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Farah Zaaboul, Husnain Raza, Cao Chen, Yuanfa Liu
A microscopic image of a section of a peanut seed shows that oil bodies (OBs) are a small droplet of oil that is dispersed throughout the whole seed. The protein profile of peanut's OBs recovered using the aqueous extraction method at different pH was found to have 2 oleosin isoforms of 14 and 16 kDa. Moreover, OBs essential amino acids are 1.52 higher than those in the PPI. Oleic acid and linoleic acid are the major fatty acids in both cold press peanut oil and OBs regardless of pH. Tocopherol content went from 270...
December 11, 2017: Journal of Food Science
Wenqing Li, Jing Yang, Jingbo Cai, Hongyu Wang, Haishan Tian, Jian Huang, Weidong Qiang, Linbo Zhang, Haiyan Li, Xiaokun Li, Chao Jiang
Recombinant human fibroblast growth factor 10 (rhFGF-10) is frequently used to treat patients with skin injuries. It can also promote hair growth. However, the effective application of rhFGF-10 is limited because of its poor stability and transdermal absorption. In this study, polymerase chain reaction (PCR) and Southern blotting were used to identify transgenic safflowers carrying a gene encoding an oleosin-rhFGF-10 fusion protein. The size and structural integrity of oleosin-rhFGF-10 in oil bodies extracted from transgenic safflower seeds was characterized by polyacrylamide gel electrophoresis and western blotting...
October 18, 2017: International Journal of Molecular Sciences
Ming-Mei Liu, Bin Qi, Zheng-Xu Liu, Jin-Shun Zhan, Kang Zhan, Guo-Qi Zhao
The aim of this study was to optimize the conditions for the extraction of low-abundance proteins (LAPs) and the removal of abundant proteins (APs; β-conglycinin and glycinin) from soybean meal. Single factor and orthogonal experiments were designed to determine the effects of four factors (isopropanol concentration, total extraction time, ultrasonic power, and ultrasonic time) on protein concentration in isopropanol extracts. Proteins in the isopropanol supernatant and the cold acetone precipitate of isopropanol were identified by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF-MS)...
2017: Journal of Zhejiang University. Science. B
Sehrish Manan, Muhammad Z Ahmad, Gaoyang Zhang, Beibei Chen, Basir U Haq, Jihong Yang, Jian Zhao
Soybean is an important oilseed crop and major dietary protein resource, yet the molecular processes and regulatory mechanisms involved in biosynthesis of seed storage substances are not fully understood. The B3 domain transcription factor (TF) LEC2 essentially regulates embryo development and seed maturation in other plants, but is not functionally characterized in soybean. Here, we characterize the function of a soybean LEC2 homolog, GmLEC2a, in regulating carbohydrate catabolism, triacylglycerol (TAG) biosynthesis, and seed development...
2017: Frontiers in Plant Science
Jing Yang, Weidong Qiang, Suping Ren, Shanyong Yi, Jian Li, Lili Guan, Linna Du, Yongxin Guo, Huilong Hu, Haiyan Li, Xiaokun Li
Basic fibroblast growth factor (bFGF) is a member of the fibroblast growth factors family. It is a highly specific mitogenic factor for many cell types, as though it be involved in wound repair, angiogenesis, nerve nutrition and embryonic development etc. Oil bodies have been applied for medicine, foodstuff and industry field. The heterogonous proteins expressed in oil bodies have distinct advantages, such as less purification steps and low costs. In this study, bFGF was expressed in A. thaliana seeds using oleosin fusion technology...
September 29, 2017: Gene
Yeming Chen, Yajing Chen, Luping Zhao, Xiangzhen Kong, Zhaoqi Yang, Yufei Hua
Peanut seeds are rich in oil, which exists as oil bodies (OBs). By extraction, peanut crude OBs are obtained and can be used as a food ingredient. In a previous study, it was found that the crude OBs contained an unknown protease, which hydrolyzed the oleosins. This would disrupt the integrity of OBs, and therefore, affect their physical and oxidative stability. In this study, the protein composition of crude OBs and some properties of the unknown protease were examined. The results showed that the protease was a two-chain (32 and 9kDa) aspartic protease, which showed high affinity for OBs...
February 15, 2018: Food Chemistry
Pascale Jolivet, Laure Aymé, Alexandre Giuliani, Frank Wien, Thierry Chardot, Yann Gohon
Lipid droplets are the major stock of lipids in oleaginous plant seeds. Despite their economic importance for oil production and biotechnological issues (biofuels, lubricants and plasticizers), numerous questions about their formation, structure and regulation are still unresolved. To determine water accessible domains of protein coating at lipid droplets surface, a structural proteomic approach has been performed. This technique relies on the millisecond timescale production of hydroxyl radicals by the radiolysis of water using Synchrotron X-ray white beam...
October 3, 2017: Journal of Proteomics
Buntora Pasaribu, Miki M C Wang, Pei-Luen Jiang
OBJECTIVE: To investigate the oil body protein and function in seeds of mature seagrass, Thalassia hemprichii. RESULTS: Seeds of mature seagrass T. hemprichii when stained with a fluorescent probe BODIPY showed the presence of oil bodies in intracellular cells. Triacylglycerol was the major lipid class in the seeds. Protein extracted from seagrass seeds was subjected to immunological cross-recognition with land plant seed oil body proteins, such as oleosin and caleosin, resulting in no cross-reactivity...
November 2017: Biotechnology Letters
Uta Jappe, Christian Schwager
PURPOSE OF REVIEW: The purpose of this review is to provide available data on a new class of allergens, the oleosins, and their diagnostic value. There is evidence that allergen extracts used for in vivo as well as in vitro diagnostic tests do not contain oleosins because these proteins are lipophilic and nearly insoluble in saline or aqueous solutions. So far, only oleosins of peanut, sesame and hazelnut have been registered as allergens. Reports on IgE-binding tests performed with oleosins of different species with sera from allergic patients show that IgE specific for oleosins are associated with severe allergic reactions which is why they should be part of the diagnostic tests in the future...
August 9, 2017: Current Allergy and Asthma Reports
Tzyy-Rong Jinn, Sheng-Kuo Hsieh, Yu-Jen Yu, Nou-Ying Tang, Jhao-Ren Lin
Here, we successfully use the oleosin-based fusion expression strategy coupled with the artificial oil body (AOB)-cyanogen bromide (CNBr) platform to produce bioactive MPB peptide which, in a manner identical to that of its native counterpart. In this study, the MPB peptide was interlinked with recombinant oleosin (rOle(127M→L)) by a methionine residue, a CNBr cleavage site. The recombinant rOle(127M→L)-MPB fusion gene was cloned into the pET30a(+) vector and was expressed in E. coli (DE3) after IPTG induction under optimized conditions...
July 24, 2017: Protein and Peptide Letters
Nodumo Nokulunga Zulu, Jennifer Popko, Krzysztof Zienkiewicz, Pablo Tarazona, Cornelia Herrfurth, Ivo Feussner
BACKGROUND: Microalgae are promising alternate and renewable sources for producing valuable products such as biofuel and essential fatty acids. Although this is the case, there are still challenges impeding on the effective commercial production of microalgal products. For instance, their product yield is still too low. Therefore, this study was oriented towards enhancing triacylglycerol (TAG) accumulation in the diatom Phaeodactylum tricornutum (strain Pt4). To achieve this, a type 2 acyl-CoA:diacylglycerol acyltransferase from yeast (ScDGA1) and the lipid droplet (LD) stabilizing oleosin protein 3 from Arabidopsis thaliana (AtOLEO3) were expressed in Pt4...
2017: Biotechnology for Biofuels
Luping Zhao, Xiangzhen Kong, Caimeng Zhang, Yufei Hua, Yeming Chen
P34 probable thiol protease (P34) and Gly m Bd 30K (30K) show high relationship with the protease of 24 kDa oleosin of soybean oil bodies. In this study, 9 day germinated soybean was used to separate bioprocessed P34 (P32) from bioprocessed 30K (28K). Interestingly, P32 existed as dimer, whereas 28K existed as monomer; a P32-rich sample had proteolytic activity and high cleavage site specificity (Lys-Thr of 24 kDa oleosin), whereas a 28K-rich sample showed low proteolytic activity; the P32-rich sample contained one thiol protease...
July 19, 2017: Journal of Agricultural and Food Chemistry
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...
August 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 present 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
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