Search results for: vegetable protein (VP)

Authors: Vita Strazdina, Aleksandrs Jemeljanovs, Vita Sterna

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Not all proteins have the same nutritional value, since protein quality strongly depends on its amino acid composition and digestibility. The meat of game animals could be a high protein source because of its well-balanced essential amino acids composition. Investigations about biochemical composition of game meat such as wild boar (Sus scrofa scrofa), roe deer (Capreolus capreolus) and beaver (Castor fiber) are not very much. Therefore, the aim of the investigation was evaluate protein composition of game meat hunted in Latvia. The biochemical analysis, evaluation of connective tissue and essential amino acids in meat samples were done, the amino acids score were calculate. Results of analysis showed that protein content 20.88-22.05% of all types of meat samples is not different statistically. The content of connective tissue from 1.3% in roe deer till 1.5% in beaver meat allowed classified game animal as high quality meat. The sum of essential amino acids in game meat samples were determined 7.05–8.26g100g-1. Roe deer meat has highest protein content and lowest content of connective tissues among game meat hunted in Latvia. Concluded that amino acid score for limiting amino acids phenylalanine and tyrosine is high and shows high biological value of game meat.

Keywords: dietic product, game meat, amino acids, scores

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Authors: Mandana Hoseini

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During last decades, intensive selection for higher growth rate in broiler chickens has accelerated daily body weight gain, which this has changed/increased the trends and amounts of nutrient requirements in the diet. As a result, considerable studies have been focused on the better determination of protein/amino acids requirements in modern broiler diets. One approach to minimize dietary crude protein inclusion levels is substitution of some of the dietary crude protein with synthetic amino acids. In addition, using synthetic forms of limiting essential amino acids in the diet could help better coincidence of dietary protein with ideal protein concept, which this in turn, minimizes nitrogen dissipation and environmental pollution. Threonine is usually considered as the third limiting amino acid in broiler diets. Recent studies have been demonstrated that dietary supplemental threonine would optimize growth performance, immune system, intestinal morphology, as well as oxidative defense in broiler chickens. In this review, threonine metabolism and its effects in relation with different aspects of broiler performance have been discussed.

Keywords: immune system, intestine, performance, requirement, threonine

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Authors: M. Manjunath, R. Rakesh, Y. T. Krishne Gowda, G. Panduranga Murthy

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The availability of energy resources plays a vital role in the progress of a country. Over the last decades, there is an increase in the consumption of energy worldwide resulting in the depletion of fossil fuels. This necessitates dependency on other countries for energy resources. Therefore, a renewable eco-friendly alternate fuel is replaced in place of fossil fuel which can be vegetable oils as a substitute fuel for diesel. Since oils are more viscous it cannot be used directly in CI engines without any engine modification. Thus, a conversion of vegetable oils to biodiesel is done by a Transesterification process. The present paper is restricted to Biofuel substitute for diesel and which can be obtained from a number of edible and non-edible oil resources. The oil from these resources can be Transesterified by a suitable method depending on its FFA content for the production of biodiesel and that can be used to operate CI engine. In this work, an attempt is made to test the performance of CI engine using Transesterified peanut and sunflower oil methyl esters blends with diesel.

Keywords: SOME, POME, BMEP, BSFC, BTE

Procedia PDF Downloads 473

Authors: Sirikwan Ponprateep, Anchalee Tassanakajon, Vichien Rimphanitchayakit

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A Kazal-type serine proteinase inhibitor, SPIPm2, was abundantly expressed in the hemocytes and secreted into shrimp plasma has anti-viral property against white spot syndrome virus (WSSV). To discover the molecular mechanism of antiviral activity, the binding assay showed that SPIPm2 bind to the components of viral particle and shrimp hemocyte. From our previous report, viral target protein of SPIPm2 was identified, namely WSV477 using yeast two-hybrid screening. WSV477 is an early gene product of WSSV and involved in viral propagation. In this study, the co-immunoprecipitation technique and Tandem Mass Spectrometry (LC-MS/MS) was used to identify the target protein of SPIPm2 from shrimp hemocyte. The target protein of SPIPm2 was cyclophilin A. In vertebrate, cyclophilin A or peptidylprolyl isomerase A was reported to be the immune suppressor interacted with cyclosporin A involved in immune defense response. The recombinant cyclophilin A from Penaeus monodon (rPmCypA) was produced in E.coli system and purified using Ni-NTA column to confirm the protein-protein interaction. In vitro pull-down assay showed the interaction between rSPIPm2 and rPmCypA. To study the biological function of these proteins, the expression analysis of immune gene in shrimp defense pathways will be investigated after rPmCypA administration.

Keywords: cyclophilin A, protein-protein interaction, Kazal-type serine proteinase inhibitor, Penaeus monodon

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Authors: Waode Nurmayani, Nikmatul Riswanda

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Broilers are chickens which are kept with the most efficient time and hoped get a good body weight. All things are done, for example with the improvement of feed and use antibiotics. Feed cost is the most cost to be spent. Nearly 80% of the cost is spent just for buy feed. Earthworm (Lumbricus rubellus) is a good choice to reduce the cost of feed protein source. The Earthworm has a high crude protein content of about 48.5%-61.9%, rich with proline amino acid about 15% of the 62 amino acids. Not only about protein, this earthworm also has a role in disease prevention. Prevention of disease in livestock usual with use feed supplement. Earthworm (Lumbricus rubellus) is one of the natural materials used as feed. In addition, several types of earthworms that have been known to contain active substances about antibacterial pathogens namely Lumbricus rubellus. The earthworm could be used as an antibiotic because it contain the antibody of Lumbricine active substance. So that, this animal feed from Lumbricus rubellus could improve the performance of broilers. Bioactive of anti-bacterial is called Lumbricine able to inhibit the growth of pathogenic bacteria in the intestinal wall so that the population of pathogenic bacteria is reduced. The method of write in this scientific writing is divided into 3 techniques, namely data completion, data analysis, and thinking pan from various literature about earthworm (Lumbricus rubellus) as broiler feed. It is expected that innovation of feed material of earthworm (Lumbricus rubellus) could reduce the cost of protein feed and the use of chemical antibiotics.

Keywords: earthworm, broiler, protein, antibiotic

Procedia PDF Downloads 157

Authors: N. Mahmoudnia, M. Khormali

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This experiment was conducted to determine the apparent protein digestibility of poultry byproduct meal (PBPM) from two industrial poultry slaughter-houses on Ross 308 male broiler chickens in independent comparisons. The experiment consisted of seven dietary treatments and three replicates per treatment with three broiler chickens per replicate in a completely randomized design. Dietary treatments consisted of a control corn- soybean diet, and levels 3, 6, and 9% PBPM produced by slaughter-house 1 and levels 3, 6, and 9% PBPM produced by slaughter house 2. Chromic oxide was added to the experimental diets as an indigestible marker. The apparent protein digestibility of each diet were determined with two methods of sample collection of ileum and excreta in 21-28 d of age. The results this experiment showed that use of PBPM had no significant effect on the performance of broiler chicks during period of experiments. The apparent protein digestibility of PBPM groups was significantly higher than control group by excreta sampling procedure (P<0.05). Using of PBPM 2 significantly (P<0.05) decreased the apparent protein digestibility values based on ileum sampling procedure vs control (85.21 vs. 90.14).Based results of this experiment,it is possible to use of PBPM 1 in broiler chicken.

Keywords: poultry by-product meal, apparent protein digestibility, independed comparison, broiler chicken

Procedia PDF Downloads 492

Authors: Neway Adele, Adey Feleke

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Access to clean drinking water is a basic human right. However, an estimated 1.2 billion people across the world consume unclean water daily. Interest has been growing in natural coagulants as the health and environmental concerns of conventional chemical coagulants are rising. Natural coagulants have the potential to serve as alternative water treatment agents. In this study, Lupinus albus, Moringa stenopetala, Trigonella foenum-graecum and Vicia faba protein extracts were evaluated as natural coagulants for water treatment. The protein extracts were purified from crude extracts using a protein purifier, and protein concentrations were determined by the spectrophotometric method. Small-volume coagulation efficiency tests were conducted on raw water taken from the Legedadi water treatment plant. These were done using a completely randomized design (CRD) experiment with settling times of 0 min (initial time), 90 min, 180 min and 270 min and protein extract doses of 5 mg/L, 10 mg/L, 15 mg/L and 20 mg/L. Raw water as negative control and polyelectrolyte as positive control were also included. The optical density (OD) values were measured for all the samples. At 270 min and 20 mg/L, the coagulation efficiency percentages for Lupinus albus, Moringa stenopetala, Trigonella foenum-graecum and Vicia faba protein extracts were 71%, 89%, 12% and 67% in the water sample collected in April 2019 respectively. Similarly, Lupinus albus, Moringa stenopetala and Vicia faba achieved 17%, 92% and 12% at 270 min settling times and 5 mg/L, 20 mg/L and 10 mg/L concentration in the water sample collected from August 2019, respectively. Negative control (raw water) and polyelectrolyte (positive control) were also 6 − 10% and 89 − 94% at 270 min settling time in April and August 2019, respectively. Among the four protein extracts, Moringa stenopetala showed the highest coagulation efficiency, similar to polyelectrolyte. This study concluded that Moringa stenopetala protein extract could be used as a natural coagulant for water purification in both sampling times.

Keywords: coagulation efficiency, extraction, natural coagulant, protein extract

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Authors: Neelam Patel

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Farmers in peri-urban areas of developing countries depend on wastewater for Irrigation but with great environmental and health hazards. Since, irrigation with wastewater is growing in the developing countries but its suitability to environment and other health factors should be checked. Metal pollution is a very serious issue these days, various neuro, physical and mental disorders are prevailing due to the metal pollution. Waste water contaminated with heavy metals got accumulated in the soil and then bioaccumulated in the vegetables irrigated with waste water. A 3-year field experiment on cauliflower has been done by using wastewater with two different methods of irrigation i.e. Drip and Flood irrigation and checked the impact on the cauliflower and soil quality. Heavy metals (Cr, Cu, Ni, Zn and Pb) have been studied in wastewater used for the irrigation and their accumulation in the soil and vegetable was studied. The study reveals that the concentration of heavy metals increases by 100 times from initial in soil. After 3 years, the concentration of Copper(41 ppm) Chromium(39.4 ppm) Lead(62.2ppm) Zinc(100.5 ppm) and Nickel(75.7 ppm) in Flood irrigated soil while in Drip irrigated soil , Copper (36.4 ppm) Chromium(36.8 ppm) Lead(53.7 ppm) Zinc(70.3 ppm) and Nickel (53.9 ppm). In vegetable, the wastewater irrigated shows an increase in the concentration of metals with the time and the accumulation of Nickel (6.98ppm), Lead (30.18 ppm) and Zinc (55.83 ppm) in drip irrigated while in flood irrigated, Nickel (30.58 ppm), Lead (73.95ppm) Zinc (93.50 ppm) and Copper (54.58 ppm) in edible part of cauliflower which is above the permissible limits suggested by different international agencies. On other hand, the nutrients content i.e. Nitrogen, Phosphorus and Potassium in soil was increased in concentration with time. The study pointed out that the metal contaminated waste water consisting the nutrients in it but also heavy metals which causes health issues in human. While the increase in concentration of nutrients in the soil indirectly helpful to the farmers economically by restricting the use of fertilizers. But the metal pollution directly affects the health of human being. The different method of irrigation suggested that the drip irrigated vegetable acquired less metal then the flood one and is a better combo with the waste water for the irrigation.

Keywords: drip irrigation, heavy metals, metal contamination, waste water

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Authors: Anna Trusek-Holownia, Andrzej Noworyta

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Using an enzyme of known specificity the hydrolysis of protein was carried out in a controlled manner. The aim was to obtain oligopeptides being the so-called active peptides or their direct precursors. An original way of expression of the protein hydrolysis kinetics was introduced. Peptide bonds contained in the protein were recognized as a diverse-quality substrate for hydrolysis by the applied protease. This assumption was positively verified taking as an example the hydrolysis of albumin by thermolysin. Peptide linkages for this system should be divided into at least four groups. One of them is a group of bonds non-hydrolyzable by this enzyme. These that are broken are hydrolyzed at a rate that differs even by tens of thousands of times. Designated kinetic constants were k'F = 10991.4 L/g.h, k'M = 14.83L/g.h, k'S about 10-1 L/g.h for fast, medium and slow bonds, respectively. Moreover, a procedure for unfolding of the protein, conducive to the improved susceptibility to enzymatic hydrolysis (approximately three-fold increase in the rate) was proposed.

Keywords: peptide bond hydrolysis, kinetics, enzyme specificity, biologically active peptides

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Authors: Ovanes Chakoyan

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MOH (mineral oil hydrocarbons), which consist of mineral oil saturated hydrocarbons(MOSH) and mineral oil aromatic hydrocarbons(MOAH), are present in various products such as vegetable oils, animal feed, foods, and infant formula. Contamination of foods with mineral oil hydrocarbons, particularly mineral oil aromatic hydrocarbons(MOAH), exhibiting carcinogenic, mutagenic, and hormone-disruptive effects. Identifying toxic substances among the many thousands comprising mineral oils in food samples is a difficult analytical challenge. A method based on an offline-solid phase extraction approach coupled with gas chromatography-triple quadrupole(GC-MS/MS) was developed for the determination of MOSH/MOAH in various products such as vegetable oils, animal feed, foods, and infant formula. A glass solid phase extraction cartridge loaded with 7 g of activated silica gel impregnated with 10 % silver nitrate for removal of olefins and lipids. The MOSH/MOAH fractions were eluated with hexane and hexane: dichloromethane : toluene, respectively. Each eluate was concentrated to 50 µl in toluene and injected on splitless mode into GC-MS/MS. Accuracy of the method was estimated as measurement of recovery of spiked oil samples at 2.0, 15.0, and 30.0 mg kg -1, and recoveries varied from 85 to 105 %. The method was applied to the different types of samples (sunflower meal, chocolate ships, santa milk chocolate, biscuits, infant milk, cornflakes, refined sunflower oil, crude sunflower oil), detecting MOSH up to 56 mg/kg and MOAH up to 5 mg/kg. The limit of quantification(LOQ) of the proposed method was estimated at 0.5 mg/kg and 0.3 mg/kg for MOSH and MOAH, respectively.

Keywords: MOSH, MOAH, GC-MS/MS, foods, solid phase extraction

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Authors: Lynn Lehmann, Dinorah Leyva, Ana Lazic, Stefan Duhr, Philipp Baaske

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Characterization of biomolecular interactions, such as protein-protein, protein-nucleic acid or protein-small molecule, provides critical insights into cellular processes and is essential for the development of drug diagnostics and therapeutics. Here we present a novel, label-free, and tether-free technology to analyze picomolar to millimolar affinities of biomolecular interactions by Microscale Thermophoresis (MST). The entropy of the hydration shell surrounding molecules determines thermophoretic movement. MST exploits this principle by measuring interactions using optically generated temperature gradients. MST detects changes in the size, charge and hydration shell of molecules and measures biomolecule interactions under close-to-native conditions: immobilization-free and in bioliquids of choice, including cell lysates and blood serum. Thus, MST measures interactions under close-to-native conditions, and without laborious sample purification. We demonstrate how MST determines the picomolar affinities of antibody::antigen interactions, and protein::protein interactions measured from directly from cell lysates. MST assays are highly adaptable to fit to the diverse requirements of different and complex biomolecules. NanoTemper´s unique technology is ideal for studies requiring flexibility and sensitivity at the experimental scale, making MST suitable for basic research investigations and pharmaceutical applications.

Keywords: biochemistry, biophysics, molecular interactions, quantitative techniques

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Authors: B. F. Olanipekun, O. J. Oyelade, C. O. Osemobor

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Pigeon pea is a very good source of protein and micronutrient, but it is being underutilized in Nigeria because of several constraints. This research considered the effect of different processing methods on the quality attributes of pigeon pea used in bread production towards enhancing its utility. Pigeon pea was obtained at a local market and processed into the flour using three processing methods: soaking, sprouting and roasting and were used to bake bread in different proportions. Chemical composition and sensory attributes of the breads were thereafter determined. The highest values of protein and ash contents were obtained from 20 % substitution of sprouted pigeon pea in wheat flour and may be attributable to complex biochemical changes occurring during hydration, to invariably lead to protein constituent being broken down. Hydrolytic activities of the enzymes from the sprouted sample resulted in improvement in the constituent of total protein probably due to reduction in the carbohydrate content. Sensory qualities analyses showed that bread produced with soaked and roasted pigeon pea flours at 5 and 10% inclusion, respectively were mostly accepted than other blends, and products with sprouted pigeon pea flour were least accepted. The findings of this research suggest that supplementing wheat flour with sprouted pigeon peas have more nutritional potentials. However, with sensory analysis indices, the soaked and roasted pigeon peas up to 10% are majorly accepted, and also can improve the nutritional status. Overall, this will be very beneficial to population dependent on plant protein in order to combat malnutrition problems.

Keywords: pigeon pea, processing, protein, malnutrition

Procedia PDF Downloads 250

Authors: Maria Magdy Danial Riad

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The objective of this study was to investigate the effect of different protein sources with different degradability ratios during late gestation of ewes on colostrum composition and its IgG concentration, body weight change of dams, and birth weight of their lambs. Objectives: 35 multiparous native crossbred ewes (BW= 59±2.5kg) were randomly allocated to five dietary treatments (7 ewes / treatment) for 2 months prior to lambing. Methods: Experimental diets were isonitrogenous (12.27% CP) and isocaloric (2.22 Mcal ME/kg DM). In diet I (the control), solvent extract soybeans (SESM 33% RUP of CP), II feed grade urea (FGU 31% RUP), III slow release urea (SRU 31% RUP). As sources of undegradable protein, extruded expeller SBM-EESM 40 (37% RUP) and extruded expeller SBM-EESM 60 (41% RUP) were used in groups IV and V, respectively. Results showed no significant effect on feed intake, crude protein (CP), metabolizable energy (ME), and body condition score (BCS). Ewes fed the 37% RUP diet gained more (p<0.05) weight compared with ewes fed the 31% RUP diet (5.62 vs. 2.5kg). Ewes in EESM 60 had the highest levels of fat, protein, total solid, solid not fat, and immunoglobulin and the lowest in urea N content (P< 0.05) in colostrum during the first 24hrs after lambing. Conclusions: Protein source and RUP levels in ewes’ diets had no significant effect (P< 0.05) on lambs’ birth weight and ewes' blood biochemical parameters. Increasing the RUP content of diet during late gestation resulted in an increase in colostrum constituents and its IgG level but had no effect on ewes’ performance and their lambs’ outcome.

Keywords: colostrum, ewes, lambs output, pregnancy, undegradable protein

Procedia PDF Downloads 50

Authors: N. Mahmoudnia, M. Khormali

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This experiment was conducted to determine the apparent protein digestibility of poultry by- product meal (PBPM) from two industrial poultry slaughter houses on Ross 308 male broiler chickens in independed comparisons. The experiment consisted of seven dietary treatments and three replicates per treatment with three broiler chickens per replicate in a completely randomized design. Dietary treatments consisted of a control corn- soybean diet, and levels 3, 6 and 9% PBPM produced by slaughter house 1 and levels 3, 6 and 9% PBPM produced by slaughter house 2. Chromic oxide was added to the experimental diets as indigestible marker. The apparent protein digestibility of each diet were determined with two methods of sample collection of ileum and excreta in 21-28 d of age. The results this experiment showed that use of PBPM had no significantly effect on performance of broiler chicks during period of experiments. The apparent protein digestibility of PBPM groups was significantly higher than control group by excreta sampling procedure (P<0.05). Using of PBPM 2 significantly (P<0.05) decreased the apparent protein digestibility values based on ileum sampling procedure vs control ( 85.21 vs 90.14).Based results of this experiment,it is possible to use of PBPM 1 in broiler chicken.

Keywords: poultry by-product meal, apparent protein digestibility, independed comparison, broiler chicken

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

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The aerial surface of plants colonized by Human Enteric Pathogens ()has been implicated in outbreaks of enteric diseases in humans. Practice of organic farming primarily using animal dung as manure and sewage water for irrigation are the most significant source of enteric pathogens on the surface of leaves, fruits and vegetables. The present work aims to have an insight into the molecular mechanism of interaction of Human Enteric Pathogens or their metabolites with cell wall receptors in plants. Tomato plants grown under aseptic conditions at 12 hours L/D photoperiod, 25±1°C and 75% RH were inoculated individually with S. fonticola and K. pneumonia. The leaves from treated plants were sampled after 24 and 48 hours of incubation. The cell wall and cytoplasmic proteins were extracted and isocratically separated on 1D SDS-PAGE. The sampled leaves were also subjected to formaldehyde treatment prior to isolation of cytoplasmic proteins to study protein-protein interactions induced by Human Enteric Pathogens. Protein bands extracted from the gel were subjected to MALDI-TOF-TOF MS analysis. The foremost interaction of Human Enteric Pathogens on the plant surface was found to be cell wall bound receptors which possibly set ups a wave a critical protein-protein interaction in cytoplasm. The study revealed the expression and suppression of specific cytoplasmic and cell wall-bound proteins, some of them being important components of signaling pathways. The results also demonstrated HEP induced rearrangement of signaling pathways which possibly are crucial for adaptation of these pathogens to plant surface. At the end of the study, it can be concluded that controlling the over-expression or suppression of these specific proteins rearrange the signaling pathway thus reduces the outbreaks of food-borne illness.

Keywords: cytoplasmic protein, cell wall-bound protein, Human Enteric Pathogen (HEP), protein-protein interaction

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Authors: Arpit Kumar Shrivastava, Subrat Kumar, Rajani Kanta Mohapatra, Priyadarshi Soumyaranjan Sahu

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Computational approaches to predict structure, function and other biological characteristics of proteins are becoming more common in comparison to the traditional methods in drug discovery. Cryptosporidiosis is a major zoonotic diarrheal disease particularly in children, which is caused primarily by Cryptosporidium hominis and Cryptosporidium parvum. Currently, there are no vaccines for cryptosporidiosis and recommended drugs are not effective. With the availability of complete genome sequence of C. hominis, new targets have been recognized for the development of effective and better drugs and/or vaccines. We identified a unique hypothetical epitopic protein in C. hominis genome through BLASTP analysis. A 3D model of the hypothetical protein was generated using I-Tasser server through threading methodology. The quality of the model was validated through Ramachandran plot by PROCHECK server. The functional annotation of the hypothetical protein through DALI server revealed structural similarity with human Transportin 3. Phylogenetic analysis for this hypothetical protein also showed C. hominis hypothetical protein (CUV04613) was the closely related to human transportin 3 protein. The 3D protein model is further subjected to virtual screening study with inhibitors from the Zinc Database by using Dock Blaster software. Docking study reported N-(3-chlorobenzyl) ethane-1,2-diamine as the best inhibitor in terms of docking score. Docking analysis elucidated that Leu 525, Ile 526, Glu 528, Glu 529 are critical residues for ligand–receptor interactions. The molecular dynamic simulation was done to access the reliability of the binding pose of inhibitor and protein complex using GROMACS software at 10ns time point. Trajectories were analyzed at each 2.5 ns time interval, among which, H-bond with LEU-525 and GLY- 530 are significantly present in MD trajectories. Furthermore, antigenic determinants of the protein were determined with the help of DNA Star software. Our study findings showed a great potential in order to provide insights in the development of new drug(s) or vaccine(s) for control as well as prevention of cryptosporidiosis among humans and animals.

Keywords: cryptosporidium hominis, hypothetical protein, molecular docking, molecular dynamics simulation

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Authors: Mohammad Aladwan

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Alzheimer’s disease (AD) is a widespread dementing illness with a complex and poorly understood etiology. An important role in improving our understanding of the AD process is the modeling of disease-associated changes in tau protein phosphorylation, a protein known to mediate events essential to the onset and progression of AD. A main feature of AD is the abnormal phosphorylation of tau protein and the presence of neurofibrillary tangles. In order to evaluate the respective roles of the microtubule-binding region (MTBR) and alternatively spliced exons in the N-terminal projection domains in AD, we have constructed SHSY-5Y cell lines that stably overexpress four different species of tau protein (4R2N, 4R0N, N(E-2), N(E+2)). Since the toxicity and spreading of tau lesions in AD depends on the interactions of tau with other proteins, we have performed a proteomic analysis of exosome-fraction interactomes for cell lysates and media samples that were isolated from SHSY-5Y cell lines. Functional analysis of tau interactomes based on gene ontology (GO) terms was performed using the String 10.5 database program. The highest number of exosomes proteomes and tau associated proteins were found with 4R2N isoform (2771 and 159) in cell lysate and they have a high strength of connectivity (78%) between proteins, while N(E-2) isoform in the media proteomes has the highest number of proteins and tau associated protein (1829 and 205). Moreover, known AD markers were significantly enriched in secreted interactomes relative to lysate interactomes in the SHSY-5Y cells of tau isoforms lacking exons 2 and 3 in the N-terminal. The lack of exon 2 (E-2) from tau protein can be mediated by tau secretion and spreading to different cells. Enriched functions in the secreted E-2 interactome include signaling and developmental pathways that have been linked to a) tau misprocessing and lesion development and b) tau secretion and which, therefore, could play novel roles in AD pathogenesis.

Keywords: Alzheimer's disease, dementia, tau protein, neurodegenration disease

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Authors: Homayun Ghasemi, Mojtaba Yousefirad, Mozhgan Farzamisepehr

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Heavy metals are among soil pollutant resources that in case of accumulation in the soil and absorption by the plant, enter into the food chain and poison the plants or the people who consume those plants. This research was performed in order to examine the role of cadmium as a heavy metal in the activity of catalase and peroxidase as well as protein concentration in Trifolium resupinatum L. based on a randomized block design with three repetitions. The used treatments included consumption of Cd (NO3)2 at four levels, namely, 0, 100, 200, and 300 ppm. The plants under study were treated for 10 days. The results of the study showed that catalase activity decreased by the increase of cadmium. Moreover, peroxidase activity increased by an increase inthe consumption of cadmium. The analysis of protein level showed that plantlet protein decreased in high cadmium concentrations. The findings also demonstrated that cadmium concentration in roots was higher than in shoots.

Keywords: catalase, heavy metal, peroxidase, protein

Procedia PDF Downloads 247

Authors: Victor Prevost, Solene Landerneau, Michel Duhamel, Joel Sternheimer, Olivier Gallet, Pedro Ferrandiz, Marwa Mokni

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The search for homologous proteins is one of the ongoing challenges in biology and bioinformatics. Traditionally, a pair of proteins is thought to be homologous when they originate from the same ancestral protein. In such a case, their sequences share similarities, and advanced scientific research effort is spent to investigate this question. On this basis, we propose the Protein-Wave Alignment Tool (”P-WAT”) developed within the framework of the France Relance 2030 plan. Our work takes into consideration the mass-related wave aspect of protein biosynthesis, by associating specific frequencies to each amino acid according to its mass. Amino acids are then regrouped within their mass category. This way, our algorithm produces specific alignments in addition to those obtained with a common amino acid coding system. For this purpose, we develop the ”P-WAT” original algorithm, able to address large protein databases, with different attributes such as species, protein names, etc. that allow us to align user’s requests with a set of specific protein sequences. The primary intent of this algorithm is to achieve efficient alignments, in this specific conceptual frame, by minimizing execution costs and information loss. Our algorithm identifies sequence similarities by searching for matches of sub-sequences of different sizes, referred to as primers. Our algorithm relies on Boolean operations upon a dot plot matrix to identify primer amino acids common to both proteins which are likely to be part of a significant alignment of peptides. From those primers, dynamic programming-like traceback operations generate alignments and alignment scores based on an adjusted PAM250 matrix.

Keywords: protein, alignment, homologous, Genodic

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Authors: Roghaieh Parvizsedghy, Seyed Mojtaba Sadrameli

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Diminishing oil reserves, deteriorating health standards because of greenhouse gas emissions and associated environmental impacts have emerged biofuel production. Vegetable oils are proved to be valuable feedstock in these growing industries as they are renewable and potentially inexhaustible sources. Thermal Cracking of vegetable oils (triglycerides) leads to production of biofuels which are similar to fossil fuels in terms of composition but their combustion and physical properties have limits. Acrolein (very poisonous gas) and water production during cracking of triglycerides occurs because of presence of glycerin in their molecular structure. Transesterification of vegetable oil is a method to extract glycerol from triglycerides structure and produce methyl ester. In this study, castor methyl ester was used for thermal cracking in order to survey the efficiency of this method to produce bio-gasoline and bio-diesel. Thus, several experiments were designed by means of central composite method. Statistical studies showed that two reaction parameters, namely cracking temperature and feed flowrate, affect products yield significantly. At the optimized conditions (480 °C and 29 g/h) for maximum bio-gasoline production, 88.6% bio-oil was achieved which was distilled and separated as bio-gasoline (28%) and bio-diesel (48.2%). Bio-gasoline exposed a high octane number and combustion heat. Distillation curve and Reid vapor pressure of bio-gasoline fell in the criteria of standard gasoline (class AA) by ASTM D4814. Bio-diesel was compatible with standard diesel by ASTM D975. Water production was negligible and no evidence of acrolein production was distinguished. Therefore, thermal cracking of castor methyl ester could be used as a method to produce valuable biofuels.

Keywords: bio-diesel, bio-gasoline, castor methyl ester, thermal cracking, transesterification

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Authors: Friday A. Ogori, Ojotu M. Eke, Oneh J. Abu, Abraham T. Girgih

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B.aeqygtiaca del (Balanites aegyptiaca del) seed protein concentrate (APC) was hydrolyzed using different enzymes such as pepsin+pancreatin (PP), Alcalase (Alca), and Flavourzyme (Flav). The Alca has higher yield (100%) when compared to PP (83.23%) and Flav (62.90%). The hydrophobic amino acid and Sulphur containing amino acid (40.19%, 7.04%) in PP hydrolysate were higher compared to Alcalase (38.92%, 6.69%), Flavourenzyme (37.43%,6.35%), and APC (39.97%, 6.95%) samples. The PP has stronger DPPH, Hydroxyl radical quenching, Ferric reducing activity, and linoleic acid peroxidation activity, followed by the protein concentrate (APC) and Alcalase (Alca), while Flavourenzyme (Flav) derived hydrolysate was least in scavenging and inhibiting radical peroxidation properties. Flavourenzyme derived hydrolysate had stronger Ferric reducing antioxidant potential and metal chelating property. The result showed that the Alcalase hydrolysate has promising peptide yield, and PP hydrolysate had excellent amino acid residues and good in-vitro antioxidant potentials and could be a preferred ingredients in the nutraceutical and functional food emerging industries.

Keywords: balanites aegyptiaca del, protein concentrate, protein hydrolysates, enzymatic hydrolysis, antioxidants

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Authors: Vladimir Zobov, Nail Nazarov, Alexandra Vyshtakalyuk, Vyacheslav Semenov, Irina Galyametdinova, Vladimir Reznik

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The aim of this study was to investigate hepatoprotective properties of the Xymedon derivative L-ascorbate 1- (2-hydroxyethyl)-4,6-dimethyl-1,2-dihydropyrimidine-2-one (XD), which exhibits high efficiency as actoprotector. The study was carried out on 68 male albino rats weighing 250-400 g using preventive exposure to the test preparation. Effectiveness of XD win comparison with effectiveness of Xymedon (original substance) after administration of the compounds in identical doses. Maximum dose was 20 mg/kg. The animals orally received Xymedon or its derivative in doses of 10 and 20 mg/kg over 4 days. In 1-1.5 h after drug administration, CCl4 in vegetable oil (1:1) in a dose of 2 ml/kg. Controls received CCl4 but without hepatoprotectors. Intact control group consisted of rats, not receiving CCl4 or other compounds. The next day after the last administration of CCl4 and compounds under study animals were dehematized under ether anesthesia, blood and liver samples were taken for biochemical and histological analysis. Xymedon and XD administered according to the preventice scheme, exerted hepatoprotective effects: Xymedon — in the dose of 20 mg/kg, XD — in doses of 10 and 20 mg/kg. The drugs under study had different effects on liver condition, affected by induction with CCl4. Xymedon had a more pronounced effect both on the ALT level, which can be elevated not only due to destructive changes in hepatocytes, but also as a cholestasis manifestation, and on the serum total protein level, which reflects protein synthesis in liver. XD had a more pronounced effect on AST level, which is one of the markers of hepatocyte damage. Lower effective dose of XD — 10 mg/kg, compared to Xymedon effective according to, and its pronounced effect on AST, the hepatocyte cytolysis marker, is indicative of its higher preventive effectiveness, compared to Xymedon. This work was performed with the financial support of Russian Science Foundation (grant No: 14-50-00014).

Keywords: hepatoprotectors, pyrimidine derivatives, toxic liver damage, xymedon

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Authors: Thanusha D. Abeywickrama, Inoka C. Perera, Genji Kurisu

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Tuberculosis, considered being as the ninth leading cause of death worldwide, cause from a single infectious agent M. tuberculosis and the drug resistance nature of this bacterium is a continuing threat to the world. Therefore TB preventing treatment is expanding, where this study designed to analyze the regulatory mechanism of GntR transcriptional regulator gene Rv0792c, which lie between several genes codes for some hypothetical proteins, a monooxygenase and an oxidoreductase. The gene encoding Rv0792c was cloned into pET28a and expressed protein was purified to near homogeneity by Nickel affinity chromatography. It was previously reported that the protein binds within the intergenic region (BS region) between Rv0792c gene and monooxygenase (Rv0793). This resulted in binding of three protein molecules with the BS region suggesting tight control of monooxygenase as well as its own gene. Since monooxygenase plays a key role in metabolism, this gene may have a global regulatory role. The natural ligand for this regulator is still under investigation. In relation to the Rv0792 protein structure, a Circular Dichroism (CD) spectrum was carried out to determine its secondary structure elements. Percentage-wise, 17.4% Helix, 21.8% Antiparallel, 5.1% Parallel, 12.3% turn and 43.5% other were revealed from CD spectrum data under room temperature. Differential Scanning Calorimetry (DSC) was conducted to assess the thermal stability of Rv0792, which the melting temperature of protein is 57.2 ± 0.6 °C. The graph of heat capacity (Cp) versus temperature for the best fit was obtained for non-two-state model, which concludes the folding of Rv0792 protein occurs through stable intermediates. Peak area (∆HCal ) and Peak shape (∆HVant ) was calculated from the graph and ∆HCal / ∆HVant was close to 0.5, suggesting dimeric nature of the protein.

Keywords: CD spectrum, DSC analysis, GntR transcriptional regulator, protein structure

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Authors: Jaturong Matidtor, Krisna R. Torrissen, Saengtong Pongjareankit, Sudaporn Tongsiri, Jiraporn Rojtinnakorn

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Low survival rate has being particular problem in nursery of marble goby juvenile. The aim of this study was to investigate effect of garlic extract on protein digestive pancreatic enzymes, trypsin (T) and chymotrypsin (C). The marble goby were reared with commercial feed mixed garlic extract at concentration of 0 (control), 0.3, 0.5, 1.0, 3.0 and 5.0% (w/w) for 6 weeks. Analysis of the digestive enzymes at 2 and 6 weeks was performed. Growth parameters; weight gain (WG), specific growth rate (SGR) and feed efficiency (FE), were identified. For T, C and T/C at 2 weeks, values of T and T/C ratio of 0.3% (w/w) group showed significant difference (p < 0.05) with the highest values of 17685.64± 11981.77 U/mg protein and of 51.64 ± 27.46 U/mg protein, respectively. For C at 2 weeks, 0% (w/w) group showed the highest values of 16191.76± 2225.56 U/mg protein. Whereas value of T, C and T/C ratio at 6 weeks, there was no significant difference (p > 0.05). For growth performance, it significantly increased in all garlic extract fed groups (0.3-5.0%, w/w), both at 2 and 6 weeks. At 2 weeks, values of WG and SGR of 0.5% (w/w) group showed the highest values of 71.51 ± 1.60%, and 3.85 ± 0.07%, respectively. For FE, 0.3% (w/w) group showed the highest value of 60.21 ± 6.51%. At 6 weeks, it illustrated that all growth parameters of 5.0% (w/w) group were the highest values; WG = 35.06 ± 5.66%, SGR = 2.14 ± 0.30%, and FE = 5.86 ± 0.68%. We suggested that garlic extract could be available for protein digestive enzyme and growth enhancement in marble goby nursery with artificial feed. This result will be high benefit for commercial aquaculture of marble goby.

Keywords: marble goby, nursery, garlic extract, digestive enzyme, growth

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Authors: Rizky Kusuma Cahyani

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Hepatitis B is a liver inflammatory disease caused by hepatitis B virus (HBV). This infection can be prevented by vaccination which contains HBV surface protein (sHBsAg). However, vaccine supply is limited. Several attempts have been conducted to produce local sHBsAg. However, the purity degree and protein yield are still inadequate. Therefore optimization of HBsAg purification steps is required to obtain high yield with better purification fold. In this study, optimization of purification was done in 2 steps, precipitation using variation of NaCl concentration (0,3 M; 0,5 M; 0,7 M) and PEG (3%, 5%, 7%); ion exchange chromatography (IEC) using NaCl 300-500 mM elution buffer concentration.To determine HBsAg protein, bicinchoninic acid assay (BCA) and enzyme-linked immunosorbent assay (ELISA) was used in this study. Visualization of HBsAg protein was done by SDS-PAGE analysis. Based on quantitative analysis, optimal condition at precipitation step was given 0,3 M NaCl and PEG 3%, while in ion exchange chromatography step, the optimum condition when protein eluted with NaCl 500 mM. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis indicates that the presence of protein HBsAg with a molecular weight of 25 kDa (monomer) and 50 kDa (dimer). The optimum condition for purification of sHBsAg produced in Pichia pastoris gave a yield of 47% and purification fold 17x so that it would increase the production of hepatitis B vaccine to be more optimal.

Keywords: hepatitis B virus, HBsAg, hepatitis B surface antigen, Pichia pastoris, purification

Procedia PDF Downloads 151

Authors: S. Bilal, S. A. Bhat, I. Hussain, J. D. Parrah, S. P. Ahmad, M. R. Mir

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Background: The wound healing involves a highly coordinated cascade of cellular and immunological response over a period including coagulation, inflammation, granulation tissue formation, epithelialization, collagen synthesis and tissue remodeling. Wounds in aged heal more slowly than those in younger, mainly because of comorbidities that occur as one age. The present study is about the influence of age on wound healing. 1x1cm^2 (100 mm) wounds were created on the back of the animal. The animals were divided into two groups; one group had animals in the age group of 3-9 months while another group had animals in the age group of 15-21 months. Materials and Methods: 24 clinically healthy rabbits in the age group of 3-21 months were used as experimental animals and divided into two groups viz A and B. All experimental parameters, i.e., Excision wound model, Measurement of wound area, Protein extraction and estimation, Protein extraction and estimation and DNA extraction and estimation were done by standard methods. Results: The parameters studied were wound contraction, hydroxyproline, glucosamine, protein, and DNA. A significant increase (p<0.005) in the hydroxyproline, glucosamine, protein and DNA and a significant decrease in wound area (p<0.005) was observed in the age group of 3-9 months when compared to animals of an age group of 15-21 months. Wound contraction together with hydroxyproline, glucosamine, protein and DNA estimations suggest that advanced age results in retarded wound healing. Conclusion: The decrease wound contraction and accumulation of hydroxyproline, glucosamine, protein and DNA in group B animals may be associated with the reduction or delay in growth factors because of the advancing age.

Keywords: age, wound healing, excision wound, hydroxyproline, glucosamine

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Authors: Şevket Evci, Mehmet Akif Karsli

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The aim of this study is to determine nutrient contents, in situ ruminal degradation kinetics and protein fractions of screenings bean (B), chick pea (ChP), red lentil (RL) and green lentil (GL) that is used as residue in grain legume packing industry. For this purpose, four samples of each legumes species-a total of 16 samples, collected from different parts of our country were utilized. Feedstuffs used in the experiment were incubated for 0, 2 4, 8, 12, 24, and 48 hours in the rumen of 3 ruminally cannulated Akkaraman rams as duplicate. The nutrient contents, in situ ruminal dry matter (DM), organic matter (OM) and crude protein (CP) degradabilities and fractions, and escape protein contents were evaluated. The highest OM and CP contents were observed in RL (P<0.05). Chick pea had the highest ether extract (EE) content and EE values were 3.47, 6.72, 2.26, 8.66 % for RL, B, GL and ChP, respectively (P<0.05). Crude fiber (CF), ADF, and NDF contents were the highest in RL and the lowest in ChP. CF values were 24.03, 10.80, 4.09 and 3.57 % for RL, GL, B and ChP (P<0.05). Acid detergent insoluble nitrogen content of samples did not differ. Escape protein content was the highest in RL and the lowest in B (P<0.05). After 48 h incubation, the lowest OM and CP degradabilities were observed in RL. While the highest OM degradability was seen in ChP the highest CP degradability was observed in B (P<0.05). The lowest water soluble OM and CP contents were observed in RL whereas the highest potentially degradable OM and CP contents were seen in B and ChP (P<0.05). Both rate of OM and CP degradations (k-1) did not differ among samples (P>0.05). In conclusion, it was noted that feedstuffs (GL, ChP and B) used in the experiment except RL had a greater ruminal degradibilities of both OM and CP and moreover, had a higher escape protein contents, except B. It was thought that these feedstuffs can be substituted with some of common protein sources used in animal nutrition.

Keywords: in situ, nutrient contents, ruminant, subsieve

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Authors: Lavinia Ruta, Ileana Farcasanu

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The yeast surface display (YSD) technique enables the expression of proteins on yeast cell surfaces, facilitating the identification and isolation of proteins with targeted binding properties, such as nanobodies. Nanobodies, derived from camelid species, are single-domain antibody fragments renowned for their high affinity and specificity towards target proteins, making them valuable in research and potentially in therapeutics. Their advantages include a compact size (~15 kDa), robust stability, and the ability to target challenging epitopes. The project endeavors to establish and validate a platform for producing Green Fluorescent Protein (GFP) and mCherry nanobodies using the yeast surface display method. mCherry, a prevalent red fluorescent protein sourced from coral species, is commonly utilized as a genetic marker in biological studies due to its vibrant red fluorescence. The GFP-nanobody, a single variable domain of heavy-chain antibodies (VHH), exhibits specific binding to GFP, offering a potent means for isolating and engineering fluorescent protein fusions across various biological research domains. Both GFP and mCherry nanobodies find specific utility in cellular imaging and protein analysis applications.

Keywords: YSD, nanobodies, GFP, Saccharomyces cerevisiae

Procedia PDF Downloads 61

Authors: Vineeta Kaushik, Manisha Goel

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Chaperones are proteins that help other proteins fold correctly, and are found in all domains of life i.e., prokaryotes, eukaryotes and archaea. Various comparative genomic studies have suggested that the archaeal protein folding machinery appears to be highly similar to that found in eukaryotes. In case of protein folding; slow rotation of peptide prolyl-imide bond is often the rate limiting step. Formation of the prolyl-imide bond during the folding of a protein requires the assistance of other proteins, termed as peptide prolyl cis-trans isomerases (PPIases). Cyclophilins constitute the class of peptide prolyl isomerases with a wide range of biological function like protein folding, signaling and chaperoning. Most of the cyclophilins exhibit PPIase enzymatic activity and play active role in substrate protein folding which classifies them as a category of molecular chaperones. Till date, there is not very much data available in the literature on archaeal cyclophilins. We aim to compare the structural and biochemical features of the cyclophilin protein from within the three domains to elucidate the features affecting their stability and enzyme activity. In the present study, we carry out in-silico analysis of the cyclophilin proteins to predict their conserved residues, sites under positive selection and compare these proteins to their bacterial and eukaryotic counterparts to predict functional divergence. We also aim to clone and express these proteins in heterologous system and study their biophysical characteristics in detail using techniques like CD and fluorescence spectroscopy. Overall we aim to understand the features contributing to the folding, stability and dynamics of the archaeal cyclophilin proteins.

Keywords: biophysical characterization, x-ray crystallography, chaperone-like activity, cyclophilin, PPIase activity

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Authors: Meenakshi Verma, Mandeep Singh Bakshi, Kultar Singh

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Magnetic nanoparticles have received incredible importance in view of their diverse applications, which arise primarily due to their response to the external magnetic field. The magnetic behaviour of magnetic nanoparticles (NPs) helps them in numerous different ways. The most important amongst them is the ease with which they can be purified and also can be separated from the media in which they are present merely by applying an external magnetic field. This exceptional ease of separation of the magnetic NPs from an aqueous media enables them to use for extracting/removing metal pollutants from complex aqueous medium. Functionalized magnetic NPs can be subjected for the metallic impurities extraction if are favourably adsorbed on the NPs surfaces. We have successfully used the magnetic NPs as vehicles for gold and silver NPs removal from the complex fluids. The NPs loaded with gold and silver NPs pollutant fractions has been easily removed from the aqueous media by using external magnetic field. Similarly, we have used the magnetic NPs for extraction of protein from complex media and then constantly washed with pure water to eliminate the unwanted surface adsorbed components for quantitative estimation. The purified and protein loaded magnetic NPs are best analyzed with SDS Page to not only for characterization but also for separating the protein fractions. A collective review of the results indicates that we have synthesized surfactant coated iron oxide NPs and then functionalized these with selected materials. These surface active magnetic NPs work very well for the extraction of metallic NPs from the aqueous bulk and make the whole process environmentally sustainable. Also, magnetic NPs-Au/Ag/Pd hybrids have excellent protein extracting properties. They are much easier to use in order to extract the magnetic impurities as well as protein fractions under the effect of external magnetic field without any complex conventional purification methods.

Keywords: magnetic nanoparticles, protein, functionalized, extraction

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