Search results for: leaf protein concentrates

Authors: Shayli Varasteh Moradi, Wayne A. Johnston, Dejan Gagoski, Kirill Alexandrov

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The demand for a rapid and more efficient technique to identify protein-protein interaction particularly in the areas of therapeutics and diagnostics development is growing. The method described here is a rapid in vitro protein-protein interaction analysis approach based on AlphaLISA technology combined with Leishmania tarentolae cell-free protein production (LTE) system. Cell-free protein synthesis allows the rapid production of recombinant proteins in a multiplexed format. Among available in vitro expression systems, LTE offers several advantages over other eukaryotic cell-free systems. It is based on a fast growing fermentable organism that is inexpensive in cultivation and lysate production. High integrity of proteins produced in this system and the ability to co-express multiple proteins makes it a desirable method for screening protein interactions. Following the translation of protein pairs in LTE system, the physical interaction between proteins of interests is analysed by AlphaLISA assay. The assay is performed using unpurified in vitro translation reaction and therefore can be readily multiplexed. This approach can be used in various research applications such as epitope mapping, antigen-antibody analysis and protein interaction network mapping. The intra-viral protein interaction network of Zika virus was studied using the developed technique. The viral proteins were co-expressed pair-wise in LTE and all possible interactions among viral proteins were tested using AlphaLISA. The assay resulted to the identification of 54 intra-viral protein-protein interactions from which 19 binary interactions were found to be novel. The presented technique provides a powerful tool for rapid analysis of protein-protein interaction with high sensitivity and throughput.

Keywords: AlphaLISA technology, cell-free protein expression, epitope mapping, Leishmania tarentolae, protein-protein interaction

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Authors: Thi Thao Ninh, Yuri Trusov, José Ramón Botella

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Heterotrimeric G proteins, comprised of three subunits, α, β and γ, are involved in signal transduction pathways that mediate a vast number of processes across the eukaryotic kingdom. 23 Gα subunits are present in humans whereas most plant genomes encode for only one canonical Gα. The disparity observed between Arabidopsis, rice, and maize Gα-deficient mutant phenotypes suggest that Gα functions have diversified between eudicots and monocots during evolution. Alternatively, since the only Gα mutations available in dicots have been produced in Arabidopsis, the possibility exists that this species might be an exception to the rule. In order to test this hypothesis, we studied the G protein α subunit (TGA1) in tomato. Four tga1 knockout lines were generated in tomato cultivar Moneymaker using CRISPR/Cas9. The tga1 mutants exhibit a number of auxin-related phenotypes including changes in leaf shape, reduced plant height, fruit size and number of seeds per fruit. In addition, tga1 mutants have increased sensitivity to abscisic acid during seed germination, reduced sensitivity to exogenous auxin during adventitious root formation from cotyledons and excised hypocotyl explants. Our results suggest that Gα mutant phenotypes in tomato are very similar to those observed in monocots, i.e. rice and maize, and cast doubts about the validity of using Arabidopsis as a model system for plant G protein studies.

Keywords: auxin-related phenotypes, CRISPR/Cas9, G protein α subunit, heterotrimeric G proteins, tomato

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Authors: Indu Gaur, Neha Bhadauria, Shilpi Shilpi, Susmita Goswami, Prem D. Sharma, Prabir K. Paul

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The concept of organic agriculture has been accepted as novelty in Indian society, but there is no data available on the human pathogens colonizing plant parts due to such practices. Also, the pattern and mechanism of their colonization need to be understood in order to devise possible strategies for their prevention. In the present study, human pathogenic bacteria were isolated from organically grown tomato plants and five of them were identified as Klebsiella pneumoniae, Enterobacter ludwigii, Serratia fonticola, Stenotrophomonas maltophilia and Chryseobacterium jejuense. Tomato plants were grown in controlled aseptic conditions with 25±1˚C, 70% humidity and 12 hour L/D photoperiod. Six weeks old plants were divided into 6 groups of 25 plants each and treated as follows: Group 1: K. pneumonia, Group 2: E. ludwigii, Group 3: S. fonticola, Group 4: S. maltophilia, Group 5: C. jejuense, Group 6: Sterile distilled water (control). The inoculums for all treatments were prepared by overnight growth with uniform concentration of 108 cells/ml. Leaf samples from above groups were collected at 0.5, 2, 4, 6 and 24 hours post inoculation for the colony forming unit counts (CFU/cm2 of leaf area) of individual pathogens using leaf impression method. These CFU counts were used for the in vivo colonization assay and adherence assay of individual pathogens. Also, resistance of these pathogens to at least 12 antibiotics was studied. Based on these findings S. fonticola was found to be most prominently colonizing the phylloplane of tomato and was further studied. Tomato plants grown in controlled aseptic conditions same as mentioned above were divided into 2 groups of 25 plants each and treated as follows: Group 1: S. fonticola, Group 2: Sterile distilled water (control). Leaf samples from above groups were collected at 0, 24, 48, 72 and 96 hours post inoculation and homogenized in suitable buffers for surface and cell wall protein isolation. Protein samples thus obtained were subjected to isocratic SDS-gel electrophoresis and analyzed. It was observed that presence of S. fonticola could induce the expression of at least 3 additional cell wall proteins at different time intervals. Surface proteins also showed variation in the expression pattern at different sampling intervals. Further identification of these proteins by MALDI-MS and bioinformatics tools revealed the gene(s) involved in the interaction of S. fonticola with tomato phylloplane.

Keywords: cell wall proteins, human pathogenic bacteria, phylloplane, solanum lycopersicum

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Authors: Danutė Jablonskytė-Raščė, Vidas Damanauskas

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The wheat (Triticum) genus is divided into many species, of which only two are widely distributed in the world - common wheat (Triticum aestivum L.) and durum wheat (Triticum durum Desf.). Common (soft) wheat is the most common type of wheat in the world and the most suitable for the harsh climate of Lithuania, but the grains have lower protein content and poorer nutritional properties. Durum wheat is characterized by a high protein content of the grain, but it is a crop of warmer climates grown in southern countries, Italy, Spain, the United States, Egypt, etc. Today's important issue is food, its resources and quality. The research focuses on healthier food grown in our conditions, the quality of which recently depends a lot not only on the cultivation technology but also on the warming climate conditions. Climatic conditions change the distribution of fungi and their hosts. Plants that have grown in our climate for many years have adapted to the use of fungicides, so the aim is to study cereal varieties grown in warmer climates and compare them with our country's varieties, studying their nutritional value and the spread of fungal diseases. The field experiments of different varieties of wheat were conducted at Joniškėlis Experimental Station of the Lithuanian Research Centre for Agriculture and Forestry in 2023. The soil of the experimental site was Endocalcari-Endohypogleyic Cambisol (CMg-n-w-can). The research was designed to identify the resistance to leaf diseases and the nutritional value of various wheat varieties. This research aims to focus on healthier food grown in our conditions, the quality of which recently depends a lot not only on the cultivation technology but also on the conditions of the warming climate. The study found that hot and humid summer weather led to the spread of foliar diseases in wheat. Tan spot (Pyrenophora tritici-repentis) is mostly spread in wheat crops. This disease had an average prevalence of 86.90%. The wheat crop was sparse, so this year was unfavorable for the spread of powdery mildew (Blumeria graminis). Dry weather prevailed during the period of flowering of cereals, which prevented the spread of ear diseases. Examining the qualitative indicators of grain, it was found that durum wheat had the best parameters.

Keywords: varieties, wheat, leaf disease, grain quality

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Authors: Arfan Ali, Idrees Ahmad Nasir

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The critical evaluation of tolerance in tomato plants against the induced saline soil were assessed by transcript analysis of genes coding for products potentially involved in stress tolerance. A reverse transcriptase PCR experiment was performed with Hsp90-1, MT2, and GR1like protein genes using RNA isolated from different tissues of tomato plants. Four strains of Bacillus magisterium were inoculated with 100 Mm & 200 Mm concentrations of salt. Eleven treatments each ten replica pots were installed in green house experiment and the parameters taken into account were morphological (length, weight, number of leaves, leaf surface area), chemical (anthocyanin, chlorophyll-a, chlorophyll-b, carotenoids) and biological (gene expression). Results bare a response i.e. highest response of MT2 like gene was at 24 hpi and the highest levels of GR1 like protein transcript accumulation were detected at 36 hpi. The chemical and morphological parameters at diverse salt concentrations bequeath superlative response amongst strains which candidly flank on Zm7 and Zm4. Therefore, Bacillus magisterium Zm7 strains and somehow Zm4 strain can be used in saline condition to make plants tolerant. The overall performance of strains Zm7, Zm6, and Zm4 was found better for all studied traits under salt stress conditions. Significant correlations among traits root length, shoot length, number of leaves, leaf surface area, carotenoids, anthocyanin, chlorophyll-a and chlorophyll-b were found and suggested that the salt tolerance in tomato may be improved through the use of PGPR strains.

Keywords: Bacillus magisterium, gene expression glutathione reductase, metallothionein, PGPR, Rhizobacteria, saline

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Authors: Sudip Ghosh, Dipanwita Roy, Dipan Chatterjee, Paramita Bhattacharjee, Satadal Das

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Highest yield of eugenol-rich fractions from Cinnamomum tamala (bay leaf) leaves were obtained by supercritical carbon dioxide (SC-CO2), compared to hydro-distillation, organic solvents, liquid CO2 and subcritical CO2 extractions. Optimization of SC-CO2 extraction parameters was carried out to obtain an extract with maximum eugenol content. This was achieved using a sample size of 10 g at 55°C, 512 bar after 60 min at a flow rate of 25.0 cm3/sof gaseous CO2. This extract has the best combination of phytochemical properties such as phenolic content (1.77 mg gallic acid/g dry bay leaf), reducing power (0.80 mg BHT/g dry bay leaf), antioxidant activity (IC50 of 0.20 mg/ml) and anti-inflammatory potency (IC50 of 1.89 mg/ml). Identification of compounds in this extract was performed by GC-MS analysis and its antimicrobial potency was also evaluated. The MIC values against E. coli, P. aeruginosa and S. aureus were 0.5, 0.25 and 0.5 mg/ml, respectively.

Keywords: antimicrobial potency, Cinnamomum tamala, eugenol, supercritical carbon dioxide extraction

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Authors: F. A. M. Safwan, H. N. N. Dilrukshi, P. U. S. Peiris

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Less availability of high quality green forages leads to low productivity of national dairy herd of Sri Lanka. Growing grass and fodder to suit the production system is an efficient and economical solution for this problem. CO-3 is placed in a higher category, especially on tillering capacity, green forage yield, regeneration capacity, leaf to stem ratio, high crude protein content, resistance to pests and diseases and free from adverse factors along with other fodder varieties grown within the country. An experiment was designed to determine the effect of organic sheep manure, inorganic fertilizers and liquid sheep manure on growth, yield and nutritive value of CO-3. The study was consisted with three treatments; sheep manure (T1), recommended inorganic fertilizers (T2) and liquid sheep manure (T3) which was prepared using bucket fermentation method and each treatment was consisted with three replicates and those were assigned randomly. First harvest was obtained after 40 days of plant establishment and number of leaves (NL), leaf area (LA), tillering capacity (TC), fresh weight (FW) and dry weight (DW) were recorded and second harvest was obtained after 30 days of first harvest and same set of data were recorded. SPSS 16 software was used for data analysis. For proximate analysis AOAC, 2000 standard methods were used. Results revealed that the plants treated with T1 recorded highest NL, LA, TC, FW and DW and were statistically significant at first and second harvest of CO-3 (p˂ 0.05) and it was found that T1 was statistically significant from T2 and T3. Although T3 was recorded higher than the T2 in almost all growth parameters; it was not statistically significant (p ˃0.05). In addition, the crude protein content was recorded highest in T1 with the value of 18.33±1.61 and was lowest in T2 with the value of 10.82±1.14 and was statistically significant (p˂ 0.05). Apart from this, other proximate composition crude fiber, crude fat, ash, moisture content and dry matter were not statistically significant between treatments (p ˃0.05). In accordance with the results, it was found that the organic fertilizer is the best fertilizer for CO-3 in terms of growth parameters and crude protein content.

Keywords: fertilizer, growth parameters, Hybrid Napier CO-3, proximate composition

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Authors: Jackson Parker Galvan, Wenxuan Guo

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Unmanned aerial vehicle (UAV) technology is being increasingly adopted in high-throughput plant phenotyping for applications in plant breeding and precision agriculture. Winter wheat is an important cover crop for reducing soil erosion and protecting the environment in the Southern High Plains. Efficiently quantifying plant leaf area and biomass provides critical information for producers to practice site-specific management of crop inputs, such as water and fertilizers. The objective of this study was to estimate wheat biomass and leaf area index using UAV images. This study was conducted in an irrigated field in Garza County, Texas. High-resolution images were acquired on three dates (February 18, March 25, and May 15th ) using a multispectral sensor onboard a Matrice 600 UAV. On each data of image acquisition, 10 random plant samples were collected and measured for biomass and leaf area. Images were stitched using Pix4D, and ArcGIS was applied to overlay sampling locations and derive data for sampling locations.

Keywords: precision agriculture, UAV plant phenotyping, biomass, leaf area index, winter wheat, southern high plains

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Authors: Duc Dong Do, Ngoc Ha Tran, Thanh Hai Dang, Cao Cuong Dang, Xuan Huan Hoang

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Global aligning two protein-protein interaction networks is an essentially important task in bioinformatics/computational biology field of study. It is a challenging and widely studied research topic in recent years. Accurately aligned networks allow us to identify functional modules of proteins and/ororthologous proteins from which unknown functions of a protein can be inferred. We here introduce a novel efficient heuristic global network alignment algorithm called FASTAn, including two phases: the first to construct an initial alignment and the second to improve such alignment by exerting a local optimization repeated procedure. The experimental results demonstrated that FASTAn outperformed the state-of-the-art global network alignment algorithm namely SPINAL in terms of both commonly used objective scores and the run-time.

Keywords: FASTAn, Heuristic algorithm, biological network alignment, protein-protein interaction networks

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Authors: Abrha Reta, Ajebu Nurfeta, Genet Mengistu, Mohammed Beyan

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Fodder trees such as Ziziphus spina-christi have the potential to enhance the utilization of natural grazing resources and also to mitigate seasonal feed shortages. The experiment was conducted with the objective of evaluating the effect of supplementing Ziziphus spina-christi leaf meal (ZSCLM) to natural pasture hay on feed intake, body weight gain, digestibility, and carcass characteristics of Tigray highland sheep. A randomized complete block design was employed with 5 blocks based on initial body weight, and sheep were randomly assigned to five treatments. Treatments were: 100g concentrate mix + ad libtum natural pasture hay (T1), T1+ 100g ZSCLM (T2), T1 + 200g ZSCLM (T3), T1 + 300g ZSCLM (T4), and T1 + 400g ZSCLM (T5) on dry matter (DM) basis. Dry matter intake was greater (P<0.05) in sheep on T5 compared to T3 and T1, while the total DM intake among T2, T4, and T5 were similar. Crude protein and metabolizable energy intake differed (P<0.05) among treatments with highest and lowest values in T5 and T1, respectively. Average daily gain was higher (P<0.05) in sheep kept on T2, T3, and T4 diets than T1. Higher (P<0.05) DM digestibility was found in T4 and T5 than T1. The highest (P<0.05) OM and CP digestibility was observed in sheep fed T3, T4, and T5 diets. Rib eye muscle area was higher (P<0.05) for T4 than T1 and T2. Dressing percentage was similar (P>0.05) among treatments. The current study indicated that supplementation of Tigray highland sheep with 200g air-dried Ziziphus spina-christi leaf meal leaves with 100g of concentrate mixture in their diet significantly increased feed intake and apparent digestibility, body weight gain, hot carcass weight, and rib eye muscle area by improving feed conversion efficiency.

Keywords: body weight, carcass, digestibility, and ziziphus spina-christi leaf meal

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Authors: Xiao Zhou, Jianlin Cheng

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A single amino acid mutation can have a significant impact on the stability of protein structure. Thus, the prediction of protein stability change induced by single site mutations is critical and useful for studying protein function and structure. Here, we presented a deep learning network with the dropout technique for predicting protein stability changes upon single amino acid substitution. While using only protein sequence as input, the overall prediction accuracy of the method on a standard benchmark is >85%, which is higher than existing sequence-based methods and is comparable to the methods that use not only protein sequence but also tertiary structure, pH value and temperature. The results demonstrate that deep learning is a promising technique for protein stability prediction. The good performance of this sequence-based method makes it a valuable tool for predicting the impact of mutations on most proteins whose experimental structures are not available. Both the downloadable software package and the user-friendly web server (DNpro) that implement the method for predicting protein stability changes induced by amino acid mutations are freely available for the community to use.

Keywords: bioinformatics, deep learning, protein stability prediction, biological data mining

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Authors: Duygu Çimen, Nilay Bereli, Adil Denizli

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In this study is to synthesis magnetic nanoparticles for purify protein C. For this aim, N-Methacryloyl-(L)-histidine methyl ester (MAH) containing 2-hydroxyethyl methacrylate (HEMA) based magnetic nanoparticles were synthesized by using micro-emulsion polymerization technique for templating protein C via metal chelation. The obtained nanoparticles were characterized with Fourier transform infrared spectroscopy (FTIR), transmission electron microscopy (TEM), zeta-size analysis and electron spin resonance (ESR) spectroscopy. After that, they were used for protein C purification from aqueous solution to evaluate/optimize the adsorption condition. Hereby, the effecting factors such as concentration, pH, ionic strength, temperature, and reusability were evaluated. As the last step, protein C was determined with sodium dodecyl sulfate-polyacrylamide gel electrophoresis.

Keywords: immobilized metal affinity chromatography (IMAC), magnetic nanoparticle, protein C, hydroxyethyl methacrylate (HEMA)

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Authors: Ahmed M. Hjazi, Bader M. Hjazi

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Protein 4.1 is a crucial component of complex interactions between the cytoskeleton and other junctional complex proteins. When the gene encoding this protein is altered, resulting in reduced expression, or when the protein is absent, the red cell undergoes a significant structural change. This research aims to achieve a deeper comprehension of the biochemical effects of red cell protein deficiency. A Tandem Mass Spectrometry Analysis (TMT-MS/MS) of patient cells lacking protein 4.1 compared to three healthy controls was achieved by the Proteomics Institute of the University of Bristol. The SDS-PAGE and Western blotting were utilized on the original patient sample and controls to partially confirm TMT MS/MS data analysis of the protein-4.1-deficient cells. Compared to healthy controls, protein levels in samples lacking protein 4.1 had a significantly higher concentration of proteins that probably originated from reticulocytes. This could occur if the patient has an elevated reticulocyte count. The increase in chaperone and reticulocyte-associated proteins was most notable in this study. This may result from elevated quantities of reticulocytes in patients with hereditary elliptocytosis.

Keywords: hereditary elliptocytosis, protein 4.1, red cells, tandem mass spectrometry data.

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Authors: Yusuf Ali Abdulle, Azhar Uddin Keerio

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Background: Protein elicitors play a key role in signaling or displaying plant defense mechanisms and emerging as vital tools for bio-control of insects. This study was aimed at the characterization of the novel protein elicitor isolated from entomopathogenic fungi Lecanicillium lecanii (V3) strain and its activity against Whitefly, Bemisia tabaci in cotton. The sequence of purified elicitor protein showed 100% similarity with hypothetical protein LEL_00878 [Cordyceps confragosa RCEF 1005], GenBank no (OAA81333.1). This novel protein elicitor has 253 amino acid residues and 762bp with a molecular mass of 29 kDa. The protein recombinant was expressed in Escherichia coli using pET‐28a (+) plasmid. Effects of purified novel protein elicitor on Bemisia tabaci were determined at three concentrations of protein (i.e., 58.32, 41.22, 35.41 μg mL⁻¹) on cotton plants and were exposed to newly molted adult B.tabaci. Bioassay results showed a significant effect of the exogenous application of novel protein elicitor on B. tabaci in cotton. In addition, the gene expression analysis found a significant up-regulation of the major genes associated with salicylic acid (SA) and jasmonic acid (JA) linked plant defense pathways in elicitor protein-treated plants. Our results suggested the potential application of a novel protein elicitor derived from Lecanicillium lecanii as a future bio-intensive controlling approach against the whitefly, Bemisia tabaci.

Keywords: resistance, Lecanicillium lecanii, secondary metabolites, whitefly

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Authors: Angela U. Makolo, Temitayo A. Olagunju

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The knowledge of signaling pathways is central to understanding the biological mechanisms of organisms since it has been identified that in eukaryotic organisms, the number of signaling pathways determines the number of ways the organism will react to external stimuli. Signaling pathways are studied using protein interaction networks constructed from protein-protein interaction data obtained using high throughput experimental procedures. However, these high throughput methods are known to produce very high rates of false positive and negative interactions. In order to construct a useful protein interaction network from this noisy data, computational methods are applied to validate the protein-protein interactions. In this study, a computational technique to identify signaling pathways from a protein interaction network constructed using validated protein-protein interaction data was designed. A weighted interaction graph of the Saccharomyces cerevisiae (Baker’s Yeast) organism using the proteins as the nodes and interactions between them as edges was constructed. The weights were obtained using Bayesian probabilistic network to estimate the posterior probability of interaction between two proteins given the gene expression measurement as biological evidence. Only interactions above a threshold were accepted for the network model. A pathway was formalized as a simple path in the interaction network from a starting protein and an ending protein of interest. We were able to identify some pathway segments, one of which is a segment of the pathway that signals the start of the process of meiosis in S. cerevisiae.

Keywords: Bayesian networks, protein interaction networks, Saccharomyces cerevisiae, signalling pathways

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Authors: Kanlaya Kong-Ngern, Chutima Homwonk, Sittiruk Roytrakul

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In this research, we compared the proteomic profile of two rice leaf sheaths under salt stress, Thai moderately salt tolerant rice (Leaung Anan), and high salt tolerant rice (Pokkali). Seeds were grown in hydroponic culture for 21 days before NaCl was introduced initially at the level of 12 dS m⁻¹ for 10 days. Then the leaf sheath proteomes were analyzed by 1D-SDS-PAGE and NanoLC-MS/MS. In this study, 873 proteins were detected. Among these proteins, 219 proteins were known proteins and the other proteins were unnamed and unknown proteins. By using Mev software, we found that only 31 proteins in treated plants of both rice cultivars significantly expressed, 21 proteins were up-regulated and 10 proteins were down-regulated. Interestingly, the intensity of the 3 proteins in the Leaung Anan more expressed than in the Pokkali. The results indicate that the up-regulated proteins were more expressed in less tolerant rice may play an important role in helping rice to survive under salt stress.

Keywords: mass spectrometry, proteomics, rice leaf sheaths, salt stress

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Authors: Chinedu F. Anochie

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The presence of gangue particles in recovered metal concentrates has been a serious challenge to ore dressing engineers. Middlings lower the quality of concentrates, and in most cases, drastically affect the smelter terms, owing to exorbitant amounts paid by Mineral Processing industries as treatment charge. Models which encourage optimization of liberation operations have been utilized in most ore beneficiation industries to reduce the presence of locked particles in valuable concentrates. Moreover, methods such as incorporation of regrind mills, scavenger, rougher and cleaner cells, to the milling and flotation plants has been widely employed to tackle these concerns, and to optimize the grade–recovery relationship of metal concentrates. This work compared the crushing and grinding method of liberation, to the mine to mill route, by evaluating the proportion of middlings present in selectively processed complex Pb-Zn ore samples. To establish the effect of size reduction operations on the percentage of locked particles present in recovered concentrates, two similar samples of complex Pb- Zn ores were processed. Following blasting operation, the first ore sample was ground directly in a ball mill (Mine to Mill Route of Comminution), while the other sample was manually crushed, and subsequently ground in the ball mill (Crushing and Grinding Route of Comminution). The two samples were separately sieved in a mesh to obtain the desired representative particle sizes. An equal amount of each sample that would be processed in the flotation circuit was then obtained with the aid of a weighing balance. These weighed fine particles were simultaneously processed in the flotation circuit using the selective flotation technique. Sodium cyanide, Methyl isobutyl carbinol, Sodium ethyl xanthate, Copper sulphate, Sodium hydroxide, Lime and Isopropyl xanthate, were the reagents used to effect differential flotation of the two ore samples. Analysis and calculations showed that the degree of liberation obtained for the ore sample which went through the conventional crushing and grinding route of comminution, was higher than that of the directly milled run off mine (ROM) ore. Similarly, the proportion of middlings obtained from the separated galena (PbS) and sphalerite (ZnS) concentrates, were lower for the crushed and ground ore sample. A concise data which proved that the mine to mill method of size reduction is not the most ideal technique for the recovery of quality metal concentrates has been established.

Keywords: comminution, degree of liberation, middlings, mine to mill

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Authors: Gül Ebru Orhun

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A half diallel crossing design was carried out during 2011 and 2012 growing seasons under Çanakkale-Turkey ecological conditions. In this research, 20 F1 maize hybrids obtained by 6x6 half diallel crossing were used. Gene action for protein content and grain yield traits were explored in half set involving six elite inbred lines. According to the results diallel analysis dominance and additive gene variances were determined for protein content. Variance/Co-variance graphs revealed for grain yield and protein content traits. In this study, inheritance of grain yield and protein content demonstrated over-dominance type of gene action.

Keywords: protein, maize, inheritance, gene action

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Authors: Ayobami Adeyemo, Michael Chimonyo, Munyaradzi Marufu

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Gastrointestinal nematode (GNT) infection significantly hinder sustainable and profitable sheep production on rangelands. While vitamin E and protein supplementation have individually proven to improve host immunity to parasitism in lambs, to our knowledge, there is no information on the interaction of dietary vitamin E and protein supplementation on lamb growth and GIN faecal egg counts in naturally infected lambs. Therefore, the current study investigated the interaction of dietary protein and vitamin E supplementation on faecal egg counts (FEC) and growth performance of lambs. Twenty four Dohne Merino lambs aged 12 months were allocated equally to each of four treatment combinations, with six lambs in each treatment group for a period of eight weeks. Treatment one lambs received dietary protein and vitamin E (PE), treatment two lambs received dietary protein and no vitamin E (PNE), treatment three received dietary vitamin E and no protein (NPE), and treatment four received no dietary protein and vitamin E supplementation (NPNE). The lambs were allowed to graze on Pennisetum clandestinum contaminated with a heavy load of nematodes. Dietary protein supplementation increased (P < 0.01) average daily gain (ADG) and body condition scores (BCS). Dietary vitamin E supplementation had no effect (P > 0.05) on ADG and BCS. There was no interaction (P > 0.05) between dietary protein and vitamin E supplementation on ADG and BCS. Combined supplementation of dietary protein and vitamin E supplementation significantly reduced (P < 0.01) faecal egg counts and larval counts, respectively. Also, dietary protein and vitamin E supplementation reduced GNT faecal egg counts over the exposure period. The current findings support the hypothesis that the interaction of dietary protein and vitamin E supplementation reduced faecal egg counts and larval counts in lambs. This necessitates future findings on the interaction of dietary protein and vitamin E supplementation on blood associated profiles.

Keywords: gastrointestinal nematodes, nematode eggs, Haemonchus, Trichostrongylus

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Authors: Jin Choi, Zahra Aminikhoei, Yi-Oh Kim, Sang-Min Lee

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A 4 × 2 factorial experiment was conducted to determine the optimum dietary protein and lipid levels for juvenile fancy carp, Cyprinus carpio var. koi. Eight experimental diets were formulated to contain four protein levels (200, 300, 400, and 500 g kg-1) with two lipid levels (70 and 140 g kg-1). Triplicate groups of fish (initial weight, 12.1±0.2 g fish-1) were hand-fed the diets to apparent satiation for 8 weeks. Weight gain, daily feed intake, feed efficiency ratio and protein efficiency ratio were significantly (P < 0.0001) affected by dietary protein level, but not by dietary lipid level (P > 0.05). Weight gain and feed efficiency ratio tended to increase as dietary protein level increased up to 400 and 500 g kg-1, respectively. Daily feed intake of fish decreased with increasing dietary protein level and that of fish fed diet contained 500 g kg-1 protein was significantly lower than other fish groups. The protein efficiency ratio of fish fed 400 and 500 g kg-1 protein was lower than that of fish fed 200 and 300 g kg-1 protein. Moisture, crude protein and crude lipid contents of muscle and liver were significantly affected by dietary protein, but not by dietary lipid level (P > 0.05). The increase in dietary lipid level resulted in an increase in linoleic acid in liver and muscle paralleled with a decrease in n-3 highly unsaturated fatty acids content in muscle of fish. In considering these results, it was concluded that the diet containing 400 g kg-1 protein with 70 g kg-1 lipid level is optimal for growth and efficient feed utilization of juvenile fancy carp.

Keywords: fancy carp, dietary protein, dietary lipid, Cyprinus carpio, fatty acid

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Authors: Sajjad ur Rahman, Mariam Azam, Mukarram Bashir, Seemal Javaid, Aoun Muhammad, Muhammad Tahir, Jawad, Hannan Khan, Muhammad Zohaib

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Partially fermented soya hulls and wheat bran through Saccharomyces cerevisiae (DL-22 S/N) substantiated as a natural source for quality milk production. Saccharomyces cerevisiae (DL-22 S/N) were grown under in-vivo conditions and processed through two-step fermentation with substrates. The extra pure metabolites (XPM) were dried and processed for maintaining 1mm mesh size particles for supplementation of pelleted feed. Two groups of a cow (Holstein Friesian) having 8 animals of similar age and lactation were given the experimental concentrates. Group A was fed daily with 12gm of XPM and 22% protein-pelleted feed, while Group B was provided with no metabolites in their feed. In thirty-nine days of trial, improvement in the overall health, body score, milk protein, milk fat, ash, and solid not fat (SNF), yield, and incidence rate of mastitis was observed. The collected data revealed an improvement in milk production of 2.02 liter/h/d. However, a reduction (3.75%) in the milk fats and an increase in the milk SNF was around 0.58%. The ash content ranged between 6.4-7.5%. The incidence of mastitis was reduced to less than 2%.

Keywords: microbial metabolites, Saccharomyces cerevisiae, milk production, fermentation, post-biotic metabolites, immunity

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Authors: Muhammad H. Alu’datt, Inteaz Alli

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This study was conducted to prepare and evaluate the biological properties of protein co-precipitates from flaxseed and soybean. Protein was prepared by NaOH extraction through the mixing of soybean flour (Sf) and flaxseed flour (Ff) or mixtures of soybean extract (Se) and flaxseed extract (Fe). The protein co-precipitates were precipitated by isoelectric (IEP) and isoelectric-heating (IEPH) co-precipitation techniques. Effects of extraction and co-precipitation techniques on co-precipitate yield were investigated. Native-PAGE, SDS-PAGE were used to study the molecular characterization. Content and antioxidant activity of extracted free and bound phenolic compounds were evaluated for protein co-precipitates. Removal of free and bound phenolic compounds from protein co-precipitates showed little effects on the electrophoretic behavior of the proteins or the protein subunits of protein co-precipitates. Results showed that he highest protein contents and yield were obtained in for Sf-Ff/IEP co-precipitate with values of 53.28 and 25.58% respectively as compared to protein isolates and other co-precipitates. Results revealed that the Sf-Ff/IEP showed a higher content of bound phenolic compounds (53.49% from total phenolic content) as compared to free phenolic compounds (46.51% from total phenolic content). Antioxidant activities of extracted bound phenolic compounds with and without heat treatment from Sf-Ff/IEHP were higher as compared to free phenolic compounds extracted from other protein co-precipitates (29.68 and 22.84%, respectively).

Keywords: antioxidant, phenol, protein co-precipitate, yield

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Authors: Zhen Cao, Yu Zhu, Junxue Fu

Abstract:

Immunoassay, i.e., antigen-antibody reaction, is crucial for disease diagnostics. To achieve the adequate signal of the antigen protein detection, a large amount of sample and long incubation time is needed. However, the amount of protein is usually small at the early stage, which makes it difficult to detect. Unlike cells and DNAs, no valid chemical method exists for protein amplification. Thus, an alternative way to improve the signal is through particle manipulation techniques to concentrate proteins, among which dielectrophoresis (DEP) is an effective one. DEP is a technique that concentrates particles to the designated region through a force created by the gradient in a non-uniform electric field. Since DEP force is proportional to the cube of particle size and square of electric field gradient, it is relatively easy to capture larger particles such as cells. For smaller ones like proteins, a super high gradient is then required. In this work, three-dimensional Ag/SiO2 nanorods arrays, fabricated by an easy physical vapor deposition technique called as oblique angle deposition, have been integrated with a DEP device and created the field gradient as high as of 2.6×10²⁴ V²/m³. The nanorods based DEP device is able to enrich bovine serum albumin (BSA) protein by 1800-fold and the rate has reached 180-fold/s when only applying 5 V electric potential. Based on the above nanorods integrated DEP platform, an immunoassay of mouse immunoglobulin G (IgG) proteins has been performed. Briefly, specific antibodies are immobilized onto nanorods, then IgG proteins are concentrated and captured, and finally, the signal from fluorescence-labelled antibodies are detected. The limit of detection (LoD) is measured as 275.3 fg/mL (~1.8 fM), which is a 20,000-fold enhancement compared with identical assays performed on blank glass plates. Further, prostate-specific antigen (PSA), which is a cancer biomarker for diagnosis of prostate cancer after radical prostatectomy, is also quantified with a LoD as low as 2.6 pg/mL. The time to signal saturation has been significantly reduced to one minute. In summary, together with an easy nanorod fabrication and integration method, this nanorods based DEP platform has demonstrated highly sensitive immunoassay performance and thus poses great potentials in applications for early point-of-care diagnostics.

Keywords: dielectrophoresis, immunoassay, oblique angle deposition, protein concentration

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Authors: John Gichimu Mbaka, Jan Helmrich Martin von Baumbach, Celia Somlai, Denis Köpfer, Andreas Maeck, Andreas Lorke, Ralf Schäfer

Abstract:

Leaf litter decomposition is an important process providing energy to biotic communities. Additionally, methane gas (CH4) has been identified as an important alternative source of carbon and energy in some freshwater food webs.Flow regulation and dams can strongly alter freshwater ecosystems, but little is known about the effect of small impoundments on leaf litter decomposition and methane derived carbon in streams. In this study, we tested the effect of small water storage impoundments on leaf litter decomposition rates and methane derived carbon. Leaf litter decomposition rates were assessed by comparing treatment sites located close to nine impoundments (Rheinland Pfalz state, Germany) and reference sites located far away from the impoundments.CH4 concentrations were measured in eleven impoundments and correlated with the δ13C values of two subfamilies of chironomid larvae (i.e. Chironomini and Tanypodinae). Leaf litter break down rates were significantly lower in study sites located immediately above the impoundments, especially associated with a reduction in the abundance of shredders. Chironomini larvae had the lower mean δ13C values (‒29.2 to ‒25.5 ‰), than Tanypodinae larvae (‒26.9 to ‒25.3 ‰).No significant relationships were established between CH4 concentrations and δ13C values of chironomids (p> 0.05).Mean δ13C values of chironomid larvae (mean: ‒26.8‰, range: ‒ 29.2‰ to ‒ 25.3‰) were similar to those of sedimentary organic matter (SOM) (mean: ‒28.4‰, range: ‒ 29.3‰ to ‒ 27.1‰) and tree leaf litter (mean: ‒29.8 ‰, range: ‒ 30.5‰ to ‒ 29.1‰). In conclusion, this study demonstrates that small impoundments may have a negative effect on leaf litter decomposition in forest streams and that CH4 has limited influence on the benthic food web in stream impoundments.

Keywords: river functioning, chironomids, Alder tree, stable isotopes, methane oxidation, shredder

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Authors: Chahla Rahal, Philippe Refait

Abstract:

Oleuropein-rich extract from olive leaf and acid hydrolysates, rich in hydroxytyrosol and elenolic acid was prepared under different experimental conditions. These phenolic compounds may be used as a corrosion inhibitor. The inhibitive action of these extracts and its major constituents on the corrosion of copper in 0.5 M NaCl solution has been evaluated by potentiodynamic polarization, electrochemical impedance spectroscopy (EIS) and weight loss measurements. The product of extraction was analyzed with high performance liquid chromatography (HPLC), whose analysis shows that olive leaf extract are greatly rich in phenolic compounds, mainly Oleuropeine (OLE), Hydroxytyrosol (HT) and elenolic acid (EA). After the acid hydrolysis and high temperature of extraction, an increase in hydroxytyrosol concentration was detected, coupled with relatively low oleuropeine content and high concentration of elenolic acid. The potentiodynamic measurements have shown that this extract acts as a mixed-type corrosion inhibitor, and good inhibition efficiency is observed with the increase in HT and EA concentration. These results suggest that the inhibitive effect of olive leaf extract might be due to the adsorption of the various phenolic compounds onto the copper surface.

Keywords: olive leaf extract, oleuropein, voltammetry, copper, corrosion, HPLC, EIS

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Authors: M. K. Hossain, J. Keidel, O. Hensel, M. Diakite

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Fat reduced dairy products are holding a potential market due to health reason. Due to less creamy, and pleasantness, reduced and/or low-fat dairy products are getting less consumer acceptance whereas the fat molecule provides smooth, creamy and a pleasant mouthfeel in dairy products especially yogurt & ice cream. This study was aimed to investigate whether the application of microparticulated whey proteins (MWPs) processed by extrusion cooking, the reduced fat yogurt can achieve similar or higher creaminess compared to whole milk (3.8% fat) and skimmed milk (0.5% fat) yogurt. Full cream and skimmed milk were used to prepare natural stirred yogurt, as well as the dry matter content, also adjusted up to 16% with skimmed milk powder. Whey protein concentrates (WPC80) were used to produce MWPs in particle size of d50 > 5 µm, d50 3<5 µm and d50 < 3 µm through the hot-extrusion process with a screw speed of 400, 600 and 1000 rpm respectively. Furthermore, the commercially available microparticulated whey protein called Simplesse® was also applied in order to compare with extruded MWPs. The rheological and sensory properties of yogurt were assessed, and data were analyzed statistically. The applications of extruded MWPs with 600 and 1000 rpm were achieved significantly (p < 0.05) higher creaminess and preference compared to the whole and skimmed milk yogurt whereas, 400 rpm got lower preference. On the other hand, Simplesse® obtained the lowest creaminess and preference compared to other yogurts, although the contribution of dry matter in yogurt was same as extruded MWPs. The creaminess and viscosities were strongly (r = 0.62) correlated, furthermore, the viscosity from sensory evaluation and the dynamic viscosity of yogurt was also significantly (r = 0.72) correlated which clarifies that the performance of sensory panelists as well as the quality of the products.

Keywords: microparticulation, hot-extrusion, reduced-fat yogurt, whey protein concentrate

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Authors: Chahla Rahal, Philippe Refait

Abstract:

Oleuropein-rich extract from olive leaf and acid hydrolysates, rich in hydroxytyrosol and elenolic acid was prepared under different experimental conditions. These phenolic compounds may be used as a corrosion inhibitor. The inhibitive action of these extracts and its major constituents on the corrosion of copper in 0.5 M NaCl solution has been evaluated by potentiodynamic polarization, electrochemical impedance spectroscopy (EIS) and weight loss measurements. The product of extraction was analyzed with high performance liquid chromatography (HPLC), whose analysis shows that olive leaf extract are greatly rich in phenolic compounds, mainly Oleuropeine (OLE), Hydroxytyrosol (HT) and elenolic acid (EA). After the acid hydrolysis and high temperature of extraction, an increase in hydroxytyrosol concentration was detected, coupled with relatively low oleuropeine content and high concentration of elenolic acid. The potentiodynamic measurements have shown that this extract acts as a mixed-type corrosion inhibitor, and good inhibition efficiency is observed with the increase in HT and EA concentration. These results suggest that the inhibitive effect of olive leaf extract might be due to the adsorption of the various phenolic compounds onto the copper surface.

Keywords: Olive leaf extract, Oleuropein, hydroxytyrosol, elenolic acid , Copper, Corrosion, HPLC/DAD, Polarisation, EIS

Procedia PDF Downloads 228

Authors: Saleh H. Salmen, Sulaiman Ali Alharbi, Hany M. Yehia, Mohammad A. Khiyami, Milton Wainwright, Naiyf S. Alharbi, Arunachalam Chinnathambi

Abstract:

An antifungal protein synthesized by Bacillus cereus has been partially purified by the use of ammonium sulfate precipitation and Sephadex-G-200 column chromatography. The protein was produced from Bacillus cereus grown in potato Dextrose Broth Medium (PDB) at 30 ºC for 3 days at 100 rpm. The protein showed antagonistic effect against some fungi and yeasts. Crude extract from medium and semi-purified protein were tested in vitro against both fungi and yeasts using the disc diffusion method in order to detect the inhibitory effect of the protein. Zones of inhibition of the following diameter were found (mm) were Alternaria alternate (28), Rhodotorula glutinis (20), Fusarium sp. (16), Rhizopus sp. (15), Penicillium digitatum (13), Mucor sp. (13) and Aspergillus niger (10). The isolated protein was found to have a molecular weight of ~35kDa by sodium deodecyl sulfate-poly acrylamide gel electrophoresis. The data showed that the protein of Bacillus cereus has antifungal activity, a fact which points to the possibility of using it as a bio-control agent against some fungi, findings which emphasize the potential role of B. cereus as an important bio-control agent.

Keywords: bacillus cereus, ~35kDa protein, molds, yeasts

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Authors: M. N. Ofokansi, Onyemelukwe Chisom, Amauche Chukwuemeka, Ezema Onyinye

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The leaves of Clerodendrumcapitatum(Lamiaceae) is mostly used in the treatment of gastric ulcer in Nigerian folk medicine. The aim of this study was to evaluate the antiulcer activity of its crude methanol leaf extract and its ethyl acetate fraction in white albino rats. The effect of crude methanol leaf extract and its ethyl acetate fraction(250mg/kg, 500mg/kg) was evaluated using an absolute ethanol induced ulcer model. Crude methanol leaf extract and the ethyl acetate fraction was treated with distilled water and 6% Tween 80, respectively. crude methanol leaf extract was further investigated using a pylorus ligation induced ulcer model. Omeprazole was used as the standard treatment. Four groups of five albino rats of either sex were used. Parameters such as mean ulcer index and percentage ulcer protection were assessed in the ethanol-induced ulcer model, while the gastric volume, pH, and total acidity were assessed in the pyloric ligation induced ulcer model. Crude methanol leaf extract of Clerodendrumcapitatum(500mg/kg) showed a very highly significant reduction in mean ulcer index(p<0.001) in the absolute ethanol-induced model. ethyl acetate fraction of crude methanol leaf extract of Clerodendrumcapitatum(250mg/kg,500mg/kg) showed a very highly significant dose-dependent reduction in mean ulcer indices (p<0.001) in the absolute ethanol-induced model. The mean ulcer indices (1.6,2.2) with dose concentration (250mg/kg, 500mg/kg) of ethyl acetate fraction increased with ulcer protection (82.85%,76.42%) respectively when compared to the control group in the absolute ethanol-induced ulcer model. Crude methanol leaf extract of Clerodendrumcapitatum(250mg/kg, 500mg/kg) treated animals showed a highly significant dose-dependent reduction in mean ulcer index(p<0.01) with an increase in ulcer protection (56.77%,63.22%) respectively in pyloric ligated induced, ulcer model. Gastric parameters such as volume of gastric juice, pH, and total acidity were of no significance in the different doses of the crude methanol leaf extract when compared to the control group. The phytochemical investigation showed that the crude methanol leaf extracts Possess Saponins and Flavonoids while its ethyl acetate fraction possess only Flavonoids. The results of the study indicate that the crude methanol leaf extract and its ethyl acetate fraction is effective and has gastro protective and ulcer healing capacity. Ethyl acetate fraction is more potent than crude methanol leaf extract against ethanol-induced This result provides scientific evidence as a validation for its folkloric use in the treatment of gastric ulcer.

Keywords: gastroprotective, herbal medicine, anti-ulcer, pharmacology

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Authors: Katerina H. Takova, Ivan N. Minkov, Gergana G. Zahmanova

Abstract:

Molecular farming is the production of recombinant proteins in plants with the aim to use the protein as a purified product, crude extract or directly in the planta. Plants gain more attention as expression systems compared to other ones due to the cost effective production of pharmaceutically important proteins, appropriate post-translational modifications, assembly of complex proteins, absence of human pathogens to name a few. In addition, transient expression in plant leaves enables production of recombinant proteins within few weeks. Hepatitis E virus (HEV) is a causative agent of acute hepatitis. HEV causes epidemics in developing countries and is primarily transmitted through the fecal-oral route. Presently, all efforts for development of Hepatitis E vaccine are focused on the Open Read Frame 2 (ORF2) capsid protein as it contains epitopes that can induce neutralizing antibodies. For our purpose, we used the CMPV-based vector-pEAQ-HT for transient expression of HEV ORF2 in Nicotiana benthamina. Different molecular analysis (Western blot and ELISA) showed that HEV ORF2 capsid protein was expressed in plant tissue in high-yield up to 1g/kg of fresh leaf tissue. Electron microscopy showed that the capsid protein spontaneously assembled in low abundance virus-like particles (VLPs), which are highly immunogenic structures and suitable for vaccine development. The expressed protein was recognized by both human and swine HEV positive sera and can be used as a diagnostic reagent for the detection of HEV infection. Production of HEV capsid protein in plants is a promising technology for further HEV vaccine investigations. Here, we reported for a rapid high-yield transient expression of a recombinant protein in plants suitable for vaccine production as well as a diagnostic reagent. Acknowledgments -The authors’ research on HEV is supported with grants from the Project PlantaSYST under the Widening Program, H2020 as well as under the UK Biotechnological and Biological Sciences Research Council (BBSRC) Institute Strategic Programme Grant ‘Understanding and Exploiting Plant and Microbial Secondary Metabolism’ (BB/J004596/1). The authors want to thank Prof. George Lomonossoff (JIC, Norwich, UK) for his contribution.

Keywords: hepatitis E virus, plant molecular farming, transient expression, vaccines

Procedia PDF Downloads 128