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Three new strains of dissimilatory perchlorate-reducing bacteria (DPRB), QD19-16, QD1-5, and P3-1, were isolated from an active sludge. Phylogenetic trees based on 16S rRNA genes indicated that QD19-16, QD1-5, and P3-1 belonged to Brucella, Acidovorax, and Citrobacter, respectively, expanding the distribution of DPRB in the Proteobacteria. The three strains were gram-negative and facultative anaerobes with rod-shaped cells without flagella, which were 1.0-1.6 µm long and 0.5-0.6 µm wide. The three DPRB strains utilized similar broad spectrum of electron donors and acceptors and demonstrated a similar capability to reduce perchlorate within 6 days. The enzyme activity of perchlorate reductase in QD19-16 toward chlorate was higher than that toward perchlorate. The high sequence similarity of the perchlorate reductase operon and chlorite dismutase genes in the perchlorate reduction genomic islands (PRI) of the three strains implied that they were monophyletic origin from a common ancestral PRI. Two transposase genes (tnp1 and tnp2) were found in the PRIs of strain QD19-16 and QD1-5, but were absent in the strain P3-1 PRI. The presence of fragments of IR sequences in the P3-1 PRI suggested that P3-1 PRI had previously contained these two tnp genes. Therefore, it is plausible to suggest that a common ancestral PRI transferred across the strains Brucella sp. QD19-16, Acidovorax sp. QD1-5, and Citrobacter sp. P3-1 through horizontal gene transfer, facilitated by transposases. These results provided a direct evidence of horizontal gene transfer of PRI that could jump across phylogenetically unrelated bacteria through transposase. KEY POINTS: ⢠Three new DPRB strains can effectively remove high concentration of perchlorate. ⢠The PRIs of three DPRB strains are acquired from a single ancestral PRI. ⢠PRIs are incorporated into different bacteria genome through HGT by transposase.
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Ilhas Genômicas , Percloratos , Filogenia , Oxirredução , Transferência Genética Horizontal , RNA Ribossômico 16S/genética , Bactérias/genética , Oxirredutases/genética , Ecossistema , Transposases/genéticaRESUMO
Plastoglobules, which are lipoprotein structures surrounded by a single hydrophobic phospholipid membrane, are subcellular organelles in plant chromoplasts and chloroplasts. They contain neutral lipids, tocopherols, quinones, chlorophyll metabolites, carotenoids and their derivatives. Proteomic studies indicated that plastoglobules are involved in carotenoid metabolism and storage. In this study, one of the plastid lipid-associated proteins (PAP), the major protein in plastoglobules, was selected and overexpressed in Phaeodactylum tricornutum. The diameter of the plastoglobules in mutants was decreased by a mean of 19.2% versus the wild-type, while the fucoxanthin level was increased by a mean of 51.2%. All mutants exhibited morphological differences from the wild-type, including a prominent increase in the transverse diameter. Moreover, the unsaturated fatty acid levels were increased in different mutants, including an 18.9-59.3% increase in eicosapentaenoic acid content. Transcriptomic analysis revealed that PAP expression and the morphological changes altered xanthophyll synthesis and storage, which affected the assembly of the fucoxanthin chlorophyll a/c-binding protein and expression of antenna proteins as well as reduced the non-photochemical quenching activity of diatom cells. Therefore, metabolic regulation at the suborganelle level can be achieved by modulating PAP expression. These findings provide a subcellular structural site and target for synthetic biology to modify pigment and lipid metabolism in microalgae chassis cells.
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Nannochloropsis oceanica is widely used as a model photosynthetic chassis to produce fatty acids and carotenoid pigments. However, intense light typically causes excessive generation of reactive oxygen species (ROS) and photorespiration in microalgal cells, which results in decreased cell growth rate and unsaturated fatty acid content. In this study, the Vitreoscilla hemoglobin gene (vgb) was introduced into N. oceanica cells and expressed by using the light-harvesting complex promoter and its signal peptide. Compared with wild type (WT), the growth rate of transformants increased by 7.4%-18.5%, and the eicosapentaenoic acid content in an optimal transformant increased by 21.0%. Correspondingly, the intracellular ROS levels decreased by 56.9%-70.0%, and the catalase content in transformants was about 1.8 times that of WT. The photorespiration level of transformants was reduced by the measurement and calculation of the dissolved oxygen concentration under the condition of light-dark transition. The expression level of the key genes related to the photorespiration pathway in transformants was more than 80% lower than that in WT. These results indicated that Vitreoscilla hemoglobin could improve microalgal growth by reducing ROS damage and modulating photorespiration under stress conditions.
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Proteínas de Bactérias/metabolismo , Luz , Estramenópilas/metabolismo , Hemoglobinas Truncadas/metabolismo , Oxirredutases do Álcool/genética , Oxirredutases do Álcool/metabolismo , Proteínas de Bactérias/genética , Catalase/metabolismo , Complexos de Proteínas Captadores de Luz/genética , Fotossíntese/efeitos da radiação , Plasmídeos/genética , Plasmídeos/metabolismo , Regiões Promotoras Genéticas , Sinais Direcionadores de Proteínas/genética , Espécies Reativas de Oxigênio/metabolismo , Estramenópilas/efeitos da radiação , Hemoglobinas Truncadas/genéticaRESUMO
BACKGROUND: Chronic hypoxia plays an important role in the initiation and progression of chronic renal disease. The pathogenic role of chronic hypoxia in tubulointerstitial injury has been investigated widely, but little is known about acute hypoxia implications in glomerular damage. In this study, we investigated the effect of chronic hypoxia on transient receptor potential cation channel 6 (TRPC6) and the underlying mechanism in cultured human podocytes. METHODS: Fluo-3 was used as a calcium indicator of the OAG-induced receptor operated calcium entry (ROCE) and basal [Ca2+]i levels were monitored using laser scanning confocal microscope after exposure of cells to chronic hypoxia. 2-aminoethoxydiphenylborane (2-APB), a pharmacological blocker of TRPCs channels, was used to determine the role of TRPC6 in podocytes under chronic hypoxia. The mRNA expression and protein levels of TRPC6 were determined using Real-time RT-PCR and Western Blotting under normoxic and chronic hypoxic conditions. Actin arrangement was analyzed by confocal microscopy using phalloidin staining of F-actin in podocytes. RESULTS: Cytosolic free Ca2+ was increased by hypoxia or the treatment of TRPC6 agonist OAG under normoxic conditions. The increase of intracellular Ca2+ induced by hypoxia was time- and dose-dependent, which can be inhibited by 2-APB, demonstrating that the changes of intracellular Ca2+ induced by OAG depend on the activation of TRPC6. Further study showed that the TRPC6 expression levels were significantly increased by hypoxia, which were inhibited by the HIF1α inhibitor in podocytes. Similarly, the increase of intracellular Ca2+ induced by hypoxia was decreased when the podocytes were incubated with HIF1α inhibitor. We also found that F-actin was ruptured by hypoxia in podocytes, showing cytoskeleton reorganization. CONCLUSIONS: TRPC6 mRNA and protein expression levels were significantly increased in podocytes under hypoxia, which may result in the increase of intracellular Ca2+. This alternation of TRPC6 may be relevant to the modulation of HIF1α. Hypoxia in podocytes can result in cytoskeleton reorganization, which further leads to podocytes injury and disfunction.
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Subunidade alfa do Fator 1 Induzível por Hipóxia/metabolismo , Hipóxia/genética , Hipóxia/metabolismo , Podócitos/metabolismo , Podócitos/patologia , Canal de Cátion TRPC6/genética , Canal de Cátion TRPC6/metabolismo , Actinas/metabolismo , Compostos de Boro/farmacologia , Cálcio/metabolismo , Sinalização do Cálcio/efeitos dos fármacos , Linhagem Celular , Citoesqueleto/metabolismo , Diglicerídeos/farmacologia , Humanos , Subunidade alfa do Fator 1 Induzível por Hipóxia/antagonistas & inibidores , Oxigênio/metabolismo , Oxigênio/farmacologia , Podócitos/efeitos dos fármacos , RNA Mensageiro/análise , RNA Mensageiro/genética , Fatores de TempoRESUMO
Background: Adolescents differ from adults in certain aspects, but the differences in the etiology of chronic cough between adolescents and adults have not been specifically investigated. The purpose of this study is to evaluate the causes of chronic cough in adolescent patients in comparison with those in adult patients. Methods: The electronic medical records were retrospectively screened for patients with the initial diagnosis of chronic cough from 2016 to 2018. Clinical variables and the causes of chronic cough were collected in patients who met inclusion criteria. Patients were assigned to adolescent (13 to 18 years) and adult (> 18 years) group based on age. The distribution of causes of chronic cough was compared by t-test and χ2 tests between adolescent and adult groups using SPSS. Significant level was set at 0.05. Results: The enrolled patients consisted of 16 adolescents and 73 adults with chronic cough. The most common causes of chronic cough were cough-variant asthma and upper airway cough syndrome in both adolescents and adults. The frequency of post infectious cough in adolescents was significantly higher than that in adults (12.5% vs. 1.36%; p= 0.024). Although no significant difference between the two groups was found, there was a trend that suggested that habitual cough was more common in the adolescent group (6.25%:1.36%); while gastroesophageal reflux disease (GERD) did not show similar results (6.25%:20.54%). Conclusion: There are moderate differences in the causes of chronic cough between adolescent and adult patients. Realizing those differences would be helpful for clinicians to establish an appropriate differential diagnosis and make referral decisions.
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The aims of the present study were to investigate the roles and underlying mechanisms of microRNA1243p (miR1243p) in the progression of acute myocardial infarction (AMI). The expression of miR1243p was determined via reverse transcriptionquantitative polymerase chain reaction (RTqPCR). TargetScan analysis and a luciferase reporter assay were conducted to reveal the association between miR1243p and nuclear factor κlightchainenhancer of activated B cells (NFκB) repressing factor (NKRF). To investigate the role of miR1243p in AMI, a cell model of myocardial hypoxic/ischemic injury was established by subjecting H9c2 cardiac cells to hypoxia for 48 h. The viability of cells was determined using an MTT assay, and cell apoptosis was analyzed by flow cytometry. Additionally, the expression levels of inflammatory factors [tumor necrosis factorα (TNFα), interleukin (IL)1ß and IL6] were measured via ELISA. Furthermore, gene and protein expression levels were determined by performing RTqPCR and western blot analyses, respectively. It was revealed that the expression of miR1243p was significantly increased in the blood of patients with AMI and hypoxiatreated H9c2 cells. Additionally, it was demonstrated that NKRF was a direct target of miR1243p. The hypoxiainduced decrease in the viability of H9c2 cells and increase in cell apoptosis were eliminated by the downregulation of miR1243p. Furthermore, hypoxia significantly increased the levels of TNFα, IL1ß and IL6, whereas miR1243p downregulation eliminated these effects. Downregulated expression of Bcell lymphoma 2, procaspase 3 and procaspase 9 protein, and upregulated expression of cleaved caspases 3 and 9 was observed in hypoxic H9c2 cells; the altered expression of these proteins was suppressed by miR1243p inhibitor. Additionally, miR1243p inhibitor suppressed the hypoxiainduced activation of the NFκB signaling pathway in H9c2 cells. Furthermore, it was demonstrated that the various effects of miR1243p inhibitor on H9c2 cells were eliminated by the small interfering RNAmediated downregulation of NKRF. In conclusion, the results of the present study indicated that miR1243p downregulation protected against AMI via inhibition of inflammatory responses and the apoptosis of cardiomyocytes by regulating the NKRF/NFκB pathway.
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Apoptose , MicroRNAs/metabolismo , Infarto do Miocárdio/metabolismo , Miócitos Cardíacos/metabolismo , Adulto , Animais , Proteínas Reguladoras de Apoptose/metabolismo , Linhagem Celular , Citocinas/metabolismo , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Infarto do Miocárdio/patologia , Infarto do Miocárdio/prevenção & controle , Miócitos Cardíacos/patologia , RatosRESUMO
Cellular senescence is a hallmark of aging, it is a permanent state of cell cycle arrest induced by cellular stresses. During the aging process, senescent cells (SCs) increasingly accumulate in tissues, causing a loss of tissue-repair capacity because of cell cycle arrest in progenitor cells and produce proinflammatory and matrix-degrading molecules which are known as the senescence-associated secretory phenotype (SASP), and thereby contribute to the development of various age-related diseases. Genetic evidence has demonstrated that clearance of SCs can delay aging and extend healthspan. Senolytics, small molecules that can selectively kill SCs, have been developed to treat various age-related diseases. In recent years, emerging natural compounds have been discovered to be effective senolytic agents, such as quercetin, fisetin, piperlongumine and the curcumin analog. Some of the compounds have been validated in animal models and have great potential to be pushed to clinical applications. In this review, we will discuss cellular senescence and its potential as a target for treating age-related diseases, and summarize the known natural compounds as senolytic agents and their applications.
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Envelhecimento/efeitos dos fármacos , Pontos de Checagem do Ciclo Celular/efeitos dos fármacos , Senescência Celular/efeitos dos fármacos , Citotoxinas/uso terapêutico , Envelhecimento/fisiologia , Animais , Pontos de Checagem do Ciclo Celular/fisiologia , Senescência Celular/fisiologia , Curcumina/análogos & derivados , Curcumina/química , Curcumina/uso terapêutico , Citotoxinas/química , Dioxolanos/química , Dioxolanos/uso terapêutico , Flavonoides/química , Flavonoides/uso terapêutico , Flavonóis , Humanos , Quercetina/química , Quercetina/uso terapêuticoRESUMO
Mychonastes afer HSO-3-1 is a potential producer of nervonic acid, which could be accumulated to 2-3% of dry cell weight. Improving the productivity of nervonic acid is critical to promote the commercialization of this product. In this study, 1-naphthylacetic acid (NAA) and tea polyphenol (TP) were selected as bioactive additives to stimulate the growth of M. afer. Supplementing NAA in the early growth stage and TP in the middle and late growth stage led to improved lipid accumulation in M. afer. The cultures supplemented with TP at the late growth stage maintained higher photosynthetic efficiency than the control groups without TP. Furthermore, the intracellular reactive oxygen species (ROS) accumulations in M. afer supplemented with 500 mg/L of TP was 63% lower than the control group. A linear relationship (R2= 0.899) between the values of Fv/Fm and ROS accumulation was established. We hypothesize supplement of bioactive additives at different growth stage could promote the cell growth rate and nervonic acid productivity of M. afer by retrieving intracellular ROS level. Further analysis of photosynthetic system II (PSII) protein in M. afer cultured in presence of NAA and TP indicated the levels of D1 and D2 proteins, the core skeleton proteins of PSII, showed 33.3 and 25.6% higher than the control group. CP43 protein, a critical module in PSII repair cycle, decreased significantly. These implied that TP possesses the function of slowing down the damage of PSII by scavenging excess intracellular ROS.
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Nervonic acid (NA) is a major very long-chain monounsaturated fatty acid found in the white matter of mammalian brains, which plays a critical role in the treatment of psychotic disorders and neurological development. In the nature, NA has been synthesized by a handful plants, fungi, and microalgae. Although the metabolism of fatty acid has been studied for decades, the biosynthesis of NA has yet to be illustrated. Generally, the biosynthesis of NA is considered starting from oleic acid through fatty acid elongation, in which malonyl-CoA and long-chain acyl-CoA are firstly condensed by a rate-limiting enzyme 3-ketoacyl-CoA synthase (KCS). Heterologous expression of kcs gene from high NA producing species in plants and yeast has led to synthesis of NA. Nevertheless, it has also been reported that desaturases in a few plants can catalyze very long-chain saturated fatty acid into NA. This review highlights recent advances in the biosynthesis, the sources, and the biotechnological aspects of NA.
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Biotecnologia/métodos , Ácidos Graxos Monoinsaturados/metabolismo , Microalgas/metabolismo , Acil Coenzima A/metabolismo , Animais , Biotecnologia/tendências , Microbiologia Industrial/tendências , Malonil Coenzima A/metabolismo , Microalgas/genéticaRESUMO
Fucoxanthin is a natural pigment found in microalgae, especially diatoms and Chrysophyta. Recently, it has been shown to have anti-inflammatory, anti-tumor, and anti-obesityactivity in humans. Phaeodactylum tricornutum is a diatom with high economic potential due to its high content of fucoxanthin and eicosapentaenoic acid. In order to improve fucoxanthin production, physical and chemical mutagenesis could be applied to generate mutants. An accurate and rapid method to assess the fucoxanthin content is a prerequisite for a high-throughput screen of mutants. In this work, the content of fucoxanthin in P. tricornutum was determined using spectrophotometry instead of high performance liquid chromatography (HPLC). This spectrophotometric method is easier and faster than liquid chromatography and the standard error was less than 5% when compared to the HPLC results. Also, this method can be applied to other diatoms, with standard errors of 3-14.6%. It provides a high throughput screening method for microalgae strains producing fucoxanthin.
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Diatomáceas/química , Espectrofotometria/métodos , Xantofilas/química , Cromatografia Líquida/métodos , Diatomáceas/genética , Mutagênese/genéticaRESUMO
Curcumin is an attractive agent due to its multiple bioactivities. However, the low oral bioavailability and efficacy profile hinders its clinical application. To improve the bioavailability, many analogs of curcumin have been developed, among which EF24 is an excellent representative. EF24 has enhanced bioavailability over curcumin and shows more potent bioactivity, including anti-cancer, anti-inflammatory, and anti-bacterial. EF24 inhibits tumor growth by inducing cell cycle arrest and apoptosis, mainly through its inhibitory effect on the nuclear factor kappa B (NF-κB) pathway and by regulating key genes through microRNA (miRNA) or the proteosomal pathway. Based on the current structure, more potent EF24 analogs have been designed and synthesized. However, some roles of EF24 remain unclear, such as whether it induces or inhibits reactive oxygen species (ROS) production and whether it stimulates or inhibits the mitogen activated kinase-like protein (MAPK) pathway. This review summarizes the known biological and pharmacological activities and mechanisms of action of EF24.
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Edible oil industry produced massive wastewater, which requires extensive treatment to remove pungent smell, high phosphate, carbon oxygen demand (COD), and metal ions prior to discharge. Traditional anaerobic and aerobic digestion could mainly reduce COD of the wastewater from oil refinery factories (WEORF). In this study, a robust oleaginous microalga Desmodesmus sp. S1 was adapted to grow in WEORF. The biomass and lipid content of Desmodesmus sp. S1 cultivated in the WEORF supplemented with sodium nitrate were 5.62 g·L(-1) and 14.49%, whereas those in the WEORF without adding nitrate were 2.98 g·L(-1) and 21.95%. More than 82% of the COD and 53% of total phosphorous were removed by Desmodesmus sp. S1. In addition, metal ions, including ferric, aluminum, manganese and zinc were also diminished significantly in the WEORF after microalgal growth, and pungent smell vanished as well. In comparison with the cells grown in BG-11 medium, the cilia-like bulges and wrinkles on the cell surface of Desmodesmus sp. S1 grown in WEORF became out of order, and more polyunsaturated fatty acids were detected due to stress derived from the wastewater. The study suggests that growing microalgae in WEORF can be applied for the dual roles of nutrient removal and biofuel feedstock production.
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Clorófitas/metabolismo , Microalgas/metabolismo , Óleos de Plantas/análise , Águas Residuárias/análise , Poluentes Químicos da Água/metabolismo , Biodegradação Ambiental , Análise da Demanda Biológica de OxigênioRESUMO
A large proportion of mutants with altered pigment features have been obtained via exposure to heavy-ion beams, a technique that is efficient for trait improvement in the breeding of plants and algae. However, little is known about the underlying mechanisms by which the photosynthetic pigments are altered by heavy-ion irradiation. In our study, the photosynthetic characteristics of progenies from carbon-ion irradiated Selenastrum capricornutum were investigated. Five progenies deficient in chlorophyll a were isolated after carbon-ion exposure. Photosynthetic characteristics, photoprotection capacity and gene expression of the light-harvesting complex in these progenies were further characterized by the measurement of chlorophyll fluorescence parameters (Fv/Fm, ФPSII, NPQ, ETR), the de-epoxidation state of the xanthophyll cycle, the amount of lutein and quantitative real-time PCR. High maximum quantum yield of photosystem II at day 10 and high thermal dissipation ability were observed in progenies #23 and #37 under normal culture condition. Progenies #18, #19 and #20 showed stronger resistance against high levels of light steps than the control group (612-1077 µmol photons m -2 s -1, p< 0.05). The progenies #20 and #23 exhibited strong photoprotection by thermal dissipation and quenching of 3Chl* after 24 h of high light treatment. The mRNA levels of Lhcb5, Lhcbm5 and Lhcbm1 of the light-harvesting complex revealed markedly differential expression in the five progenies irradiated by carbon-ion beams. This work indicates that photosynthetic efficiency, photoprotection ability and the expression of light-harvesting antennae in unicellular green algae can be markedly influenced by irradiation. To our knowledge, this is the first report on changes in the photosynthetic pigments of green algae after treatment with carbon-ion beams.
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Isótopos de Carbono , Clorófitas/metabolismo , Fotossíntese/efeitos da radiação , Clorofila/genética , Clorofila/metabolismo , Clorofila A , Clorófitas/genética , Clorófitas/efeitos da radiação , Expressão Gênica/efeitos da radiação , Complexos de Proteínas Captadores de Luz/metabolismo , Mutação , Fotossíntese/genética , Complexo de Proteína do Fotossistema II/metabolismo , Xantofilas/metabolismoRESUMO
In order to relieve the pressure of energy supply and environment contamination that humans are facing, there are now intensive worldwide efforts to explore natural bioresources for production of energy storage compounds, such as lipids, alcohols, hydrocarbons, and polysaccharides. Around the world, many plants have been evaluated and developed as feedstock for bioenergy production, among which several crops have successfully achieved industrialization. Microalgae are another group of photosynthetic autotroph of interest due to their superior growth rates, relatively high photosynthetic conversion efficiencies, and vast metabolic capabilities. Heterotrophic microorganisms, such as yeast and bacteria, can utilize carbohydrates from lignocellulosic biomass directly or after pretreatment and enzymatic hydrolysis to produce liquid biofuels such as ethanol and butanol. Although finding a suitable organism for biofuel production is not easy, many naturally occurring organisms with good traits have recently been obtained. This review mainly focuses on the new organism resources discovered in the last 5 years for production of transport fuels (biodiesel, gasoline, jet fuel, and alkanes) and hydrogen, and available methods to improve natural organisms as platforms for the production of biofuels.
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Fenômenos Fisiológicos Bacterianos , Biocombustíveis/microbiologia , Conservação dos Recursos Naturais/métodos , Fungos/fisiologia , Microalgas/fisiologia , Fenômenos Fisiológicos Vegetais , Biodiversidade , Microalgas/classificaçãoRESUMO
Since CO2 concentrations in industrial flue gases are usually 10%-20%, one of the prerequisites for efficient CO2 removal by algae is the level of tolerance of microalgal species to exposure to high concentrations of CO2. A newly isolated microalgal strain, Chlorella sp. MRA-1, could retain growth with high concentrations of CO2 up to 15%. The highest lipid productivity for Chlorella sp. MRA-1 was 0.118 g/l/day with a 5% CO2 concentration. Octadecenoic acid and hexadecanoic acid, the main components of biodiesel, accounted for 70% of the total fatty acids. A lipid content of 52% of dry cell weight was achieved with limited amounts of nitrogen. Chlorella sp. MRA-1 seems to be an ideal candidate for biodiesel production when cultured with high concentrations of CO2.
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Adaptação Biológica , Dióxido de Carbono/metabolismo , Chlorella/metabolismo , Lipídeos/biossíntese , Chlorella/química , Chlorella/crescimento & desenvolvimento , Meios de Cultura/química , Ácidos Graxos/biossíntese , Ácidos Graxos/química , Nitrogênio/metabolismoRESUMO
The unicellular green microalga Desmodesmus sp. S1 can produce more than 50% total lipid of cell dry weight under high light and nitrogen-limitation conditions. After irradiation by heavy (12)C(6+) ion beam of 10, 30, 60, 90 or 120 Gy, followed by screening of resulting mutants on 24-well microplates, more than 500 mutants were obtained. One of those, named D90G-19, exhibited lipid productivity of 0.298 g L(-1)â d(-1), 20.6% higher than wild type, likely owing to an improved maximum quantum efficiency (Fv/Fm) of photosynthesis under stress. This work demonstrated that heavy-ion irradiation combined with high-throughput screening is an effective means for trait improvement. The resulting mutant D90G-19 may be used for enhanced lipid production.
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Carbono/metabolismo , Íons Pesados , Lipídeos/biossíntese , Microalgas/metabolismo , Fotossíntese , Biomassa , Sobrevivência Celular/genética , Sobrevivência Celular/efeitos da radiação , Ácidos Graxos/química , Ácidos Graxos/metabolismo , Genes Letais , Lipídeos/química , Microalgas/genética , Microalgas/crescimento & desenvolvimento , Mutação , Taxa de Mutação , Fenótipo , Amido/metabolismoRESUMO
Scenedesmus spp. have been reported as potential microalgal species used for the lipid production. This study investigated the effects of light intensity (at three levels: 50, 250, and 400 µmol photons m(-2) s(-1)) on the growth and lipid production of Scenedesmus sp. 11-1 under N-limited condition. Carotenoid to chlorophyll ratio was higher when algae 11-1 grew under 250 and 400 µmol photons m(-2) s(-1) than that under 50 µmol photons m(-2) s(-1), while protein contents was lower. Highest biomass yield (3.88 g L(-1)), lipid content (41.1 %), and neutral lipid content (32.9 %) were achieved when algae 11-1 grew at 400 µmol photons m(-2) s(-1). Lipid production was slight lower at 250 µmol photons m(-2) s(-1) level compared to 400 µmol photons m(-2) s(-1). The major fatty acids in the neutral lipid of 11-1 were oleic acid (43-52 %), palmitic acid (24-27 %), and linoleic acid (7-11 %). In addition, polyunsaturated fatty acids had a positive correlation with total lipid production, and monounsaturated fatty acids had a negative one.
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Luz , Metabolismo dos Lipídeos/efeitos da radiação , Microalgas/crescimento & desenvolvimento , Microalgas/metabolismo , Nitrogênio/deficiência , Scenedesmus/crescimento & desenvolvimento , Scenedesmus/metabolismo , Relação Dose-Resposta à Radiação , Ácidos Graxos/química , Ácidos Graxos/metabolismo , Microalgas/efeitos da radiação , Nitratos/metabolismo , Pigmentos Biológicos/metabolismo , Proteínas de Plantas/metabolismo , Scenedesmus/efeitos da radiaçãoRESUMO
BACKGROUND: Biodiesel is considered to be a promising future substitute for fossil fuels, and microalgae are one source of biodiesel. The ratios of lipid, carbohydrates and proteins are different in different microalgal species, and finding a good strain for oil production remains a difficult prospect. Strains producing valuable co-products would improve the viability of biofuel production. RESULTS: In this study, we performed sequence analysis of the 18S rRNA gene and internal transcribed spacer (ITS) of an algal strain designated HSO-3-1, and found that it was closely related to the Mychonastes afer strain CCAP 260/6. Morphology and cellular structure observation also supported the identification of strain HSO-3-1 as M. afer. We also investigated the effects of nitrogen on the growth and lipid accumulation of the naturally occurring M. afer HSO-3-1, and its potential for biodiesel production. In total, 17 fatty acid methyl esters (FAMEs) were identified in M. afer HSO-3-1, using gas chromatography/mass spectrometry. The total lipid content of M. afer HSO-3-1 was 53.9% of the dry cell weight, and we also detected nervonic acid (C24:1), which has biomedical applications, making up 3.8% of total fatty acids. The highest biomass and lipid yields achieved were 3.29 g/l and 1.62 g/l, respectively, under optimized conditions. CONCLUSION: The presence of octadecenoic and hexadecanoic acids as major components, with the presence of a high-value component, nervonic acid, renders M. afer HSO-3-1 biomass an economic feedstock for biodiesel production.
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Starch and neutral lipids are two major carbon storage compounds in many microalgae and plants. Lipids are more energy rich and have often been used as food and fuel feedstocks. Genetic engineering of the lipid biosynthesis pathway to overproduce lipid has achieved only limited success. We hypothesize that through blocking the competing pathway to produce starch, overproduction of neutral lipid may be achieved. This hypothesis was tested using the green microalga Chlamydomonas reinhardtii and its low starch and starchless mutants. We discovered that a dramatic increase in neutral lipid content and the neutral lipid/total lipid ratio occurred among the mutants under high light and nitrogen starvation. BAFJ5, one of the mutants defective in the small subunit of ADP-glucose pyrophosphorylase, accumulated neutral and total lipid of up to 32.6% and 46.4% of dry weight (DW) or 8- and 3.5-fold higher, respectively, than the wild-type. These results confirmed the feasibility of increasing lipid production through redirecting photosynthetically assimilated carbon away from starch synthesis to neutral lipid synthesis. However, some growth impairment was observed in the low starch and starchless mutants, possibly due to altered energy partitioning in PSII, with more excitation energy dissipated as heat and less to photochemical conversion. This study demonstrated that biomass and lipid production by the selected mutants can be improved by physiological manipulation.
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Chlamydomonas reinhardtii/metabolismo , Metabolismo dos Lipídeos , Amido/metabolismo , Proteínas de Algas/genética , Biomassa , Chlamydomonas reinhardtii/enzimologia , Chlamydomonas reinhardtii/genética , Glucose-1-Fosfato Adenililtransferase/deficiência , Luz , Redes e Vias Metabólicas/genética , Mutação , Nitrogênio/metabolismoRESUMO
BACKGROUND AND OBJECTIVES: Invasive pulmonary aspergillosis (IPA) among patients with chronic obstructive pulmonary disease (COPD) is increasing in frequency. We conducted this study to find out the approximate incidence of IPA in patients with acute exacerbation of COPD (AECOPD), and to determine whether using a combination of two tests (galactomannan [GM] antigen and 1,3-beta-glucan [BG] detection) would result in a more specific diagnosis of IPA. METHODS: The study included 261 patients with AECOPD admitted over two years. The patients were categorized according to the modified definitions for IPA. GM antigen and BG were detected by the Platelia Aspergillus and Glucatell tests. RESULTS: Two patients had proven IPA, three had probable IPA, and two had possible IPA. The rate of proven or probable IPA in patients with AECOPD was 1.91% (5/261). Four patients with proven and probable IPA had been treated with a systemic or inhaled corticosteroid before hospitalization and the typical symptoms and diagnostic signs of IPA were relatively less common in them. Mortality in patients with proven and probable IPA was 80%. The specificity of combined GM and BG detection was 98.8%. CONCLUSION: Combining two serological tests increased the specificity of diagnosis but further trials are needed to prove the value of this approach.