39.08 g/l (DCCRH?+?SCO) and 42.06 g/l (detoxified DCCRH?+?SCO) were obtained. Furthermore, when cell lysate of C. curvatus, oleic acid, and DCCRH/detoxified DCCRH was used as nitrogen and carbon sources, total SL production reached 37.19 g/l and 48.97 g/l, respectively. These results demonstrated that renewable DCCRH can be utilized for the production of high-value SCO and SLs.     

Lipid and total fatty acid productivity in photoautotrophic fresh water microalgae: screening studies towards biodiesel production

Microalgae are considered as a promising feedstock for biomass production. The selection of the most suitable species is based on several key parameters such as lipid and fatty acid productivity. In the present study, the growth of different microalgae strains was examined in freshwater media for photoautotrophs suited for large-scale applications to identify the most suitable medium for each species. In the optimal medium, Scenedesmus obliquus showed the highest biomass productivity measured as increase of cell dry weight (0.25 g cellu dry weight (CDW) L^?1 day^?1), while Botryococcus braunii showed the highest lipid and total fatty acid content (430 and 270 mg g^?1 CDW, respectively) among the tested species. Regarding lipid and total fatty acid productivity, S. obliquus was the most lipid and total fatty acid productive strain with 41 and 18 mg L^?1 day^?1 during the exponential phase, respectively. Additionally, the proportion of saturated and monounsaturated fatty acids increased with duration of the incubation in S. obliquus, while polyunsaturated fatty acids decreased. These results nominate S. obliquus as a promising microalga in order to serve as a feedstock for renewable energy production.     

Screening and characterization of Isochrysis strains and optimization of culture conditions for docosahexaenoic acid production

Isochrysis is a genus of marine unicellular microalgae that produces docosahexaenoic acid (DHA, C22:6), a very long chain polyunsaturated fatty acid (PUFA) of significant health and nutritional value. Mass cultivation of Isochrysis for DHA production for human consumption has not been established due to disappointing low DHA productivity obtained from commonly used Isochrysis strains. In this study, 19 natural Isochrysis strains were screened for DHA yields and the results showed that the cellular DHA content ranged from 6.8 to 17.0 % of total fatty acids with the highest DHA content occurring in the exponential growth phase. Isochrysis galbana #153180 exhibited the greatest DHA production potential and was selected for further investigation. The effects of different light intensities, forms, and concentrations of nitrogen, phosphorus, and salinity on growth and DHA production of I. galbana #153180 were studied in a bubble column photobioreactor (PBR). Under favorable culture conditions, I. galbana #153180 contained DHA up to 17.5 % of total fatty acids or 1.7 % of cell dry weight. I. galbana #153180 was further tested in outdoor flat-plate PBRs varying in light path length, starting cell density (SCD), and culture mode (batch versus semicontinuous). When optimized, record high biomass and DHA productivity of I. galbana #153180 of 0.72 g L^?1?day^?1 and 13.6 mg?L^?1?day^?1, or 26.4 g?m^?2?day^?1 and 547.7 mg?m^?2?day^?1, respectively, were obtained, suggesting that I. galbana #153180 may be a desirable strain for commercial production of DHA.     

Polyunsaturated fatty acid production by marine bacteria

Polyunsaturated fatty acids are important in maintaining human health. Limitations associated with current sources of ω-3 fatty acids and ω-6 fatty acids, from animal and plant sources, have led to increased interest in microbial production. Marine bacteria may provide a suitable alternative, although the isolation of production strains and the identification of operating conditions must be addressed before manufacturing processes become economically viable. Marine isolate 560 was identified as an eicosapentaenoic acid (EPA) producer via GC/MS. The isolate was initially identified as Vibrio cyclitrophicus by 16S rRNA sequencing. Statistically based experimental designs were applied to the optimisation of medium components and environmental factors for the production of EPA. A Plackett–Burman design was used to screen for the effect of temperature, pH, and media components. Subsequently, the concentrations of NaCl, yeast extract, and peptone, identified as significant factors, were optimised using a central composite design. The predicted optimal combination of media components for maximum EPA production (4.8 mg/g dry weight) was determined as 7.9 g/l peptone, 16.2 g/l NaCl, and 6.2 g/l yeast extract. On transfer of this process to bioreactor cultivation, where a range of pH and DO values were tested, the maximum amount of EPA produced increased to 7.5 mg/g dry weight and 10 % of the total fatty acid.     

Bacterial Diversity and Bioprospecting for Cold-Active Lipases,Amylases and Proteases,from Culturable Bacteria of Kongsfjorden and Ny-Ålesund,Svalbard, Arctic

Culturable bacterial diversity of seven marine sediment samples of Kongsfjorden and a sediment and a soil sample from Ny-Ålesund, Svalbard, Arctic was studied. The bacterial abundance in the marine sediments of Kongsfjorden varied marginally (0.5 × 10³–1.3 × 10⁴ cfu/g sediment) and the bacterial number in the two samples collected from the shore of Ny-Ålesund also was very similar (0.6 × 10⁴ and 3.4 × 10⁴, respectively). From the nine samples a total of 103 bacterial isolates were obtained and these isolates could be grouped in to 47 phylotypes based on the 16S rRNA gene sequence belonging to 4 phyla namely Actinobacteria, Bacilli, Bacteroidetes and Proteobacteria. Representatives of the 47 phylotypes varied in their growth temperature range (4–37°C), in their tolerance to NaCl (0.3–2 M NaCl) and growth pH range (2–11). Representatives of 26 phylotypes exhibited amylase and lipase activity either at 5 or 20°C or at both the temperatures. A few of the representatives exhibited amylase and/or lipase activity only at 5°C. None of the phylotypes exhibited protease activity. Most of the phylotypes (38) were pigmented. Fatty acid profile studies indicated that short chain fatty acids, unsaturated fatty acids, branched fatty acids, the cyclic and the cis fatty acids are predominant in the psychrophilic bacteria.     

Magnetophoretic harvesting of freshwater microalgae using polypyrrole/Fe<Subscript>3</Subscript>O<Subscript>4</Subscript> nanocomposite and its reusability

A magnetophoretic harvesting agent, a polypyrrole/Fe₃O₄ magnetic nanocomposite, is proposed as a cost and energy efficient alternative to recover biomass of the microalgae Botryococcus braunii, Chlorella protothecoides, and Chlorella vulgaris from their culture media. The maximal recovery efficiency reached almost 99 % for B. braunii, 92.4 % for C. protothecoides, and 90.8 % for C. vulgaris. The maximum adsorption capacity (Q ₀) of the magnetic nanocomposite for B. braunii (63.49 mg dry biomass mg^?1 PPy/Fe₃O₄) was higher than that for C. protothecoides (43.91 mg dry biomass mg^?1 PPy/Fe₃O₄) and C. vulgaris (39.98 mg dry biomass mg^?1 PPy/Fe₃O₄). The highest harvesting efficiency for all the studied microalgae were at pH 10.0, and measurement of zeta-potential confirmed that the flocculation was induced by charge neutralization. This study showed that polypyrrole/Fe₃O₄ can be a promising flocculant due to its high efficacy, low dose requirements, short settling time, its integrity with cells, and with great potential for saving energy because of its recyclability.     

Assessment of the antimicrobial,antioxidant and cytotoxic activities of the wild edible mushroom <Emphasis Type="Italic">Agaricus lanipes</Emphasis> (F.H. Møller &amp; Jul. Schäff.) Hlaváček

The present investigation was undertaken to evaluate the antimicrobial and antioxidant activities of the wild edible mushroom Agaricus lanipes, and also to investigate its cytotoxicity and potential and possible apoptotic effect against the A549 lung cancer cell line in in vitro conditions. Total antioxidant capacity, total phenolic content, total oxidant status, total antioxidant status, lipid hydroperoxides, and total free –SH levels of A. lanipes were found to be 4.55 mg T/g, 14.6 mg GA equivalent/g, 3.10 mg H₂O₂ equivalent/g, 2.25 mg H₂O₂ equivalent/g, and 1.90 µmol/g, respectively. The methanolic extract of A. lanipes had relatively strong antimicrobial activity against seven tested microorganism strains. It also had high anti-proliferative potency and strong pro-apoptotic effects, and this mushroom used as a daily nutrient could be a source for new drug developments and treatment in cancer therapies, and could be a guide for studies in this area.     

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.