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1.
Sci Rep ; 14(1): 14346, 2024 06 21.
Article de Anglais | MEDLINE | ID: mdl-38906947

RÉSUMÉ

This study investigated the first-ever reported use of freshwater Nannochloropsis for the bioremediation of dairy processing side streams and co-generation of valuable products, such as ß-galactosidase enzyme. In this study, N. limnetica was found to grow rapidly on both autoclaved and non-autoclaved whey-powder media (referred to dairy processing by-product or DPBP) without the need of salinity adjustment or nutrient additions, achieving a biomass concentration of 1.05-1.36 g L-1 after 8 days. The species secreted extracellular ß-galactosidase (up to 40.84 ± 0.23 U L-1) in order to hydrolyse lactose in DPBP media into monosaccharides prior to absorption into biomass, demonstrating a mixotrophic pathway for lactose assimilation. The species was highly effective as a bioremediation agent, being able to remove > 80% of total nitrogen and phosphate in the DPBP medium within two days across all cultures. Population analysis using flow cytometry and multi-channel/multi-staining methods revealed that the culture grown on non-autoclaved medium contained a high initial bacterial load, comprising both contaminating bacteria in the medium and phycosphere bacteria associated with the microalgae. In both autoclaved and non-autoclaved DPBP media, Nannochloropsis cells were able to establish a stable microalgae-bacteria interaction, suppressing bacterial takeover and emerging as dominant population (53-80% of total cells) in the cultures. The extent of microalgal dominance, however, was less prominent in the non-autoclaved media. High initial bacterial loads in these cultures had mixed effects on microalgal performance, promoting ß-galactosidase synthesis on the one hand while competing for nutrients and retarding microalgal growth on the other. These results alluded to the need of effective pre-treatment step to manage bacterial population in microalgal cultures on DPBP. Overall, N. limnetica cultures displayed competitive ß-galactosidase productivity and propensity for efficient nutrient removal on DPBP medium, demonstrating their promising nature for use in the valorisation of dairy side streams.


Sujet(s)
Microalgues , Lactosérum , beta-Galactosidase , beta-Galactosidase/métabolisme , Microalgues/métabolisme , Microalgues/enzymologie , Lactosérum/métabolisme , Lactose/métabolisme , Straménopiles/enzymologie , Straménopiles/métabolisme , Eau douce/microbiologie , Dépollution biologique de l'environnement , Biomasse , Azote/métabolisme
2.
Int J Mol Sci ; 25(12)2024 Jun 20.
Article de Anglais | MEDLINE | ID: mdl-38928505

RÉSUMÉ

Nannochloropsis gaditana, a microalga known for its photosynthetic efficiency, serves as a cell factory, producing valuable biomolecules such as proteins, lipids, and pigments. These components make it an ideal candidate for biofuel production and pharmaceutical applications. In this study, we genetically engineered N. gaditana to overexpress the enzyme fructose-1,6-bisphosphatase (cyFBPase) using the Hsp promoter, aiming to enhance sugar metabolism and biomass accumulation. The modified algal strain, termed NgFBP, exhibited a 1.34-fold increase in cyFBPase activity under photoautotrophic conditions. This modification led to a doubling of biomass production and an increase in eicosapentaenoic acid (EPA) content in fatty acids to 20.78-23.08%. Additionally, the genetic alteration activated the pathways related to glycine, protoporphyrin, thioglucosides, pantothenic acid, CoA, and glycerophospholipids. This shift in carbon allocation towards chloroplast development significantly enhanced photosynthesis and growth. The outcomes of this study not only improve our understanding of photosynthesis and carbon allocation in N. gaditana but also suggest new biotechnological methods to optimize biomass yield and compound production in microalgae.


Sujet(s)
Biomasse , Fructose-1,6-diphosphatase , Métabolomique , Microalgues , Photosynthèse , Straménopiles , Fructose-1,6-diphosphatase/métabolisme , Fructose-1,6-diphosphatase/génétique , Straménopiles/génétique , Straménopiles/métabolisme , Straménopiles/croissance et développement , Straménopiles/enzymologie , Microalgues/métabolisme , Microalgues/génétique , Microalgues/croissance et développement , Microalgues/enzymologie , Métabolomique/méthodes , Cytosol/métabolisme
3.
Nat Commun ; 15(1): 5456, 2024 Jun 27.
Article de Anglais | MEDLINE | ID: mdl-38937455

RÉSUMÉ

Stramenopile algae contribute significantly to global primary productivity, and one class, Eustigmatophyceae, is increasingly studied for applications in high-value lipid production. Yet much about their basic biology remains unknown, including the nature of an enigmatic, pigmented globule found in vegetative cells. Here, we present an in-depth examination of this "red body," focusing on Nannochloropsis oceanica. During the cell cycle, the red body forms adjacent to the plastid, but unexpectedly it is secreted and released with the autosporangial wall following cell division. Shed red bodies contain antioxidant ketocarotenoids, and overexpression of a beta-carotene ketolase results in enlarged red bodies. Infrared spectroscopy indicates long-chain, aliphatic lipids in shed red bodies and cell walls, and UHPLC-HRMS detects a C32 alkyl diol, a potential precursor of algaenan, a recalcitrant cell wall polymer. We propose that the red body transports algaenan precursors from plastid to apoplast to be incorporated into daughter cell walls.


Sujet(s)
Paroi cellulaire , Plastes , Straménopiles , Paroi cellulaire/métabolisme , Straménopiles/métabolisme , Plastes/métabolisme
4.
Bioresour Technol ; 406: 131024, 2024 Aug.
Article de Anglais | MEDLINE | ID: mdl-38914238

RÉSUMÉ

The rising prices of fishery derivatives limits their use in aquafeeds. Therefore, other alternatives are used to replace those ingredients. Among them, microalgae are of great interest both as an ingredient and as a potential stabilising agent against lipid oxidation. This study evaluates on the use of Nannochloropsis gaditana to prevent lipid oxidation in a set of 12 aquafeeds over 540 days of storage. Aquafeeds were formulated with/without 15 % N. gaditana combined with two antioxidants -butylhydroxytoluene (25-150 mg·kg-1) or vitamin E (500-3000 mg·kg-1). The effect of i) storage period, ii) presence of microalgae and iii) antioxidant addition on lipid oxidation was assessed. Results showed higher fatty acid degradation in diets lacking microalgae. The microalgae supplemented diets is enough for preserving feeds presenting the highest antioxidant effect at the end, without significant differences with the microalgae-supplemented feeds and those including antioxidants after 540 days of storage.


Sujet(s)
Aliment pour animaux , Antioxydants , Aquaculture , Microalgues , Microalgues/métabolisme , Antioxydants/pharmacologie , Aquaculture/méthodes , Straménopiles/métabolisme , Oxydoréduction , Stockage des aliments/méthodes , Acides gras , Peroxydation lipidique/effets des médicaments et des substances chimiques
5.
Bioresour Technol ; 403: 130891, 2024 Jul.
Article de Anglais | MEDLINE | ID: mdl-38788808

RÉSUMÉ

To reduce the cost of docosahexaenoic acid (DHA) production from Schizochytrium sp., the waste Pichia pastoris was successfully used as an alternative nitrogen source to achieve high-density cultivation during the cell growth phase. However, due to the high oxygen consumption feature when implementing high-density cultivation, the control of both the nitrogen source and dissolved oxygen concentration (DO) at each sufficient level was impossible; thus, two realistic control strategies, including "DO sufficiency-nitrogen limitation" and "DO limitation-nitrogen sufficiency", were proposed. When using the strategy of "DO sufficiency-nitrogen limitation", the lowest maintenance coefficient of glucose (12.3 mg/g/h vs. 17.0 mg/g/h) and the highest activities of related enzymes in DHA biosynthetic routes were simultaneously obtained; thus, a maximum DHA concentration of 12.8 ± 1.2 g/L was achieved, which was 1.58-fold greater than that of the control group. Overall, two-stage feeding control for alternative nitrogen sources is an efficient strategy to industrial DHA fermentation.


Sujet(s)
Acide docosahexaénoïque , Azote , Straménopiles , Acide docosahexaénoïque/métabolisme , Acide docosahexaénoïque/biosynthèse , Azote/métabolisme , Straménopiles/métabolisme , Fermentation , Oxygène/métabolisme , Glucose/métabolisme , Saccharomycetales/métabolisme
6.
Bioresour Technol ; 402: 130829, 2024 Jun.
Article de Anglais | MEDLINE | ID: mdl-38734261

RÉSUMÉ

Most marine microalgae are typically cultivated in coastal areas due to challenges in inland cultivation. In this 185 days experiment, Nannochloropsis oceanica was semi-continuously cultivated inland using different photobioreactors (PBRs). The newly designed 700-liter (L) PBR exhibited tolerance to seasonal changes compared to the 150-L PBRs. The innovative in-situ oxygen release rate (ORR) measurement method results indicated that ORR was influenced by light intensity and temperature. The optimal temperature range for N. oceanica growth was 14-25 â„ƒ, demonstrated cold tolerance and lipid accumulation at low temperatures. The maximum lipid content in 700-L and 150-L PBRs was 29 % and 28 %, respectively. Based on the average biomass productivity, the price of N. oceanica was $11.89 kg-1 (or $3.35 kg-1 based on maximum biomass productivity), which is cheaper than the current market price of $20.19 kg-1. From results, smaller PBRs at the same hydro electricity price are more cost-effective.


Sujet(s)
Biomasse , Microalgues , Photobioréacteurs , Straménopiles , Microalgues/croissance et développement , Microalgues/métabolisme , Straménopiles/croissance et développement , Straménopiles/métabolisme , Température , Oxygène , Lumière
7.
Plant Physiol Biochem ; 211: 108729, 2024 Jun.
Article de Anglais | MEDLINE | ID: mdl-38754177

RÉSUMÉ

Microalgae, recognized as sustainable and eco-friendly photosynthetic microorganisms, play a pivotal role in converting CO2 into value-added products. Among these, Nannochloropsis salina (Microchloropsis salina) stands out, particularly for its ability to produce eicosapentaenoic acid (EPA), a crucial omega-3 fatty acid with significant health benefits such as anti-inflammatory properties and cardiovascular health promotion. This study focused on optimizing the cultivation conditions of Nannochloropsis salina to maximize EPA production. We thoroughly investigated the effects of varying temperatures and nitrogen (NaNO3) concentrations on biomass, total lipid content, and EPA proportions. We successfully identified optimal conditions at an initial NaNO3 concentration of 1.28 g.L-1 and a temperature of 21 °C. This condition was further validated by response surface methodology, which resulted in the highest EPA productivity reported in batch systems (14.4 mg.L-1.day-1). Quantitative real-time PCR and transcriptomic analysis also demonstrated a positive correlation between specific gene expressions and enhanced EPA production. Through a comprehensive lipid analysis and photosynthetic pigment analysis, we deduced that the production of EPA in Nannochloropsis salina seemed to be produced by the remodeling of chloroplast membrane lipids. These findings provide crucial insights into how temperature and nutrient availability influence fatty acid composition in N. salina, offering valuable guidance for developing strategies to improve EPA production in various microalgae species.


Sujet(s)
Acide eicosapentanoïque , Microalgues , Azote , Photosynthèse , Straménopiles , Température , Acide eicosapentanoïque/métabolisme , Acide eicosapentanoïque/biosynthèse , Azote/métabolisme , Microalgues/métabolisme , Straménopiles/métabolisme , Straménopiles/génétique , Biomasse
8.
Bioprocess Biosyst Eng ; 47(6): 863-875, 2024 Jun.
Article de Anglais | MEDLINE | ID: mdl-38687387

RÉSUMÉ

Eicosapentaenoic acid (EPA) is a vital É·-3 polyunsaturated fatty acid (PUFA) for human body with various physiological functions. In this study, we proposed an adaptive evolutionary strategy based on high-temperature and high-oxygen two-factor stress to increase the EPA production capacity of Schizochytrium. High-temperature stress was used to increase EPA yield, and high oxygen was implemented to continuously stimulate cell growth and lipid accumulation. The biomass and EPA production of ALE-D50 reached 35.33 g/L and 1.54 g/L, which were 43.85% and 71.11% higher than that of the original strain, respectively. Lower in vivo reactive oxygen species levels indicated that the evolved strain possessed stronger antioxidant activity. Liquid chromatography-mass spectrometry metabolomics showed that enhanced glucose consumption and glycolysis metabolism, as well as a weakened tricarboxylic acid cycle and reduced amino acid metabolic tributaries in the evolved strain, might be associated with increased growth and EPA synthesis. Finally, the lipid production and EPA production in a fed-batch fermentation were further increased to 48.93 g/L and 3.55 g/L, improving by 54.30% and 90.86%, respectively. This study provides a novel pathway for promoting EPA biosynthesis in Schizochytrium.


Sujet(s)
Acide eicosapentanoïque , Métabolomique , Straménopiles , Acide eicosapentanoïque/biosynthèse , Acide eicosapentanoïque/métabolisme , Straménopiles/métabolisme , Straménopiles/croissance et développement , Stress physiologique , Adaptation physiologique
9.
Biotechnol Adv ; 73: 108354, 2024.
Article de Anglais | MEDLINE | ID: mdl-38588906

RÉSUMÉ

Thraustochytrids are marine microorganisms known for their fast growth and ability to store lipids, making them useful for producing polyunsaturated fatty acids (PUFAs), biodiesel, squalene, and carotenoids. However, the high cost of production, mainly due to expensive fermentation components, limits their wider use. A significant challenge in this context is the need to balance production costs with the value of the end products. This review focuses on integrating the efficient utilization of waste with Thraustochytrids fermentation, including the economic substitution of carbon sources, nitrogen sources, and fermentation water. This approach aligns with the 3Rs principles (reduction, recycling, and reuse). Furthermore, it emphasizes the role of Thraustochytrids in converting waste into lipid chemicals and promoting sustainable circular production models. The aim of this review is to emphasize the value of Thraustochytrids in converting waste into treasure, providing precise cost reduction strategies for future commercial production.


Sujet(s)
Conservation des ressources naturelles , Fermentation , Straménopiles , Déchets , Biocarburants , Biotechnologie/économie , Biotechnologie/méthodes , Carbone/métabolisme , Acides gras insaturés/métabolisme , Lipides/biosynthèse , Lipides/composition chimique , Straménopiles/métabolisme
10.
Bioresour Technol ; 398: 130537, 2024 Apr.
Article de Anglais | MEDLINE | ID: mdl-38452955

RÉSUMÉ

Antioxidant addition is an effective strategy to achieve docosahexaenoic acid (DHA) overproduction in oleaginous microorganisms. Nevertheless, antioxidants like phenolic compounds sometimes exert pro-oxidant activity. In this work, effects of proanthocyanidins (PAs) on fermentation performance and oxidative stress in Schizochytrium sp. were investigated. Low PAs addition (5 mg/L) reduced reactive oxygen species and enhanced lipogenic enzymes activities and NADPH, resulting in significant increase in lipid (20.3 g/L) by 33.6 % and DHA yield (9.8 g/L) by 53.4 %. In contrast, high PAs addition (500 mg/L) exerted pro-oxidant effects, aggravated oxidative damage and lipid peroxidation, leading to sharp decrease in biomass (21.3 g/L) by 35.1 %, lipid (8.2 g/L) by 46.0 %, and DHA (2.9 g/L) by 54.8 %. Therefore, the antioxidant concentration is especially crucial in DHA production. This study is the first to report concentration-dependant dual roles of PAs in oxidative stress and DHA production in Schizochytrium sp., providing new insights into microbial DHA production.


Sujet(s)
Proanthocyanidines , Straménopiles , Antioxydants/métabolisme , Acide docosahexaénoïque , Proanthocyanidines/pharmacologie , Espèces réactives de l'oxygène , Straménopiles/métabolisme , Stress oxydatif , Fermentation
11.
Biotechnol Lett ; 46(3): 373-383, 2024 Jun.
Article de Anglais | MEDLINE | ID: mdl-38493279

RÉSUMÉ

OBJECTIVE: Currently, there is lack of a consistent and highly enriched source for docosapentaenoic acid (n-3 DPA, C22:5), and this work report the isolation of microorganism that naturally produces n-3 DPA. RESULTS: In this work, we screened microorganisms in our culture collections with the goal to isolate a strain with high levels of n-3 DPA. We isolated a strain of Sphaeroforma arctica that produces up to 11% n-3 DPA in total fatty acid and has a high n-3 DPA to DHA/EPA ratio. The cell growth of the isolated strain was characterized using microscopy imaging and flow cytometer technologies to confirm the coenocytic pattern of cell divisions previously described in S. arctica. Our novel isolate of S. arctica grew more robustly and produced significantly more n-3 DPA compared to previously isolated and described strains indicating the uniqueness of the discovered strain. CONCLUSION: Overall, this work reports a first isolate n-3 DPA producing microorganism and establishes the foundation for future strain improvement and elucidation of the physiological function of this LC-PUFA for human nutrition and health.


Sujet(s)
Acides gras insaturés , Acides gras insaturés/métabolisme , Straménopiles/métabolisme , Straménopiles/isolement et purification
12.
Biosci Biotechnol Biochem ; 88(6): 696-704, 2024 May 22.
Article de Anglais | MEDLINE | ID: mdl-38520162

RÉSUMÉ

We focused on the production of docosahexaenoic acid (DHA)-containing microbial lipids by Aurantiochytrium sp. using of defatted soybean (DS) as a nitrogen source. Defatted soybean is a plant biomass that could provide a sustainable supply at a low cost. Results showed that Aurantiochytrium sp. could not directly assimilate the DS as a nitrogen source but could grow well in a medium containing DS fermented with rice malt. When cultivated in a fermented DS (FDS) medium, Aurantiochytrium sp. showed vigorous growth with the addition of sufficient sulfate and chloride ions as inorganic nutrients without seawater salt. A novel isolated Aurantiochytrium sp. 6-2 showed 15.8 ± 3.4 g/L DHA productivity (in 54.8 ± 12.1 g/L total fatty acid production) in 1 L of the FDS medium. Therefore, DHA produced by Aurantiochytrium sp. using FDS enables a stable and sustainable DHA supply and could be an alternative source of natural DHA derived from fish oil.


Sujet(s)
Aliment pour animaux , Acide docosahexaénoïque , Fermentation , Glycine max , Azote , Straménopiles , Acide docosahexaénoïque/biosynthèse , Acide docosahexaénoïque/métabolisme , Glycine max/métabolisme , Glycine max/croissance et développement , Azote/métabolisme , Straménopiles/métabolisme , Straménopiles/croissance et développement , Aliment pour animaux/analyse , Animaux , Poissons/métabolisme , Biomasse , Milieux de culture/composition chimique
13.
Biotechnol Bioeng ; 121(6): 1986-2001, 2024 Jun.
Article de Anglais | MEDLINE | ID: mdl-38500406

RÉSUMÉ

Marine thraustochytrids produce metabolically important lipids such as the long-chain omega-3 polyunsaturated fatty acids, carotenoids, and sterols. The growth and lipid production in thraustochytrids depends on the composition of the culture medium that often contains yeast extract as a source of amino acids. This work discusses the effects of individual amino acids provided in the culture medium as the only source of nitrogen, on the production of biomass and lipids by the thraustochytrid Thraustochytrium sp. RT2316-16. A reconstructed metabolic network based on the annotated genome of RT2316-16 in combination with flux balance analysis was used to explain the observed growth and consumption of the nutrients. The culture kinetic parameters estimated from the experimental data were used to constrain the flux via the nutrient consumption rates and the specific growth rate of the triacylglycerol-free biomass in the genome-scale metabolic model (GEM) to predict the specific rate of ATP production for cell maintenance. A relationship was identified between the specific rate of ATP production for maintenance and the specific rate of glucose consumption. The GEM and the derived relationship for the production of ATP for maintenance were used in linear optimization problems, to successfully predict the specific growth rate of RT2316-16 in different experimental conditions.


Sujet(s)
Modèles biologiques , Straménopiles , Straménopiles/métabolisme , Straménopiles/génétique , Milieux de culture/composition chimique , Milieux de culture/métabolisme , Voies et réseaux métaboliques/génétique , Acides aminés/métabolisme , Biomasse , Métabolisme lipidique , Nutriments/métabolisme , Adénosine triphosphate/métabolisme
14.
BMC Genomics ; 25(1): 141, 2024 Feb 05.
Article de Anglais | MEDLINE | ID: mdl-38311722

RÉSUMÉ

BACKGROUND: Schizochytrium limacinum holds significant value utilized in the industrial-scale synthesis of natural DHA. Nitrogen-limited treatment can effectively increase the content of fatty acids and DHA, but there is currently no research on chromatin accessibility during the process of transcript regulation. The objective of this research was to delve into the workings of fatty acid production in S. limacinum by examining the accessibility of promoters and profiling gene expressions. RESULTS: Results showed that differentially accessible chromatin regions (DARs)-associated genes were enriched in fatty acid metabolism, signal transduction mechanisms, and energy production. By identifying and annotating DARs-associated motifs, the study obtained 54 target transcription factor classes, including BPC, RAMOSA1, SPI1, MYC, and MYB families. Transcriptomics results revealed that several differentially expressed genes (DEGs), including SlFAD2, SlALDH, SlCAS1, SlNSDHL, and SlDGKI, are directly related to the biosynthesis of fatty acids, meanwhile, SlRPS6KA, SlCAMK1, SlMYB3R1, and SlMYB3R5 serve as transcription factors that could potentially influence the regulation of fatty acid production. In the integration analysis of DARs and ATAC-seq, 13 genes were identified, which were shared by both DEGs and DARs-associated genes, including SlCAKM, SlRP2, SlSHOC2, SlTN, SlSGK2, SlHMP, SlOGT, SlclpB, and SlDNAAF3. CONCLUSIONS: SlCAKM may act as a negative regulator of fatty acid and DHA synthesis, while SlSGK2 may act as a positive regulator, which requires further study in the future. These insights enhance our comprehension of the processes underlying fatty acid and DHA production in S. limacinum. They also supply a foundational theoretical framework and practical assistance for the development of strains rich in fatty acids and DHA.


Sujet(s)
Séquençage après immunoprécipitation de la chromatine , Straménopiles , Humains , RNA-Seq , Azote/métabolisme , Acides gras/métabolisme , Chromatine/métabolisme , Acide docosahexaénoïque , Straménopiles/génétique , Straménopiles/métabolisme
15.
Sci Total Environ ; 912: 169217, 2024 Feb 20.
Article de Anglais | MEDLINE | ID: mdl-38081429

RÉSUMÉ

Aurantiochytrium sp. belongs to marine heterotrophic single-cell protist, which is an important decomposer in marine ecosystem. Aurantiochytrium sp. has gained notoriety because of its ability to accumulate high-value docosahexaenoic acid (DHA), but the key factors of DHA synthesis were unclear at present. In this study, Atmospheric and Room Temperature Plasma technology was applied to the mutagenic breeding of Aurantiochytrium sp., and transcriptomics and proteomics were adopted to analyze the DHA-biosynthesis mechanism. According to the growth and DHA accumulation profiles, the mutant strain Aurantiochytrium sp. R2A35 was selected. The DHA content in total lipids was greatly improved from 49.39 % of the wild strain R2 to 63.69 % of the mutant strain. Moreover, the DHA content in the biomass of Aurantiochytrium sp. R2A35 as 39.72 % was the highest DHA productivity reported so far. The differentially expressed genes distinguished from transcriptome and the TMT-identified differential proteins distinguished from proteome confirmed that the expression of acetyl-CoA carboxylase and ketoacyl reductase was up-regulated by 4.78-fold and 6.95-fold, respectively and the fatty acid synthase was concurrently down-regulated by 2.79-fold, so that more precursor was transported to the polyketide synthase pathway, thereby increasing the DHA yield in Aurantiochytrium sp. R2A35. This research would provide reference for the DHA metabolism process and contribute to the understanding of the decomposer - Aurantiochytrium sp. in marine ecosystems.


Sujet(s)
Acide docosahexaénoïque , Straménopiles , Acide docosahexaénoïque/métabolisme , Écosystème , Température , Multi-omique , Straménopiles/métabolisme , Mutagenèse
16.
Free Radic Biol Med ; 210: 246-257, 2024 01.
Article de Anglais | MEDLINE | ID: mdl-38042223

RÉSUMÉ

Schizochytrium sp. is an important industrial strain for commercial production of docosahexaenoic acid (DHA), which plays essential physiological roles in infant development and human health. The regulatory network for DHA biosynthesis and lipid accumulation in Schizochytrium remains poorly understood. FabR (fatty acid biosynthesis repressor), a basic leucine zipper (bZIP) transcription factor, was transcriptionally downregulated under low-nitrogen condition. Deletion of fabR gene (mutant ΔfabR) increased production of total lipids and DHA by 30.1% and 46.5%, respectively. ΔfabR displayed H2O2 stress resistance higher than that of parental strain or complementation strain CfabR. FabR bound specifically to 7-bp pseudo-palindromic sequence 5'-ATTSAAT-3' in upstream regions and repressed transcription of fatty acid biosynthesis genes (acl, fas, pfa) and antioxidant defense genes (cat, sod1, sod2, gpx). DNA binding activity of FabR was regulated in a redox-dependent manner. Under oxidative condition, FabR forms intermolecular disulfide bonds between two Cys46 residues of dimers; its DNA binding activity is thereby lost, and the transcription of its target genes is enhanced through derepression. Our findings clarify the redox-dependent mechanism that modulates FabR activity governing lipid and DHA biosynthesis and H2O2 stress response in Schizochytrium.


Sujet(s)
Acide docosahexaénoïque , Straménopiles , Enfant , Humains , Acide docosahexaénoïque/génétique , Acide docosahexaénoïque/métabolisme , Peroxyde d'hydrogène , Facteurs de transcription à motif basique et à glissière à leucines/métabolisme , Straménopiles/métabolisme , Oxydoréduction , ADN/métabolisme
17.
Bioresour Technol ; 394: 130205, 2024 Feb.
Article de Anglais | MEDLINE | ID: mdl-38104661

RÉSUMÉ

Tribonema minus, a promising filamentous oleaginous microalga, was cultured under different nutrient concentrations and different culture modes (fed-batch culture, two-step culture) to study the method of rapid regulation of its lipid metabolism. In contrast to many other oleaginous microalgae, T. minus did not show that nitrogen stress promoted lipid accumulation; however, sulfur deficiency promoted rapid lipid accumulation with a maximum lipid content of 54% of dry weight. Increasing the MgSO4 concentration significantly increased nitrogen uptake and biomass (10.09 g/L). Lipid productivity was significantly increased by the two-step culture using a medium with a high concentration of MgSO4 in the first step and a sulfur-free medium in the second step. In addition, it was found that the lipid content of T. minus was negatively correlated with the intracellular sulfur content when the intracellular sulfur content was below 0.6%. This study provides a new approach for industrial lipid production in T. minus.


Sujet(s)
Microalgues , Straménopiles , Straménopiles/métabolisme , Microalgues/métabolisme , Biomasse , Azote/métabolisme , Lipides
18.
Biochim Biophys Acta Mol Cell Biol Lipids ; 1868(11): 159381, 2023 11.
Article de Anglais | MEDLINE | ID: mdl-37625782

RÉSUMÉ

Aurantiochytrium sp., a marine thraustochytrid possesses a remarkable ability to produce lipid rich in polyunsaturated fatty acids (PUFAs), such as docosahexaenoic acid (DHA). Although gene regulation underlying lipid biosynthesis has been previously reported, proteomic analysis is still limited. In this study, high DHA accumulating strain Aurantiochytrium sp. SW1 has been used as a study model to elucidate the alteration in proteome profile under different cultivation phases i.e. growth, nitrogen-limitation and lipid accumulation. Of the total of 5146 identified proteins, 852 proteins were differentially expressed proteins (DEPs). The largest number of DEPs (488 proteins) was found to be uniquely expressed between lipid accumulating phase and growth phase. Interestingly, there were up-regulated proteins involved in glycolysis, glycerolipid, carotenoid and glutathione metabolism which were preferable metabolic routes towards lipid accumulation and DHA production as well as cellular oxidative defence. Integrated proteomic and transcriptomic data were also conducted to comprehend the gene and protein regulation underlying the lipid and DHA biosynthesis. A significant up-regulation of acetyl-CoA synthetase was observed which suggests alternative route of acetate metabolism for acetyl-CoA producer. This study presents the holistic routes underlying lipid accumulation and DHA production in Aurantiochytrium sp. SW1 and other relevant thraustochytrid.


Sujet(s)
Acide docosahexaénoïque , Straménopiles , Acide docosahexaénoïque/métabolisme , Acétyl coenzyme A/métabolisme , Protéomique , Straménopiles/génétique , Straménopiles/métabolisme , Analyse de profil d'expression de gènes
19.
Biotechnol J ; 18(12): e2300314, 2023 Dec.
Article de Anglais | MEDLINE | ID: mdl-37596914

RÉSUMÉ

Docosahexaenoic acid (DHA) as one of ω-3 polyunsaturated fatty acids (PUFAs), plays a key role in brain development, and is widely used in food additives and the pharmaceutical industry. Schizochytrium sp. is often considered as a satisfactory strain for DHA industrialization. The aim of this study was to assess the feasibility of phosphopantetheinyl transferase (PPTase) and ω-3 fatty acid desaturase (FAD) for regulating DHA content in Schizochytrium sp. PPTase is essential to activate the polyketide-like synthase (PKS) pathway, which can transfer apo-acyl-carrier protein (apo-ACP) into holo-ACP, and plays a key role in DHA synthesis. Moreover, DHA and docosapentaenoic acid (DPA) are synthesized by the PKS pathway simultaneously, so high DPA synthesis limits the increase of DHA content. In addition, the detailed mechanisms of PKS pathway have not been fully elucidated, so it is difficult to improve DHA content by modifying PKS. However, ω-3 FAD can convert DPA into DHA, and it is the most direct and effective way to increase DHA content and reduce DPA content. Based on this, PPTase was overexpressed to enhance the synthesis of DHA by the PKS pathway, overexpressed ω-3 FAD to convert the co-product of the PKS pathway into DHA, and co-overexpressed PPTase and ω-3 FAD. With these strategies, compared with wild type, the final lipid, and DHA titer were 92.5 and 51.5 g L-1 , which increased by 46.4% and 78.1%, respectively. This study established an efficient DHA production strain, and provided some feasible strategies for industrial DHA production in Schizochytrium sp.


Sujet(s)
Acide docosahexaénoïque , Straménopiles , Acide docosahexaénoïque/métabolisme , Straménopiles/génétique , Straménopiles/métabolisme , Fatty acid desaturases/génétique , Fatty acid desaturases/métabolisme , Polyketide synthases/métabolisme
20.
Int J Mol Sci ; 24(14)2023 Jul 11.
Article de Anglais | MEDLINE | ID: mdl-37511067

RÉSUMÉ

The exposure of skin cells to UV radiation leads to redox imbalances and inflammation. The present study investigates a lipid extract obtained from the microalga Nannochloropsis oceanica as a potential protector against UVB-induced disturbances in human keratinocytes. The findings of this study show that the Nannochloropsis oceanica extract significantly inhibits UVB-induced cell death while concurrently decreasing the activity of pro-oxidative enzymes (xanthine and NADPH oxidase) and reducing the levels of ROS. Furthermore, the extract augments the activity of antioxidant enzymes (superoxide dismutases and catalase), as well as glutathione/thioredoxin-dependent systems in UVB-irradiated cells. The expression of Nrf2 factor activators (p62, KAP1, p38) was significantly elevated, while no impact was observed on Nrf2 inhibitors (Keap1, Bach1). The antioxidant activity of the extract was accompanied by the silencing of overexpressed membrane transporters caused by UVB radiation. Furthermore, the Nannochloropsis oceanica extract exhibited anti-inflammatory effects in UVB-irradiated keratinocytes by decreasing the levels of TNFα, 8-iso prostaglandin F2, and 4-HNE-protein adducts. In conclusion, the lipid components of Nannochloropsis oceanica extract effectively prevent the pro-oxidative and pro-inflammatory effects of UVB radiation in keratinocytes, thereby stabilizing the natural metabolism of skin cells.


Sujet(s)
Microalgues , Straménopiles , Humains , Microalgues/métabolisme , Stress oxydatif , Protéine-1 de type kelch associée à ECH/métabolisme , Rayons ultraviolets/effets indésirables , Facteur-2 apparenté à NF-E2/métabolisme , Kératinocytes/métabolisme , Antioxydants/pharmacologie , Straménopiles/métabolisme , Lipides/pharmacologie
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