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Chien Yu Huang, Anthony H C Huang
Plant cytosolic lipid droplets (LDs) are covered with a layer of phospholipids and oleosin and were extensively studied before those in mammals and yeast. Oleosin has short amphipathic N- and C-terminal peptides flanking a conserved 72-residue hydrophobic hairpin, which penetrates and stabilizes the LD. Oleosin is synthesized on endoplasmic reticulum (ER) and extracts ER-budding LDs to cytosol. To delineate the mechanism of oleosin targeting ER-LD, we have expressed modified-oleosin genes in Physcomitrella patens for transient expression and tobacco (Nicotiana tabacum) BY2 cells for stable transformation...
June 13, 2017: Plant Physiology
Sol Kim, Soo-Bin Lee, Chae-Seong Han, Mi-Na Lim, Sung-Eun Lee, In Sun Yoon, Yong-Sic Hwang
Oleosins are the most abundant proteins in the monolipid layer surrounding neutral storage lipids that form oil bodies in plants. Several lines of evidence indicate that they are physiologically important for the maintenance of oil body structure and for mobilization of the lipids stored inside. Rice has six oleosin genes in its genome, the expression of all of which was found to be responsive to abscisic acid (ABA) in our examination of mature embryo and aleurone tissues. The 5'-flanking region of OsOle5 was initially characterized for its responsiveness to ABA through a transient expression assay system using the protoplasts from suspension-cultured rice cells...
May 1, 2017: Journal of Plant Physiology
Jyun-Liang Lin, Jie Zhu, Ian Wheeldon
Eukaryotic biochemistry is organized throughout the cell in and on membrane-bound organelles. When engineering metabolic pathways this organization is often lost, resulting in flux imbalance and a loss of kinetic advantages from enzyme colocalization and substrate channeling. Here, we develop a protein-based scaffold for colocalizing multienzyme pathways on the membranes of intracellular lipid droplets. Scaffolds based on the plant lipid droplet protein oleosin and cohesin-dockerin interaction pairs recruited upstream enzymes in yeast ester biosynthesis to the native localization of the terminal reaction step, alcohol-O-acetyltransferase (Atf1)...
May 23, 2017: ACS Synthetic Biology
Wei Yan, Furong Li, Li Wang, Yaxin Zhu, Zhiyang Dong, Linhan Bai
A new gene encoding a lipase (designated as Lip-1) was identified from a metagenomic bacterial artificial chromosome(BAC) library prepared from a concentrated water sample collected from a hot spring field in Niujie, Eryuan of Yunnan province in China. The open reading frame of this gene encoded 622 amino acid residues. It was cloned, fused with the oleosin gene and over expressed in Escherichia coli to prepare immobilized lipase artificial oil body AOB-sole-lip-1. The monomeric Sole-lip-1 fusion protein presented a molecular mass of 102...
March 2017: Biotechnology Reports
A Baud, L Aymé, F Gonnet, I Salard, Y Gohon, P Jolivet, K Brodolin, P Da Silva, A Giuliani, B Sclavi, T Chardot, P Mercère, P Roblin, R Daniel
Synchrotron X-ray footprinting complements the techniques commonly used to define the structure of molecules such as crystallography, small-angle X-ray scattering and nuclear magnetic resonance. It is remarkably useful in probing the structure and interactions of proteins with lipids, nucleic acids or with other proteins in solution, often better reflecting the in vivo state dynamics. To date, most X-ray footprinting studies have been carried out at the National Synchrotron Light Source, USA, and at the European Synchrotron Radiation Facility in Grenoble, France...
May 1, 2017: Journal of Synchrotron Radiation
Christian Schwager, Skadi Kull, Jochen Behrends, Niels Röckendorf, Frauke Schocker, Andreas Frey, Arne Homann, Wolf-Meinhard Becker, Uta Jappe
BACKGROUND: Peanut allergy is one of the most common and most severe food allergies in Western countries and its accurate diagnosis to prevent potential life-threatening allergic reactions is crucial. However, aqueous extracts used for routine diagnostic measurements are devoid of lipophilic allergens such as oleosins. We have recently succeeded in the isolation and purification of these unique proteins, and the current study evaluates their allergenic potential and clinical relevance...
March 22, 2017: Journal of Allergy and Clinical Immunology
Tomoyuki Fujii
Coagulation and rheological behaviors of soy milk are reviewed from the viewpoint of colloidal dispersion system. From the results of relative viscosity in the range of small oil body volume fractions, oil bodies containing oleosin behave as rigid spheres. The Krieger-Dougherty equation was found to describe relative viscosities well under high oil body volume fraction. These results indicate that oil bodies in soy milk behave as though suspended matter. Cross-linking between colloid particles occurs when the coagulant is added, and bulky clusters are formed...
April 2017: Bioscience, Biotechnology, and Biochemistry
Tony Tumlirsch, Dieter Jendrossek
On the basis of bioinformatic evidence, we suspected that proteins with a CYTH (CyaB thiamine triphosphatase) domain and/or a CHAD (conserved histidine α-helical domain) motif might represent polyphosphate (polyP) granule-associated proteins. We found no evidence of polyP targeting by proteins with CYTH domains. In contrast, two CHAD motif-containing proteins from Ralstonia eutropha H16 (A0104 and B1017) that were expressed as fusions with enhanced yellow fluorescent protein (eYFP) colocalized with polyP granules...
April 1, 2017: Applied and Environmental Microbiology
Buntora Pasaribu, Chii-Shiarng Chen, Yue Ken Liao, Pei-Luen Jiang, Jason T C Tzen
Unique proteins including steroleosin, caleosin, oleosin-L, and oleosin-G have been identified in seed oil bodies of pine (Pinus massoniana). In this study, mature pollen grains with wing-like bladders were collected from pine (Pinus elliottii). Ultrastructural studies showed that oil bodies were present in pollen grains, but not the attached bladders, and the presence of oil bodies was further confirmed by fluorescent staining with BODIPY 493/503. Stable oil bodies were successfully purified from pine pollen grains, and analyzed to be mainly composed of triacylglycerols...
February 2017: Plant Physiology and Biochemistry: PPB
Zhenle Yang, Hongying Ji, Dantong Liu
Cyperus esculentus is unique in that it can accumulate rich oil in its tubers. However, the underlying mechanism of tuber oil biosynthesis is still unclear. Our transcriptional analyses of the pathways from pyruvate production up to triacylglycerol (TAG) accumulation in tubers revealed many distinct species-specific lipid expression patterns from oil seeds and fruits, indicating that in C. esculentus tuber: (i) carbon flux from sucrose toward plastid pyruvate could be produced mostly through the cytosolic glycolytic pathway; (ii) acetyl-CoA synthetase might be an important contributor to acetyl-CoA formation for plastid fatty acid biosynthesis; (iii) the expression pattern for stearoyl-ACP desaturase was associated with high oleic acid composition; (iv) it was most likely that endoplasmic reticulum (ER)-associated acyl-CoA synthetase played a significant role in the export of fatty acids between the plastid and ER; (v) lipid phosphate phosphatase (LPP)-δ was most probably related to the formation of the diacylglycerol (DAG) pool in the Kennedy pathway; and (vi) diacylglyceroltransacylase 2 (DGAT2) and phospholipid:diacylglycerolacyltransferase 1 (PDAT1) might play crucial roles in tuber oil biosynthesis...
December 2016: Plant & Cell Physiology
Yangyong Lv, Shuaibing Zhang, Jinshui Wang, Yuansen Hu
Wheat (Triticum aestivum L.) is an important crop worldwide. The physiological deterioration of seeds during storage and seed priming is closely associated with germination, and thus contributes to plant growth and subsequent grain yields. In this study, wheat seeds during different stages of artificial ageing (45°C; 50% relative humidity; 98%, 50%, 20%, and 1% Germination rates) and priming (hydro-priming treatment) were subjected to proteomics analysis through a proteomic approach based on the isobaric tandem mass tag labeling...
2016: PloS One
Abdulsamie Hanano, Ibrahem Almousally, Mouhnad Shaban, Farzana Rahman, Elizabeth Blee, Denis J Murphy
Contamination of aquatic environments with dioxins, the most toxic group of persistent organic pollutants (POPs), is a major ecological issue. Dioxins are highly lipophilic and bioaccumulate in fatty tissues of marine organisms used for seafood where they constitute a potential risk for human health. Lipid droplets (LDs) purified from date palm, Phoenix dactylifera, seeds were characterized and their capacity to extract dioxins from aquatic systems was assessed. The bioaffinity of date palm LDs toward 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), the most toxic congener of dioxins was determined...
2016: Frontiers in Plant Science
Qing Liu, Qigao Guo, Sehrish Akbar, Yao Zhi, Anna El Tahchy, Madeline Mitchell, Zhongyi Li, Pushkar Shrestha, Thomas Vanhercke, Jean-Philippe Ral, Guolu Liang, Ming-Bo Wang, Rosemary White, Philip Larkin, Surinder Singh, James Petrie
Potato tuber is a high yielding food crop known for its high levels of starch accumulation but only negligible levels of triacylglycerol (TAG). In this study, we evaluated the potential for lipid production in potato tubers by simultaneously introducing three transgenes, including WRINKLED 1 (WRI1), DIACYLGLYCEROL ACYLTRANSFERASE 1 (DGAT1) and OLEOSIN under the transcriptional control of tuber-specific (patatin) and constitutive (CaMV-35S) promoters. This coordinated metabolic engineering approach resulted in over a 100-fold increase in TAG accumulation to levels up to 3...
January 2017: Plant Biotechnology Journal
Chen Gao, Kevin B Vargo, Daniel A Hammer
Targeting nanoparticles for drug delivery has great potential for improving efficacy and reducing side effects from systemic toxicity. New developments in the assembly of materials afford the opportunity to expose cryptic targeting domains in tissue-specific microenvironments in which certain proteases are expressed. Here, recombinant proteins are designed to combine the responsiveness to environmental proteases with specific targeting. Materials made recombinantly allow complete control over amino acid sequence, in which each molecule is identically functionalized...
September 2016: Macromolecular Bioscience
Min Liu, Shenghui Chu, Jun Ai, Haiyan Li, Zhiyu Chen, Sisi Huang, Chao Jiang, Xiaokun Li
OBJECTIVE: To determine an efficient expression strategy in Arabidopsis thaliana and to determine whether a dimeric oleosin fusion approach could achieve keratinocyte growth factor-2 (KGF2) expression and bioactivity. RESULTS: Higher recombinant protein accumulation was observed in the two dimeric oleosin constructs than in a single-oleosin-fused protein (O::KGR2) or KGF2 control. Highest expression was in O-O::KGF2-transgenic seeds. MTT assay in FGFR2 III b-BaF3 cells revealed comparable levels of bioactivity due to O-O::KGF2 and O::KGR2, whereas O::KGF2-O had no effect on FGFR2 III b-BaF3 opithelial cell growth...
September 2016: Biotechnology Letters
Ming-Der Huang, Anthony H C Huang
Subcellular lipid droplets (LDs) in diverse plant cells and species are coated with stabilizing oleosins of at least five phylogenic lineages and perform different functions. We examined two types of inadequately studied LDs for coated oleosins and their characteristics. The epidermis but not mesophyll of leaves of vanilla (Vanilla planifolia) and most other Asparagales species contained solitary and clustered LDs (<0.5 μm), some previously studied by electron microscopy and speculated to be for cuticle formation...
July 2016: Plant Physiology
Li Zhang, Shi-Bo Wang, Qi-Gang Li, Jian Song, Yu-Qi Hao, Ling Zhou, Huan-Quan Zheng, Jim M Dunwell, Yuan-Ming Zhang
Seed oils provide a renewable source of food, biofuel and industrial raw materials that is important for humans. Although many genes and pathways for acyl-lipid metabolism have been identified, little is known about whether there is a specific mechanism for high-oil content in high-oil plants. Based on the distinct differences in seed oil content between four high-oil dicots (20~50%) and three low-oil grasses (<3%), comparative genome, transcriptome and differential expression analyses were used to investigate this mechanism...
2016: PloS One
Philip E Johnson, Rebekah L Sayers, Lee A Gethings, Anuradha Balasundaram, Justin T Marsh, James I Langridge, E N Clare Mills
Profiling allergens in complex food ingredients used in oral food challenges and immunotherapy is crucial for regulatory acceptance. Mass spectrometry based analysis employing data-independent acquisition coupled with ion mobility mass spectrometry-mass spectrometry (DIA-IM-MS) was used to investigate the allergen composition of raw peanuts and roasted peanut flour ingredients used in challenge meals. This comprehensive qualitative and quantitative analysis using label-free approaches identified and quantified 123 unique protein accessions...
June 7, 2016: Analytical Chemistry
Sweta Kumari, Lucia Joseph Memba, Anil Dahuja, T Vinutha, Supradip Saha, Archana Sachdev
Defatting soybean by sophisticated oil extraction method utilising supercritical CO2 resulted in a significant decrease in the residual phospholipids (PLs) compared with soymeal obtained by conventional cold percolation method utilising hexane as the extraction solvent. Interestingly, the levels of residual PLs showed a proportionate relationship with thiobarbituric acid (TBA) number, an indicator of lipid peroxidation responsible for off-flavour generation. Furthermore, two oleosins (18 and 24 kDa) were isolated from the oil bodies extracted from soybean seeds and positively characterised for phospholipase A2 (PLA2) activity, suggesting their plausible involvement in off-flavour generation in soymeal...
August 15, 2016: Food Chemistry
Malgorzata Teodorowicz, Rozine J Terlouw, Ad Jansen, Huub F J Savelkoul, Janneke Ruinemans-Koerts
BACKGROUND: Sesame seed is an allergen of growing importance worldwide. However, knowledge of the clinically relevant sesame allergen and its cross-reactivity with homologous allergens is limited. The aim of this study was the immunological characterization of Dutch sesame seed-allergic patients and evaluation of cross-reactivity between sesame seed, tree nut and pollen allergens using different sources of allergen extracts. METHODS: Six patients with a medical history of sesame seed allergy were included, i...
2016: International Archives of Allergy and Immunology
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