Structure and properties of oil bodies in diatoms Maeda, Yoshiaki; Nojima, Daisuke; Yoshino, Tomoko ...
Philosophical transactions - Royal Society. Biological sciences,
09/2017, Letnik:
372, Številka:
1728
Journal Article
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Diatoms accumulate triacylglycerols in spherical organelles called oil bodies when exposed to nutrient deprivation conditions. Oil body biology in diatoms has attracted significant attention due to ...the complexity of the intracellular organelles and the unique combination of genes generated by the evolutionary history of secondary endosymbiosis. The demand for biofuel production has further increased the interest in and importance of a better understanding of oil body biology in diatoms, because it could provide targets for genetic engineering to further enhance their promising lipid accumulation. This review describes recent progress in studies of the structure and properties of diatom oil bodies. Firstly, the general features of diatom oil bodies are described, in particular, their number, size and morphology, as well as the quantity and quality of lipids they contain. Subsequently, the diatom oil body-associated proteins, which were recently discovered through oil body proteomics, are introduced. Then, the metabolic pathways responsible for the biogenesis and degradation of diatom oil bodies are summarized. During biogenesis and degradation, oil bodies interact with other organelles, including chloroplasts, the endoplasmic reticulum and mitochondria, suggesting their dynamic nature in response to environmental changes. Finally, the functions of oil bodies in diatoms are discussed.
This article is part of the themed issue ‘The peculiar carbon metabolism in diatoms’.
Wastewater treatment, along with the simultaneous production of valuable chemical compounds, including lipids by microalgae is a challenging but attractive study. Towards this goal, the candidate ...microalgae were selected from culture collections or isolated from wastewater in this study. The initial screening test using microalgae revealed that various eukaryotic as well as prokaryotic microalgae showed steady growth in municipal wastewater samples. Among them, Tetraselmis sp. NKG400013 and Parachlorella kessleri NKG021201 from culture collections, and Chloroidium saccharophilum NKH13 from the wastewater sample exhibited high biomass productivity. Furthermore, P. kessleri NKG021201 and C. saccharophilum NKH13 showed high lipid productivity (56 ± 1 mg/L/day for NKG021201, 35 ± 10 mg/L/day for NKH13). During this cultivation, 99% of nitrogen and 82% of phosphorous compounds were removed from the wastewater sample by the strain NKG021201. Analysis of fatty acid compositions of P. kessleri NKG021201 and C. saccharophilum NKH13 revealed that lipids derived from these microalgae were suitable for the application of biodiesel fuels, indicating that these microalgae were promising for wastewater treatment and lipid production.
Circulating tumor cells (CTCs) are tumor cells circulating in the peripheral blood of patients with metastatic cancer. Detection of CTCs has clinical significance in cancer therapy because it would ...enable earlier diagnosis of metastasis. In this research, a microfluidic device equipped with a size-selective microcavity array for highly efficient and rapid detection of tumor cells from whole blood was developed. The microcavity array can specifically separate tumor cells from whole blood on the basis of differences in the size and deformability between tumor and hematologic cells. Furthermore, the cells recovered on the microcavity array were continuously processed for image-based immunophenotypic analysis using a fluorescence microscope. Our device successfully detected approximately 97% of lung carcinoma NCI-H358 cells in 1 mL whole blood spiked with 10−100 NCI-H358 cells. In addition, breast, gastric, and colon tumor cells lines that include EpCAM-negative tumor cells, which cannot be isolated by conventional immunomagnetic separation, were successfully recovered on the microcavity array with high efficiency (more than 80%). On an average, approximately 98% of recovered cells were viable. Our microfluidic device has high potential as a tool for the rapid detection of CTCs and can be used to study CTCs in detail.
Omega-3 polyunsaturated fatty acids (PUFAs) such as eicosapentaenoic acid (EPA, C20:5ω-3) are essential for human health and fish growth especially in aquaculture sector. However, with the growing of ...aquaculture, the demand of PUFA supply also has been increasing. Fistulifera solaris, a marine diatom, is known for its ability to accumulate 65% of lipid content per dry cell weight, and can produce the high content of EPA. Thus, this diatom shows a great potential to be a feedstock of omega-3 PUFAs for fish feeds. In this study, in order to further understand and enhance the metabolism of PUFA biosynthesis in the diatom, the impacts of ketoacyl-ACP synthase (KAS) and ketoacyl-CoA synthase (KCS) inhibition on the PUFA production were analyzed by adding the specific inhibitors. KAS and KCS enzymes both play a role in the fatty acid elongation. As a result, the inhibition of KAS showed an increase in EPA content without arresting the cell growth. On the other hand, inhibition of KCS did not show a significant impact on the PUFA content in F. solaris. Our finding suggests that the specific suppression of KAS function can be a promising way to enhance the omega-3 PUFA production in F. solaris.
The immobilization of multiple cellulase complexes receiving attention for use in the efficient hydrolysis of celluloses. In this study, the magnetosome display system was employed for the ...preparation of systems mimicking natural multiple cellulase complexes (cellulosomes) on magnetic nanoparticles (MNPs). Initially, two fluorescent proteins, namely, green fluorescent protein and mCherry, were immobilized on MNPs. Fluorescence analysis revealed the close proximity of two different proteins on the MNPs. Enzyme-linked immunosorbent assay analysis showed that stoichiometrically equivalent amounts of the proteins were immobilized on the MNPs. Next, endoglucanase (EG) and β-glucosidase (BG) were immobilized on MNPs to give EG/BG-MNPs. The resulting MNPs were applied for the hydrolysis of celluloses, with rapid hydrolysis of carboxymethyl cellulose being observed. Furthermore, the fusion of the cellulose-binding domain to EG/BG-MNPs promoted improved hydrolysis activity against the insoluble cellulose. We could therefore conclude that the magnetosome display system can expand the possibilities of mimicking natural cellulosome organization on MNPs.
Microalgae are promising producers of biofuel due to higher accumulation of triacylglycerol (TAG). However, further improvement of the lipid metabolism is critical for feasible application of ...microalgae in industrial production of biofuel. Suppression of lipid degradation pathways is a promising way to remarkably increase the lipid production in model diatoms. In this study, we established an antisense-based knockdown (KD) technique in the marine oleaginous diatom, Fistulifera solaris. This species has a capability to accumulate high content of lipids. Tgl1 KD showed positive impact on cell growth and lipid accumulation in conventional culture in f/2 medium, resulting in higher oil contents compared to wild type strain. However, these impacts of Tgl1 KD were slight when the cells were subjected to the two-stage growth system. The Tgl1 KD resulted in slight change of fatty acid composition; increasing in C14:0, C16:0 and C16:1, and decreasing in C20:5. This study indicates that, although Tgl1 played a certain role in lipid degradation in F. solaris, suppression of only a single type of TAG lipase was not significantly effective to improve the lipid production. Comprehensive understanding of the lipid catabolism in this microalga is essential to further improve the lipid production.
Biointerfaces are regions where biomolecules, cells, and organic materials are exposed to environmental media or come in contact with other biomaterials, cells, and inorganic/organic materials. In ...this review article, six research topics on biointerfaces are described to show examples of state-of-art research approaches. First, biointerface design of nanoparticles for molecular detection is described. Functionalized gold nanoparticles can be used for sensitive detection of various target molecules, including chemical compounds and biomolecules, such as DNA, proteins, cells, and viruses. Second, the interaction between bacterial cell surfaces and material surfaces, including the introduction of advances in analytical methods and theoretical calculations, are explained as well as their applications to bioprocesses. Third, bioconjugation technologies for localizing functional proteins at biointerfaces are introduced, in particular, by focusing the potential of enzymes as a catalytic tool for designing different types of bioconjugates that function at biointerfaces. Forth topics is focusing on lipid–protein interaction in cell membranes as natural biointerfaces. Examples of membrane lipid engineering are introduced, and it is mentioned how their compositional profiles affect membrane protein functions. Fifth topic is the physical method for molecular delivery across the biointerface being developed currently, such as highly efficient nanoinjection, electroporation, and nanoneedle devices, in which the key is how to perforate the cell membrane. Final topic is the chemical design of lipid- or polymer-based RNA delivery carriers and their behavior on the cell interface, which are currently attracting attention as RNA vaccine technologies targeting COVID-19. Finally, future directions of biointerface studies are presented.
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•Marine diatom Fistulifera solaris JPCC DA0580as an alternative producer of PUFAs.•The maximal biomass productivity of 1.32±0.13g/(L·day).•The maximal EPA productivity was ...135.7±10.0mg/(L⋅day).•This is the highest productivity of PUFA in microalgae.
Polyunsaturated fatty acids (PUFAs), including eicosapentaenoic acid (EPA), have attracted attention owing to their health benefits for humans, as well as their importance in aquaculture and animal husbandry. Establishing a sustainable PUFA supply based on fish oils has been difficult due to their increasing demand. Therefore, alternative sources of PUFAs are required. In this research, we examined the potential of the marine oleaginous diatom Fistulifera solaris as an alternative producer of PUFAs. Optimization of culture conditions was carried out for high cell density cultivation, and a maximal biomass productivity of 1.32±0.13g/(L·day) was achieved. By slightly adjusting the culture conditions for EPA production, the maximal EPA productivity reached 135.7±10.0mg/(L·day). To the best of our knowledge, this is the highest EPA productivity among microalgae cultured under photoautotrophic conditions. This result indicates that F. solaris is a promising candidate host for sustainable PUFA production.
Oleaginous photosynthetic organisms such as microalgae are promising sources for biofuel production through the generation of carbon-neutral sustainable energy. However, the metabolic mechanisms ...driving high-rate lipid production in these oleaginous organisms remain unclear, thus impeding efforts to improve productivity through genetic modifications. We analyzed the genome and transcriptome of the oleaginous diatom Fistulifera solaris JPCC DA0580. Next-generation sequencing technology provided evidence of an allodiploid genome structure, suggesting unorthodox molecular evolutionary and genetic regulatory systems for reinforcing metabolic efficiencies. Although major metabolic pathways were shared with nonoleaginous diatoms, transcriptome analysis revealed unique expression patterns, such as concomitant upregulation of fatty acid/triacylglycerol biosynthesis and fatty acid degradation (β-oxidation) in concert with ATP production. This peculiar pattern of gene expression may account for the simultaneous growth and oil accumulation phenotype and may inspire novel biofuel production technology based on this oleaginous microalga.
Biofuel production using microalgae is believed to have the advantage of continuous year-round production over crop plants, which have strong seasonality. However, actual year-round production of ...microalgal lipids using outdoor mass cultivation has rarely been demonstrated. In our previous study, it was demonstrated that the oleaginous diatom,
, was culturable in outdoor bioreactors from spring to autumn, whereas biomass and lipid production in winter failed because
did not grow below 15 °C. Therefore, another candidate strain that is culturable in winter is required. In this study, a cold-tolerant diatom,
sp. JPCC CTDA0820, was selected as a promising candidate for biofuel production in winter. Laboratory-scale characterization revealed that this diatom was culturable at temperatures as low as 10 °C. Subsequently,
(April-October) and
sp. JPCC CTDA0820 (November-March) were cultured in outdoor open-pond bioreactors, wherein year-round production of diatom lipids was successfully demonstrated. The maximal values of areal productivities of biomass and lipids reached to 9.79 and 1.80 g/(m² day) for
, and 8.62 and 0.92 g/(m² day) for
sp. JPCC CTDA0820, respectively. With the combined use of these two diatom species, stable year-round production of microalgal lipids became possible.
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