Search Results (53)

Aurantiochytrium sp., a heterotrophic microorganism, has received increasing attention for its high production of polyunsaturated fatty acids and has been widely applied in various industries. This study intended to optimize the carotenoid synthesis pathway in Aurantiochytrium sp. by metabolic engineering to increase the carotenoid content. Multi-sourced key enzyme genes involved in lipid synthesis (LPAAT and DGAT) and astaxanthin synthesis (crtZ and crtW) were selected to construct single-gene expression vectors and transformed into Aurantiochytrium sp. The results showed that the overexpression of LPAAT of Phaeodactylum tricornutum in Aurantiochytrium sp. caused an increase of 39.3% in astaxanthin, 424.7% in β-carotene, 901.8% in canthaxanthin, and 575.9% in lutein, as well as a down-regulation of 15.3% in the fatty acid content. Transcriptomics analysis revealed enhanced expression of genes involved in purine and amino acid metabolism in the transformed strains, and the down-regulation of the citric acid cycle led to an increase in the source of acetyl coenzyme A for the production of fatty acids. This study provides strong experimental evidence to support the application of increasing carotenoid levels in Aurantiochytrium sp. Full article

This study investigated a sustainable alternative to squalene production utilizing Thraustochytrium sp. and Aurantiochytrium sp., thereby reducing dependence on critically endangered sharks exploited for this compound. By optimizing fed-batch cultivation, a technique prevalent in industrial biotechnology, we have enhanced squalene yields and have demonstrated, through sensitivity analysis, the significance of this shift in preserving species at risk of extinction. Optimization of culture conditions led to the highest biomass concentrations for Thraustochytrium sp. being achieved at lower C–N ratios (<5.0), while the optimal biomass production for Aurantiochytrium sp. occurred in culture media with a high C–N ratio of 54:50. Regarding squalene production, Thraustochytrium sp. produced 26.13 mg/L in the fed-batch system after 72 h, and Aurantiochytrium sp. produced 54.97 mg/L in a batch system with 30 g/L glucose and 0.22 g/L nitrogen after 96 h, showcasing their potential for industrial applications. Moreover, the sensitivity analysis revealed that, on an industrial scale, both strains could produce up to 59.50 t of squalene annually in large-scale facilities, presenting a valuable and sustainable alternative for the biotechnological industry and significantly reducing the reliance on non-renewable and endangered sources such as shark liver oil and preventing the annual capture of over 156,661 sharks. Full article

Aurantiochytrium is a well-known long-chain polyunsaturated fatty acids (PUFAs) producer, especially docosahexaenoic acid (DHA). In order to reduce the cost or improve the productivity of DHA, many researchers are focusing on exploring the high-yield strain, reducing production costs, changing culture conditions, and other measures. In this study, DHA production was improved by a two-stage fermentation. In the first stage, efficient and cheap soybean powder was used instead of conventional peptone, and the optimization of fermentation conditions (optimal fermentation conditions: temperature 28.7 °C, salinity 10.7‰, nitrogen source concentration 1.01 g/L, and two-nitrogen ratio of yeast extract to soybean powder 2:1) based on response surface methodology resulted in a 1.68-fold increase in biomass concentration. In the second stage, the addition of 2.5 mM sesamol increased the production of fatty acid and DHA by 93.49% and 98.22%, respectively, as compared to the optimal culture condition with unadded sesamol. Transcriptome analyses revealed that the addition of sesamol resulted in the upregulation of some genes related to fatty acid synthesis and antioxidant enzymes in Aurantiochytrium. This research provides a low-cost and effective culture method for the commercial production of DHA by Aurantiochytrium sp. Full article

This study aimed to evaluate the effects of the microalgae (MIAs) Schizochytrium sp. as a dietary supplement for dairy cows with respect to the yield, composition, and fatty acid profile of milk using a meta-analytical method. The data used in the statistical analyses were obtained from 11 peer-reviewed scientific publications. The effect size was assessed using the weighted mean differences (WMDs) between MIA-supplemented and control treatments. Dry matter intake, milk fat yield, and milk fat content decreased (p < 0.001) in response to the dietary inclusion of Schizochytrium sp. MIAs. However, Schizochytrium sp. MIAs supplementation increased (p = 0.029) milk yield. The dietary inclusion of Schizochytrium sp. MIAs decreased (p < 0.05) the content of the fatty acids (FAs) butyric, caproic, caprylic, capric, undecanoic, lauric, pentadecanoic, palmitic, heptadecanoic, stearic, arachidic, and total saturated FAs, and it resulted in a ω-6/ω-3 ratio in milk. In contrast, Schizochytrium sp. MIAs supplementation increased (p < 0.05) the content of linoleic, conjugated linoleic, eicosapentaenoic, behenic, docosahexaenoic, total monounsaturated FAs, total polyunsaturated FAs, and total omega-3 FAs in milk. The results showed that Schizochytrium sp. MIAs could be used as a dietary supplement to improve the milk yield and fatty acid profile of milk obtained from dairy cows. Full article

A study was conducted to test the inclusion of Aurantiochytrium sp. meal in the rearing of Penaeus vannamei grown in a clear water system and at a suboptimal temperature of 22 °C. The doses tested were 0 (control), 1, 2, 3, and 4% at a stocking density of 100 shrimp/m³. Rearing was carried out with aeration and individual heaters, and seawater temperature was controlled with a chiller. After nine weeks, shrimp were weighed and hemolymph was collected for hemato-immunological tests, and growth performance were calculated. Shrimp raised at 22 °C and fed a 1% of dietary supplementation of Aurantiochytrium sp. meal were superior in immunological parameters. After the dietary assay, a total of 42 shrimps (10.9 ± 0.06 g) per treatment were infected with White Spot Syndrome Virus (WSSV). They were orally infected with 2.6 ± 10⁶ virus particles per g of the animal, maintained at a suboptimal temperature of 22 °C for 108 h, and acclimated to an optimal temperature of 28 °C for an additional 48 h. At 7 days post infection, surviving shrimp were collected for hemato-immunological analysis. Cumulative mortality results showed that shrimp fed diets containing 3% and 4% Aurantiochytrium sp. meal had higher survival than other treatments when challenged with WSSV. Full article

Cellular residues after lipid extraction are the primary waste by-products of microbial polyunsaturated fatty acid production. To mitigate the discharge of cellular residues and reduce the cost of DHA and ARA production, this study examined the utilization of enzymatically hydrolyzed Aurantiochytrium cellular residues (ACRs) and Mortierella alpina cellular residues (MCRs) as nitrogen sources. Results demonstrated that ACRs and MCRs could partially substitute yeast extract (YE) without adverse effects on DHA and ARA fermentation. Moreover, the implementation of a new fermentation medium incorporating ACRs, MCRs, and YE as mixed nitrogen sources resulted in DHA and ARA yields of 17.78 and 5.77 g/L, respectively. These values represented increases of 10.37% and 9.28% compared to traditional cultural methods, while simultaneously reducing the usage of YE by 80% and 60%, respectively. Therefore, the strategy of recycling waste cellular residues presents a novel approach for reducing the costs and environmental impact associated with microbial fermentations. Full article

Aurantiochytrium sp. has received much attention as a potential resource for mass production of omega-3 fatty acids, which contribute to improved growth and reproduction in aquatic animals. In this study, we evaluated the gonadal index changes in zebrafish supplemented with 1–3% Aurantiochytrium sp. crude extract (TE) and the effects of ex vivo environmental Aurantiochytrium sp. on oocytes. 1% TE group showed significant improvement in the gonadal index, and both in vitro incubation and intraperitoneal injection promoted the maturation of zebrafish oocytes. In contrast, the transcriptome revealed 576 genes that were differentially expressed between the 1% TE group and the control group, including 456 up-regulated genes and 120 down-regulated genes. Kyoto Encyclopedia of Genes and Genomes (KEGG) and Gene Ontology (GO) pathway analysis of differentially expressed genes indicated that Aurantiochytrium sp. potentially affects pathways such as lipid metabolism, immune regulation, and oocyte development in zebrafish. The results of this study enriched the knowledge of Aurantiochytrium sp. in regulating gonadal development in zebrafish and provided a theoretical basis for its application in aquaculture. Full article

Microalgae are a promising feedstock for bioethanol production, essentially due to their high growth rates and absence of lignin. Hydrolysis—where the monosaccharides are released for further fermentation—is considered a critical step, and its optimization is advised for each raw material. The present study focuses on the thermal acid hydrolysis (with sulfuric acid) of Aurantiochytrium sp. through a response surface methodology (RSM), studying the effect of acid concentration, hydrolysis time and biomass/acid ratio on both sugar concentration of the hydrolysate and biomass conversion yield. Preliminary studies allowed to establish the range of the variables to be optimized. The obtained models predicted a maximum sugar concentration (18.05 g/L; R² = 0.990) after 90 min of hydrolysis, using 15% (w/v) biomass/acid ratio and sulfuric acid at 3.5% (v/v), whereas the maximum conversion yield (12.86 g/100 g; R² = 0.876) was obtained using 9.3% (w/v) biomass/acid ratio, maintaining the other parameters. Model outputs indicate that the biomass/acid ratio and time are the most influential parameters on the sugar concentration and yield models, respectively. The study allowed to obtain a predictive model that is very well adjusted to the experimental data to find the best saccharification conditions for the Aurantiochytrium sp. microalgae. Full article

Media supplementation has proven to be an effective technique for improving byproduct yield during microbial fermentation. This study explored the impact of different concentrations of bioactive compounds, namely alpha-tocopherol, mannitol, melatonin, sesamol, ascorbic acid, and biotin, on the Aurantiochytrium sp. TWZ-97 culture. Our investigation revealed that alpha-tocopherol was the most effective compound in reducing the reactive oxygen species (ROS) burden, both directly and indirectly. Adding 0.7 g/L of alpha-tocopherol led to an 18% improvement in biomass, from 6.29 g/L to 7.42 g/L. Moreover, the squalene concentration increased from 129.8 mg/L to 240.2 mg/L, indicating an 85% improvement, while the squalene yield increased by 63.2%, from 19.82 mg/g to 32.4 mg/g. Additionally, our comparative transcriptomics analysis suggested that several genes involved in glycolysis, pentose phosphate pathway, TCA cycle, and MVA pathway were overexpressed following alpha-tocopherol supplementation. The alpha-tocopherol supplementation also lowered ROS levels by binding directly to ROS generated in the fermentation medium and indirectly by stimulating genes that encode antioxidative enzymes, thereby decreasing the ROS burden. Our findings suggest that alpha-tocopherol supplementation can be an effective method for improving squalene production in Aurantiochytrium sp. TWZ-97 culture. Full article

The use of chitosan as a flocculant has become a topic of interest over the years due to its positively charged polymer and biodegradable and non-toxic properties. However, most studies only focus on microalgae and wastewater treatment. This study provides crucial insight into the potential of using chitosan as an organic flocculant to harvest lipids and docosahexaenoic acid (DHA-rich Aurantiochytrium sp. SW1 cells by examining the correlation of flocculation parameters (chitosan concentration, molecular weight, medium pH, culture age, and cell density) toward the flocculation efficiency and zeta potential of the cells. A strong correlation between the pH and harvesting efficiency was observed as the pH increased from 3, with the optimal flocculation efficiency of >95% achieved at a chitosan concentration of 0.5 g/L at pH 6 where the zeta potential was almost zero (3.26 mV). The culture age and chitosan molecular weight have no effect on the flocculation efficiency but increasing the cell density decreases the flocculation efficiency. This is the first study to reveal the potential of chitosan to be used as a harvesting alternative for thraustochytrid cells. Full article

Precursor regulation has been an effective strategy to improve carotenoid production and the availability of novel precursor synthases facilitates engineering improvements. In this work, the putative geranylgeranyl pyrophosphate synthase encoding gene (AlGGPPS) and isopentenyl pyrophosphate isomerase encoding gene (AlIDI) from Aurantiochytrium limacinum MYA-1381 were isolated. We applied the excavated AlGGPPS and AlIDI to the de novo β-carotene biosynthetic pathway in Escherichia coli for functional identification and engineering application. Results showed that the two novel genes both functioned in the synthesis of β-carotene. Furthermore, AlGGPPS and AlIDI performed better than the original or endogenous one, with 39.7% and 80.9% increases in β-carotene production, respectively. Due to the coordinated expression of the 2 functional genes, β-carotene content of the modified carotenoid-producing E. coli accumulated a 2.99-fold yield of the initial EBIY strain in 12 h, reaching 10.99 mg/L in flask culture. This study helped to broaden current understanding of the carotenoid biosynthetic pathway in Aurantiochytrium and provided novel functional elements for carotenoid engineering improvements. Full article

Thraustochytrids are unicellular marine heterotrophic protists, which have recently shown a promising ability to produce omega-3 fatty acids from lignocellulosic hydrolysates and wastewaters. Here we studied the biorefinery potential of the dilute acid-pretreated marine macroalgae (Enteromorpha) in comparison with glucose via fermentation using a previously isolated thraustochytrid strain (Aurantiochytrium limacinum PKU#Mn4). The total reducing sugars in the Enteromorpha hydrolysate accounted for 43.93% of the dry cell weight (DCW). The strain was capable of producing the highest DCW (4.32 ± 0.09 g/L) and total fatty acids (TFA) content (0.65 ± 0.03 g/L) in the medium containing 100 g/L of hydrolysate. The maximum TFA yields of 0.164 ± 0.160 g/g DCW and 0.196 ± 0.010 g/g DCW were achieved at 80 g/L of hydrolysate and 40 g/L of glucose in the fermentation medium, respectively. Compositional analysis of TFA revealed the production of equivalent fractions (% TFA) of saturated and polyunsaturated fatty acids in hydrolysate or glucose medium. Furthermore, the strain yielded a much higher fraction (2.61–3.22%) of eicosapentaenoic acid (C20:5n-3) in the hydrolysate medium than that (0.25–0.49%) in the glucose medium. Overall, our findings suggest that Enteromorpha hydrolysate can be a potential natural substrate in the fermentative production of high-value fatty acids by thraustochytrids. Full article

Thraustochytrids are aquatic unicellular protists organisms that represent an important reservoir of a wide range of bioactive compounds, such as essential polyunsaturated fatty acids (PUFAs) such as arachidonic acid (ARA), docosahexaenoic acid (DHA), eicosapentaenoic acid (EPA), which are involved in the regulation of the immune system. In this study, we explore the use of co-cultures of Aurantiochytrium sp. and bacteria as a biotechnological tool capable of stimulating PUFA bioaccumulation. In particular, the co-culture of lactic acid bacteria and the protist Aurantiochytrium sp. T66 induce PUFA bioaccumulation, and the lipid profile was evaluated in cultures at different inoculation times, with two different strains of lactic acid bacteria capable of producing the tryptophan dependent auxins, and one strain of Azospirillum sp., as a reference for auxin production. Our results showed that the Lentilactobacillus kefiri K6.10 strain inoculated at 72 h gives the best PUFA content (30.89 mg g⁻¹ biomass) measured at 144 h of culture, three times higher than the control (8.87 mg g⁻¹ biomass). Co-culture can lead to the generation of complex biomasses with higher added value for developing aquafeed supplements. Full article

The study of heterotrophic protists in the ocean is still in its early stages, compared to other microorganisms such as bacteria, archaea, and photoautotrophic protists. Labyrinthulomycetes protists (LP) are a type of unicellular protists that are widely distributed in global waters and have the potential to produce high-value products. In this study, the abundance, diversity, and community structure of LP in the coastal zone of Hainan Island in the South China Sea were investigated through quantitative PCR and high-throughput sequencing. The results showed that LP abundance varied by location and depth, with the highest levels (37.3 × 10³ copies/L) found in the middle layer offshore and the lowest (0.386 × 10³ copies/L) in the bottom layer offshore. The middle layer (chlorophyll maximum layer) had higher LP abundance both inshore and offshore than the surface and bottom layers. Interestingly, the highest LP richness and diversity was found in the inshore bottom. There was a significant difference in LP abundance between the offshore surface and bottom layers. The LP community was dominated by the genus Aplanochytrium, and four different ecotypes were identified. Additionally, the genus Aurantiochytrium had different cooperative and competitive strategies with bacteria in different habitats. This study sheds light on the abundance and community structure of LP in the coastal zone of Hainan Island, explores the potential interactions between LP and bacterial populations, and raises questions about the potential differentiation of LP ecotypes. Full article

An 8-week feeding trial investigated the effect of Fishmeal (FM) replacement by soybean meal (SBM) and poultry by-product meal (PBM) in diets supplemented with DL-Met, MET-MET (AQUAVI^®), Bacillus amyloliquefaciens CECT 5940 (ECOBIOL^®) and their combinations on growth performance and health of juvenile Litopenaeus vannamei. A total of six experimental diets were formulated according to L. vannamei nutritional requirements. A total of 480 shrimp (0.30 ± 0.04 g) were randomly distributed into 24 tanks (4 repetitions/each diet, 20 shrimp/tank). Shrimp were fed with control diet (CD; 200 g/Kg fishmeal) and five diets with 50% FM replacement supplemented with different methionine sources, probiotic (B. amyloliquefaciens CECT 5940) and their combinations: D1 (0.13% DL-MET), D2 (0.06% MET-MET), D3 (0.19% MET-MET), D4 (0.13% DL-MET plus 0.10% B. amyloliquefaciens CECT 5940 and D5 (0.06% MET-MET plus 0.10% B. amyloliquefaciens CECT 5940). Shrimp fed D3 and D5 had significantly higher final, weekly weight gain, and final biomass compared to shrimp fed CD (p < 0.05). Shrimp fed D2 to D5 increased the hepatopancreas epithelial cell height (p < 0.05). Digestive enzymatic activities were significantly increased in shrimp hepatopancreas’ fed D3 (p < 0.05). Meanwhile, shrimp fed D1 had significant downregulation of immune-related genes (p < 0.05). Moreover, shrimp fed D3 and D5 increased the abundance of beneficial prokaryotic microorganisms such as Pseudoalteromonas and Demequina related to carbohydrate metabolism and immune stimulation. Also, shrimp fed D3 and D5 increased the abundance of beneficial eukaryotic microorganism as Aurantiochytrium and Aplanochytrium were related to eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) production which plays a role in growth promoting or boosting the immunity of aquatic organisms. Therefore, fishmeal could be partially substituted up to 50% by SBM and PBM in diets supplemented with 0.19% MET-MET (AQUAVI^®) or 0.06% MET-MET (AQUAVI^®) plus 0.10% B. amyloliquefaciens CECT 5940 (ECOBIOL^®) and improve the productive performance, health, and immunity of white shrimp. Further research is necessary to investigate synergistic effects of amino acids and probiotics in farmed shrimp diets, as well as to evaluate how SBM and PBM influence the fatty acid composition of reduced fishmeal diets and shrimp muscle quality. Nevertheless, this information could be interesting to develop low fishmeal feeds for aquaculture without affecting the growth and welfare of aquatic organisms. Full article