58 research outputs found

    Phenotypic screening identifies Brefeldin A/Ascotoxin as an inducer of lipid storage in the algae \u3ci\u3eChlamydomonas reinhardtii\u3c/i\u3e

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    The use of microalgae as a biofuel feedstock is highly desired, but current methods to induce lipid accumulation cause severe stress responses that limit biomass and, thus oil yield. To address these issues, a high throughput screening (HTS) method was devised to identify chemical inducers of growth and lipid accumulation. Optimization was performed to determine the most effective cell density, DMSO and Nile Red (NR) concentrations to monitor growth and lipid accumulation. The method was tested using 1717 compounds from National Cancer Institute (NCI) Diversity Set III and Natural Products Set II in Chlamydomonas reinhardtii. Cells were inoculated at low density and 10 μM of the test compound was added. After 72 h, cell density was measured at OD550 and lipid accumulation assessed using NR fluorescence. Primary screening identified 8 compounds with a hit rate of 0.47% and a robust Z′ discrimination factor (0.68 ± 0.1). Of these, Brefeldin A (BFA) was the most successful at inducing lipid accumulation and was used to evaluate secondary screens including measuring levels of fatty acids, photosynthetic pigments, proteins and carbohydrates. The effectiveness of BFA was confirmed in Chlorella sorokiniana UTEX 1230. This study demonstrates the power of chemical genomics approaches in biofuel research

    Remodeling of Chlamydomonas Metabolism Using Synthetic Inducers Results in Lipid Storage during Growth

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    Microalgae accumulate lipids during stress such as that of nutrient deprivation, concomitant with cessation of growth and depletion of chloroplasts. By contrast, certain small chemical compounds selected by high-throughput screening in Chlamydomonas reinhardtii can induce lipid accumulation during growth, maintaining biomass. Comprehensive pathway analyses using proteomics, transcriptomics, and metabolomics data were acquired from Chlamydomonas cells grown in the presence of one of two structurally distinct lipid activators. WD10784 stimulates both starch and lipid accumulation, whereas WD30030-treated cells accumulate only lipids. The differences in starch accumulation are largely due to differential effects of the two compounds on substrate levels that feed into starch synthesis and on genes encoding starch metabolic enzymes. The compounds had differential effects on photosynthesis, respiration, and oxidative stress pathways. Cells treated with WD10784 showed slowed growth over time and reduced abundance of photosynthetic proteins, decreased respiration, and increased oxidative stress proteins, glutathione, and reactive oxygen species specific to this compound. Both compounds maintained central carbon and nitrogen metabolism, including the tricarboxylic acid cycle, glycolysis, respiration, and the Calvin–Benson–Bassham cycle. There were few changes in proteins and transcripts related to fatty acid biosynthesis, whereas proteins and transcripts for triglyceride production were elevated, suggesting that lipid synthesis is largely driven by substrate availability. This study reports that the compound WD30030 and, to a lesser extent WD10784, increases lipid and lipid droplet synthesis and storage without restricting growth or biomass accumulation by mechanisms that are substantially different from nutrient deprivation

    Parameters Affecting the Biosynthesis of Gold Nanoparticles Using the Aquatic Extract of Scrophularia striata and their Antibacterial Properties

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    Green synthesis is a simple, low-cost, non-toxic, environmentally friendly and efficient approach to use. Leaf extract of plants rich in polyphenols, such as flavonoids, is a powerful agent in reducing the synthesis of gold nanoparticles. The purpose of this study is to investigate the parameters affecting the biosynthesis of gold nanoparticles using the aqueous extract of Scrophularia striata plant and their antimicrobial activity. Biosynthesis of gold nanoparticles was accomplished by the interaction of golden salt (HAuCl4, 3H(2)O) with aqueous extract of Scrophularia striata. In order to obtain uniform and spherical nanoparticles, the following parameters affecting the biosynthesis of nanoparticles were investigated and optimized by ultraviolet-spectrophotometric technique; golden salt concentration, extract volume, pH and reaction time. Transmission electron microscopy and X-ray diffraction technique were also used to further characterize nanoparticles. Finally, the anti-bacterial properties of gold nanoparticles were investigated by disc diffusion method. The resulting absorption spectra exhibited strong peaks at 570 nm, which is a specific wavelength for gold nanoparticles. Transmission electron microscopy studies showed that the gold nanoparticles had a spherical shape with a mean diameter of 5-10nm, and the highest diameter of the growth inhibition zone was observed on the diameter of the hafnium bacteria (14mm). In this study, it was observed that, with the aid of Scrophularia striata aqueous extracts, a golden nanoparticle showed an antibacterial activity against gram-negative bacteria

    Organochlorine pesticides and polychlorinated biphenyls in city drains in Makurdi, central Nigeria : seasonal variations, source apportionment and risk assessment

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    A study of seasonal variation, sources and potential risks of organochlorine pesticides (OCPs) and polychlorinated biphenyls (PCBs) in open city drains in Makurdi, Central Nigeria was carried out. OCPs and PCBs were quantified using gas chromatograph-mass spectrometer. The total (∑8OCPs) concentrations (ngL−1) of OCPs in water was 2.99 with a mean ± SD of 0.75 ± 0.12 during wet season, while during dry season, the values were 11.43 and 2.86 ± 1.54 respectively. In sediment, the total concentration (ngg−1) of OCPs was 5270.66 with a mean ± SD of 1756.89 ± 450.01 during wet season and a total concentration of 5837.93 and the mean ± SD of 1945.98 ± 646.04, during dry season. Source apportionment of OCPs suggested historic application of the pollutants. The total (∑7PCBs) concentration (ngL−1) of PCBs in water was 0.24 with a mean ± SD of 0.03 ± 0.02 during wet season and a total concentration of 0.61 with a mean ± SD of 0.09 ± 0.11 during dry season. The total concentration (ngg−1) of PCBs in sediment was 37.88, mean ± SD of 5.41 ± 5.93 during wet season and a total of 47.07 and mean ± SD of 6.72 ± 7.27 during dry season. Ecological risk assessment based on effect range low (ERL) and effect range median (ERM) or threshold effect level (TEL) and probable effect level (PEL) that ecological risks were possible for some OCPs in this study, which calls for source control and remediation of the affected sites. Toxicity equivalency (TEQ) of PCB-118, the dioxin-like congener, indicated that it was most harmful to humans/mammals followed by birds, then fish

    Organochlorine pesticides and polychlorinated biphenyls in city drains in Makurdi, central Nigeria: Seasonal variations, source apportionment and risk assessment

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    A study of seasonal variation, sources and potential risks of organochlorine pesticides (OCPs) and polychlorinated biphenyls (PCBs) in open city drains in Makurdi, Central Nigeria was carried out. OCPs and PCBs were quantified using gas chromatograph-mass spectrometer. The total (∑8OCPs) concentrations (ngL−1) of OCPs in water was 2.99 with a mean ± SD of 0.75 ± 0.12 during wet season, while during dry season, the values were 11.43 and 2.86 ± 1.54 respectively. In sediment, the total concentration (ngg−1) of OCPs was 5270.66 with a mean ± SD of 1756.89 ± 450.01 during wet season and a total concentration of 5837.93 and the mean ± SD of 1945.98 ± 646.04, during dry season. Source apportionment of OCPs suggested historic application of the pollutants. The total (∑7PCBs) concentration (ngL−1) of PCBs in water was 0.24 with a mean ± SD of 0.03 ± 0.02 during wet season and a total concentration of 0.61 with a mean ± SD of 0.09 ± 0.11 during dry season. The total concentration (ngg−1) of PCBs in sediment was 37.88, mean ± SD of 5.41 ± 5.93 during wet season and a total of 47.07 and mean ± SD of 6.72 ± 7.27 during dry season. Ecological risk assessment based on effect range low (ERL) and effect range median (ERM) or threshold effect level (TEL) and probable effect level (PEL) that ecological risks were possible for some OCPs in this study, which calls for source control and remediation of the affected sites. Toxicity equivalency (TEQ) of PCB-118, the dioxin-like congener, indicated that it was most harmful to humans/mammals followed by birds, then fish
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