Search results for: soluble protein

Authors: Muhammad Naeem, Amina Zubari, Abdus Salam, Syed Ali Ayub Bukhari, Naveed Ahmad Khan

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Seventy three wild Labeo bata of different body sizes, ranging from 8.20-16.00 cm total length and 7.4-86.19 g body weight, were studied for the analysis of body composition parameters (Water content, ash content, fat content, protein content) in relation to body size and condition factor. Mean percentage is found as for water 77.71 %, ash 3.42 %, fat 2.20 % and protein content 16.65 % in whole wet body weight. Highly significant positive correlations were observed between condition factor and body weight (r = 0.243). Protein contents, organic content and ash (% wet body weight) increase with increasing percent water contents for Labeo bata while these constituents (% dry body weight) and fat contents (% wet and dry body weight) have no influence on percent water. It was observed that variations in the body constituents have no association to body weight or length.

Keywords: Labeo bata, body size, body composition, condition factor

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Authors: Arfan Ali, Idrees Ahmad Nasir

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Potato (Solanum tuberosum) is an important cash crop and popular vegetable in Pakistan and throughout the world. Cold storage of potatoes accelerates the conversion of starch into reduced sugars (glucose and fructose). This process causes dry mass and bitter taste in the potatoes that are not acceptable to end consumers. In the current study, the phosphofructokinase B gene was cloned into the pET-30 vector for protein expression and the pCambia-1301 vector for plant expression. Amplification of a 930bp product from an E. coli strain determined the successful isolation of the phosphofructokinase B gene. Restriction digestion using NcoI and BglII along with the amplification of the 930bp product using gene specific primers confirmed the successful cloning of the PfkB gene in both vectors. The protein was expressed as a His-PfkB fusion protein. Western blot analysis confirmed the presence of the 35 Kda PfkB protein when hybridized with anti-His antibodies. The construct Fani-01 was evaluated transiently using a histochemical gus assay. The appearance of blue color in the agroinfiltrated area of potato leaves confirmed the successful expression of construct Fani-01. Further, the area displaying gus expression was evaluated for PfkB expression using ELISA. Moreover, PfkB gene expression evaluated through transient expression determined successful gene expression and highlighted its potential utilization for stable expression in potato to reduce sweetening due to long-term storage.

Keywords: potato, Solanum tuberosum, transformation, PfkB, anti-sweetening

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Authors: E. Obreque-Slier, F. Orellana-Rodríguez, R. López-Solís

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Phenolic compounds are secondary metabolites present in some foods, such as wine. Polyphenols comprise two main groups: flavonoids (anthocyanins, flavanols, and flavonols) and non-flavonoids (stilbenes and phenolic acids). Phenolic acids are low molecular weight non flavonoid compounds that are usually grouped into benzoic (gallic, vanillinic and protocatechuic acids) and cinnamic acids (ferulic, p-coumaric and caffeic acids). Likewise, tannic acid is an important polyphenol constituted mainly by gallic acid. Phenolic compounds are responsible for important properties in foods and drinks, such as color, aroma, bitterness, and astringency. Astringency is a drying, roughing, and sometimes puckering sensation that is experienced on the various oral surfaces during or immediately after tasting foods. Astringency perception has been associated with interactions between flavanols present in some foods and salivary proteins. Despite the quantitative relevance of phenolic acids in food and beverages, there is no information about its effect on salivary proteins and consequently on the sensation of astringency. The objective of this study was assessed the interaction of several phenolic acids (gallic, vanillinic, protocatechuic, ferulic, p-coumaric and caffeic acids) with saliva. Tannic acid was used as control. Thus, solutions of each phenolic acids (5 mg/mL) were mixed with human saliva (1:1 v/v). After incubation for 5 min at room temperature, 15-μL aliquots of the mixtures were dotted on a cellulose membrane and allowed to diffuse. The dry membrane was fixed in 50 g/L trichloroacetic acid, rinsed in 800 mL/L ethanol and stained for protein with Coomassie blue for 20 min, destained with several rinses of 73 g/L acetic acid and dried under a heat lamp. Both diffusion area and stain intensity of the protein spots were semiqualitative estimates for protein-tannin interaction (diffusion test). The rest of the whole saliva-phenol solution mixtures of the diffusion assay were centrifuged and fifteen-μL aliquots of each supernatant were dotted on a cellulose membrane, allowed to diffuse and processed for protein staining, as indicated above. In this latter assay, reduced protein staining was taken as indicative of protein precipitation (precipitation test). The diffusion of the salivary protein was restricted by the presence of each phenolic acids (anti-diffusive effect), while tannic acid did not alter diffusion of the salivary protein. By contrast, phenolic acids did not provoke precipitation of the salivary protein, while tannic acid produced precipitation of salivary proteins. In addition, binary mixtures (mixtures of two components) of various phenolic acids with gallic acid provoked a restriction of saliva. Similar effect was observed by the corresponding individual phenolic acids. Contrary, binary mixtures of phenolic acid with tannic acid, as well tannic acid alone, did not affect the diffusion of the saliva but they provoked an evident precipitation. In summary, phenolic acids showed a relevant interaction with the salivary proteins, thus suggesting that these wine compounds can also contribute to the sensation of astringency.

Keywords: astringency, polyphenols, tannins, tannin-protein interaction

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Authors: Priya Arora, Ashutosh Mishra

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Gene identification, towards the pursuit of mutated genes, leading to Parkinson’s disease, puts forward a challenge towards proactive cure of the disorder itself. Computational analysis is an effective technique for exploring genes in the form of protein sequences, as the theoretical and manual analysis is infeasible. The limitations and effectiveness of a particular computational method are entirely dependent on the previous data that is available for disease identification. The article presents a sequence-based classification method for the identification of genes responsible for Parkinson’s disease. During the initiation phase, the physicochemical properties of amino acids transform protein sequences into a feature vector. The second phase of the method employs Jaccard distances to select negative genes from the candidate population. The third phase involves artificial neural networks for making final predictions. The proposed approach is compared with the state of art methods on the basis of F-measure. The results confirm and estimate the efficiency of the method.

Keywords: disease gene identification, Parkinson’s disease, physicochemical properties of amino acid, protein sequences

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Authors: Harleen Kaur, Richa

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Plants produce a large variety of secondary metabolites. Phenolics are the compounds that contain hydroxyl functional group on an aromatic ring. These are chemically heterogeneous compounds. Some are soluble only in organic solvents, some are water soluble and others are large insoluble polymers. Flavonoids are one of the largest classes of plant phenolics. The carbon skeleton of a flavonoid contains 15 carbons arranged in two aromatic rings connected by a three carbon ridge. Both phenolics and flavonoids are good natural antioxidants. Four Indian aromatic plants were selected for the study i.e, Achillea species, Jasminum primulinum, Leucas cephalotes and Leonotis nepetaefolia. All the plant species were collected from Chail region of Himachal Pradesh, India. The identifying features and anatomical studies were done of the part containing the essential oils. Phenolic cotent was estimated by Folin Ciocalteu’s method and flavonoids content by aluminium chloride method. Antioxidant property was checked by using DPPH method. Maximum antioxidant potential was found in Achillea species, followed by Leonotis nepetaefolia, Jaminum primulinum and Leucas cephalotes. Phenolics and flavonoids are important compounds that serve as defences against herbivores and pathogens. Others function in attracting pollinators and absorbing harmful radiations.

Keywords: antioxidants, DPPH, flavonoids, phenolics

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Authors: Kinnsley Travis, Camerron M. Crowder

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Mapk8ip3 (Mitogen-Activated Protein Kinase 8 Interacting Protein 3) is a gene that codes for the JIP3 protein, which is a part of the JIP scaffolding protein family. This protein is involved in axonal vesicle transport, elongation and regeneration. Variants in the Mapk8ip3 gene are associated with a rare-genetic condition that results in a neurodevelopmental disorder that can cause a range of phenotypes including global developmental delay and intellectual disability. Currently, there are 18 known individuals diagnosed to have sequenced confirmed Mapk8ip3 genetic disorders. This project focuses on examining the impact of a subset of missense patient variants on the Jip3 protein function by overexpressing the mRNA of these variants in a zebrafish knockout model for Jip3. Plasmids containing cDNA with individual missense variants were reverse transcribed, purified, and injected into single-cell zebrafish embryos (Wild Type, Jip3 -/+, and Jip3 -/-). At 6-days post mRNA microinjection, morphological, behavioral, and microscopic phenotypes were examined in zebrafish larvae. Morphologically, we compared the size and shape of the zebrafish during their development over a 5-day period. Total locomotive activity was assessed using the Microtracker assay and patterns of movement over time were examined using the DanioVision assay. Lastly, we used confocal microscopy to examine sensory axons for swelling and shortened length, which are phenotypes observed in the loss-of-function knockout Jip3 zebrafish model. Using these assays during embryonic development, we determined the impact of various missense variants on Jip3 protein function, compared to knockout and wild-type zebrafish embryo models. Variants in the gene Mapk8ip3 cause rare-neurodevelopmental disorders due to an essential role in axonal vesicle transport, elongation and regeneration. A subset of missense variants was examined by overexpressing the mRNA of these variants in a Jip3 knock-out zebrafish. Morphological, behavioral, and microscopic phenotypes were examined in zebrafish larvae. Using these assays, the spectrum of disorders can be phenotypically determined and the impact of variant location can be compared to knockout and wild-type zebrafish embryo models.

Keywords: rare disease, neurodevelopmental disorders, mrna overexpression, zebrafish research

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Authors: Sema Şenoğlu, Sevgi Karakuş

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Hydrazone scaffold is important to design new drug groups and is found to possess numerous uses in pharmaceutical chemistry. Besides, hydrazone derivatives are also known for biological activities such as anticancer, antimicrobial, antiviral, and antifungal. Hydrazone derivatives are promising anticancer agents because they inhibit cancer proliferation and induce apoptosis. Human carbonic anhydrase IX has a high potential to be an antiproliferative drug target, and targeting this protein is also important for obtaining potential anticancer inhibitors. The protein construct was retrieved as a PDB file from the RCSB protein database. This binding interaction of proteins and ligands was performed using Discovery Studio Visualizer. In vitro inhibitory activity of hydrazone derivatives was tested against enzyme carbonic anhydrase IX on the PyRx programme. Most of these molecules showed remarkable human carbonic anhydrase IX inhibitory activity compared to the acetazolamide. As a result, these compounds appear to be a potential target in drug design against human carbonic anhydrase IX.

Keywords: cancer, carbonic anhydrase IX enzyme, docking, hydrazone

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Authors: Aljabryn Dalal Hamad

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The present explanatory study concerns with the relation between Diabetes Mellitus and Food Balance in the Kingdom of Saudi Arabia during 2005-2010, using published data. Results illustrated that Saudi citizen daily protein consumption (DPC) during 2005-2007 (g/capita/day) is higher than the average global consumption level of protein with 15.27%, daily fat consumption (DFC) with 24.56% and daily energy consumption (DEC) with 16.93% and increases than recommended level by International Nutrition Organizations (INO) with 56% for protein, 60.49% for fat and 27.37% for energy. On the other hand, DPC per capita in Saudi Arabia decreased during the period 2008-2010 from 88.3 to 82.36 gram/ day. Moreover, DFC per capita in Saudi Arabia decreased during the period 2008-2010 from 3247.90 to 3176.43 Cal/capita/ day, and daily energy consumption (DEC) of Saudi citizen increases than world consumption with 16.93%, while increases with 27.37% than INO. Despite this, DPC, DFC and DEC per capita in Saudi Arabia still higher than world mean. On the other side, results illustrated that the number of diabetic patients in Saudi Arabia during the same period (2005-2010). The curve of diabetic patient’s number in Saudi Arabia during 2005-2010 is regular ascending with increasing level ranged between 7.10% in 2005 and 12.44% in 2010. It is essential to devise Saudi National programs to educate the public about the relation of food balances and diabetes so it could be avoided, and provide citizens with healthy dietary balances tables.

Keywords: Diabetes mellitus, food balance, energy, fat, protein, Saudi Arabia

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Authors: Jiang Xu, Daoping Wang, Yinghong Pan

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The jointing-booting stage is a critical period of both vegetative growth and reproductive growth in rice. Therefore, the proteomic analysis of the mutant Osbri1, whose corresponding gene OsBRI1 encodes the putative BRs receptor OsBRI1, at jointing-booting stage is very important for understanding the effects of BRs on vegetative and reproductive growth. In this study, the proteomes of leaves from an allelic mutant of the DWARF 61 (D61, OsBRI1) gene, Fn189 (dwarf54, d54) and its wild-type variety T65 (Taichung 65) at jointing-booting stage were analysed by using a Q Exactive plus orbitrap mass spectrometer, and more than 3,100 proteins were identified in each sample. Ontology analysis showed that these proteins distribute in various space of the cells, such as the chloroplast, mitochondrion, and nucleus, they functioned as structural components and/or catalytic enzymes and involved in many physiological processes. Moreover, quantitative analysis displayed that 266 proteins were differentially expressed in two samples, among them, 77 proteins decreased and 189 increased more than two times in Fn189 compared with T65, the proteins whose content decreased in Fn189 including b5-like Heme/Steroid binding domain containing protein, putative retrotransposon protein, putative glutaminyl-tRNA synthetase, and higher content proteins such as mTERF, putative Oligopeptidase homologue, zinc knuckle protein, and so on. A former study founded that the transcription level of a mTERF was up-regulated in the leaves of maize seedling after EBR treatment. In our experiments, it was interesting that one mTERF protein increased, but another mTERF decreased in leaves of Fn189 at jointing-booting stage, which suggested that BRs may have differential regulation mechanisms on the expression of various mTERF proteins. The relationship between other differential proteins with BRs is still unclear, and the effects of BRs on rice protein contents and its regulation mechanisms still need further research.

Keywords: bri1 mutant, jointing-booting stage, proteomic analysis, rice

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Authors: Afiqah Shafify Amran, Siti Aisyah Shamsudin, Nurul Yuziana Mohd Yusof

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Cadmium Sulphide (CdS) from group II-IV quantum dots with good optical properties was successfully synthesized by using the simple colloidal method. Capping them with ligand Polyethylinamine (PEI) alters the surface defect of CdS while, thioglycolic acid (TGA) was added to the reaction as a stabilizer. Due to their cytotoxicity, we decided to conjugate them with the protein cage nanoparticles. In this research, we used capsid of Cowpea Chlorotic Mottle Virus (CCMV) to package the CdS because they have the potential to serve in drug delivery, cell targeting and imaging. Adding Sodium Hydroxide (NaOH) changes the pH of the systems hence the isoelectric charge is adjusted. We have characterized and studied the morphology and the optical properties of CdS and CdS-CCMV by transmitted electron microscopic (TEM), UV-Vis spectroscopy, photoluminescence spectroscopy, UV lamp and Fourier transform infrared spectroscopy (FTIR), respectively. The results obtained suggest that the protein cage nanoparticles do not affect the optical properties of CdS.

Keywords: cadmium sulphide, cowpea chlorotic mottle virus, protein cage nanoparticles, quantum dots

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Authors: Abdelsabour G. A. Khaled, Ahmed W. A. Abdalla And Sabry Y. M. Mahmoud

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Banana plants showing typical mosaic and yellow stripes on leaves as symptoms were collected from Assiut Governorate in Egypt. The causal agent was identified as Cucumber mosaic virus (CMV) on the basis of symptoms, transmission, serology, transmission electron microscopy and reverse transcription polymerase chain reaction (RT-PCR). Coat protein (CP) gene was amplified using gene specific primers for coat protein (CP), followed by cloning into desired cloning vector for sequencing. In this study the CMV was transmitted into propagation host either by aphid or mechanically. The transmission was confirmed through Direct Antigen Coating Enzyme Linked Immuno Sorbent Assay (DAC-ELISA). Analysis of the 120 deduced amino acid sequence of the coat protein gene revealed that the EG-A strain of CMV shared from 97.50 to 98.33% with those strains belonging to subgroup IA. The cluster analysis grouped the Egyptian isolate with strains Fny and Ri8 belonging sub-group IA. It appears that there occurs a high incidence of CMV infecting banana belonging to IA subgroup in most parts of Egypt.

Keywords: banana, CMV, transmission, CP gene, RT-PCR

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Authors: Saha Saradindu, Das Payel, Somdeb BoseDasgupta

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Tuberculosis caused by Mycobacteria tuberculosis is a dreadful disease and more so with the advent of extreme and total drug-resistant species. Mycobacterial pathogenesis is an ever-changing paradigm from phagosome maturation block to phagosomal escape into macrophage cytosol and finally acid tolerance and survival inside the lysosome. Mycobacteria are adept at subverting the host immune response by highjacking host cell signaling and secreting virulence factors. One such virulence factor is a ser/thr kinase; Protein kinase G (PknG), which is known to prevent phagosome maturation. The host substrates of PknG, allowing successful pathogenesis still remain an enigma. Hence we carried out a comparative phosphoproteomic screen and identified a number of substrates phosphorylated by PknG. We characterized some of these substrates in vivo and in vitro and observed that PknG mediated phosphorylation of these substrates leads to reduced TNFa production as well as decreased response to TNFa induced macrophage necroptosis, thus enabling mycobacterial survival and proliferation.

Keywords: mycobacteria, Protein kinase G, phosphoproteomics, necroptosis

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Authors: S. Barbaqadze, M. Goderdzishvili, E. Mosidze, L. Lomtadze, V. Mshvildadze, L. Bakuridze, D. Berashvili, A. Bakuridze

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This paper focuses on the formulation of phytobiotic designated for further implantation in poultry farming. Composition was meant to be water-soluble powder containing antibacterial essential oils. The development process involved Thyme, Monarda and Clary sage essential oils. The antimicrobial activity of essential oils composite was meant to be tested against gram-negative and gram-positive bacterial strains. The results are processed using the statistical program Sigma STAT. To make essential oils composition water soluble surfactants were added to them. At the first stage of the study, nine options for the optimal composition of essential oils and surfactants were developed. The effect of the amount of surfactants on the essential oils composition solubility in water has been investigated. On the basis of biopharmaceutical studies, the formulation of phytobiotic has been determined: Thyme, monarda and clary sage essential oils 2:1:1 - 100 parts; Licorice extract 5.25 parts and inhalation lactose 300 parts. A technology for the preparation of phytobiotic has been developed and a technological scheme for the preparation of phytobiotic has been made up. The research was performed within the framework of the grant project CARYS-19-363 funded be the Shota Rustaveli National Science Foundation of Georgia.

Keywords: clary, essential oils, monarda, phytobiotics, poultry, thyme

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Authors: Nonlapun Meenil, Pisitpong Intarapong, Thitima Wongsheree, Pranchalee Samanpiboon

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In Thailand, once pineapples are harvested, they must be classified into two classes based on their sweetness: sweet and unsweet. This paper has studied and developed the assessment of internal quality of pineapples using a low-cost compact spectroscopy sensor according to the Spectroscopy approach and Neural Network (NN). During the experiments, Batavia pineapples were utilized, generating 100 samples. The extracted pineapple juice of each sample was used to determine the Soluble Solid Content (SSC) labeling into sweet and unsweet classes. In terms of experimental equipment, the sensor cover was specifically designed to install the sensor and light source to read the reflectance at a five mm depth from pineapple flesh. By using a spectroscopy sensor, data on visible and near-infrared reflectance (Vis-NIR) were collected. The NN was used to classify the pineapple classes. Before the classification step, the preprocessing methods, which are Class balancing, Data shuffling, and Standardization were applied. The 510 nm and 900 nm reflectance values of the middle parts of pineapples were used as features of the NN. With the Sequential model and Relu activation function, 100% accuracy of the training set and 76.67% accuracy of the test set were achieved. According to the abovementioned information, using a low-cost compact spectroscopy sensor has achieved favorable results in classifying the sweetness of the two classes of pineapples.

Keywords: neural network, pineapple, soluble solid content, spectroscopy

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Authors: Khushi Kashyap, Pratibha Singh

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In the recent years increasing interest in vegetarian diets has been observed, a major problem in this type of diet is to provide the appropriate amount of protein .Value addition of food is current most talked topic because of increasing nutritional awareness among consumers today. An investigation was conducted to develop protein rich multi-millet hemp seed khakhra. The seeds of cannabis sativa L. have been a significant source of food for thousand of year. In recent years, hemp has not been thoroughly explored for its nutritional potential due to the mistaken belief regarding the cannabis plants. Methodology- two variations was prepared referencing standard recipe. Variation 1 was prepared using 25g ragi, 25g bajra,40g whole wheat flour with 10g hemp seed powder, variation 2(RF-25g,BF25g,WWF-35g,HS-15g). The product was subjected to sensory evolution by semi trained panel members using 9 point hedonic on 50 panelists. Result- result of the sensory evaluation revealed that the product incorporated with 15g of hemp seed were similar to control I texture, taste and overall quality and was more acceptable by the panelist and was selected as final product seed. On estimation of the nutrient content 30g of khakhra provides 107kcal of energy,12g protein,75g carbohydrate, and 9.6g of fats with shelf life of 3 months. Conclusion- khakhras can be eaten as a snack at any time of the day. hemp seed powder incorporated in it enhances its nutritive value and makes it more nutritious. It is suitable for consumption of all the age group.

Keywords: cannabis sativa, hemp, protein, seed

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Authors: Hung-Chieh Chen, Kuo-Hui Wang, Tsu-Lin Yeh

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Proteomics represent the performance of genomic complement proteins and the protein level on functional genomics. This study adopted proteomic strategies for comparing serum proteins among three groups: elite sprinter (sprint runner group, SR), long-distance runners (long-distance runner group, LDR), and the untrained control group (control group, CON). Purposes: This study aims to identify elite sprinters and long-distance runners’ serum protein and to provide a comparison of their serum proteome’ composition. Methods: Serum protein fractionations that separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and analyzed by a quantitative nano-LC-MS/MS-based proteomic profiling. The one-way analysis of variance (ANOVA) and Scheffe post hoc comparison (α= 0.05) was used to determine whether there is any significant difference in each protein level among the three groups. Results: (1) After analyzing the 307 identified proteins, there were 26 unique proteins in the SR group, and 18 unique proteins in the LDR group. (2) For the LDR group, 7 coagulation function-associated proteins’ expression levels were investigated: vitronectin, serum paraoxonase/arylesterase 1, fibulin-1, complement C3, vitamin K-dependent protein, inter-alpha-trypsin inhibitor heavy chain H3 and von Willebrand factor, and the findings show the seven coagulation function-associated proteins were significantly lower than the group of SR. (3) Comparing to the group of SR, this study found that the LDR group’s expression levels of the 2 antioxidant proteins (afamin and glutathione peroxidase 3) were also significantly lower. (4) The LDR group’s expression levels of seven immune function-related proteins (Ig gamma-3 chain C region, Ig lambda-like polypeptide 5, clusterin, complement C1s subcomponent, complement factor B, complement C4-A, complement C1q subcomponent subunit A) were also significantly lower than the group of SR. Conclusion: This study identified the potential serum protein markers for elite sprinters and long-distance runners. The changes in the regulation of coagulation, antioxidant, or immune function-specific proteins may also provide further clinical applications for these two different track athletes.

Keywords: biomarkers, coagulation, immune response, oxidative stress

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Authors: P. Lestinsky, D. Jecha, V. Brummer, P. Stehlik

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Scrubbing by a liquid spraying is one of the most effective processes used for removal of fine particles and soluble gas pollutants (such as SO2, HCl, HF) from the flue gas. There are many configurations of scrubbers designed to provide contact between the liquid and gas stream for effectively capturing particles or soluble gas pollutants, such as spray plates, packed bed towers, jet scrubbers, cyclones, vortex and venturi scrubbers. The primary function of venturi scrubber is the capture of fine particles as well as HCl, HF or SO2 removal with effect of the flue gas temperature decrease before input to the absorption column. In this paper, sulfur dioxide (SO2) from flue gas was captured using new design replacing venturi scrubber (1st degree of wet scrubbing). The flue gas was prepared by the combustion of the carbon disulfide solution in toluene (1:1 vol.) in the flame in the reactor. Such prepared flue gas with temperature around 150 °C was processed in designed laboratory O-element scrubber. Water was used as absorbent liquid. The efficiency of SO2 removal, pressure drop and temperature drop were measured on our experimental device. The dependence of these variables on liquid-gas ratio was observed. The average temperature drop was in the range from 150 °C to 40 °C. The pressure drop was increased with increasing of a liquid-gas ratio, but not as much as for the common venturi scrubber designs. The efficiency of SO2 removal was up to 70 %. The pressure drop of our new designed wet scrubber is similar to commonly used venturi scrubbers; nevertheless the influence of amount of the liquid on pressure drop is not so significant.

Keywords: desulphurization, absorption, flue gas, modeling

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Authors: Olaide Ruth Aderibigbe, Oluwatoyin Oluwole

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Pigeon pea is one of the minor leguminous plants; though underutilised, it is used traditionally by farmers to alleviate hunger and malnutrition. Pigeon pea is cultivated in Nigeria by subsistence farmers. It is rich in protein and minerals, however, its utilisation as food is only common among the poor and rural populace who cannot afford expensive sources of protein. One of the factors contributing to its limited use is the high antinutrient content which makes it indigestible, especially when eaten by children. The development of value-added products that can reduce the antinutrient content and make the nutrients more bioavailable will increase the utilisation of the crop and contribute to reduction of malnutrition. This research, therefore, determined the effects of different roasting temperatures (130 0C, 140 0C, and 150 0C) on the proximate, mineral and antinutrient component of a pigeon pea snack. The brown variety of pigeon pea seeds were purchased from a local market- Otto in Lagos, Nigeria. The seeds were cleaned, washed, and soaked in 50 ml of water containing sugar and salt (4:1) for 15 minutes, and thereafter the seeds were roasted at 130 0C, 140 0C, and 150 0C in an electric oven for 10 minutes. Proximate, minerals, phytate, tannin and alkaloid content analyses were carried out in triplicates following standard procedures. The results of the three replicates were polled and expressed as mean±standard deviation; a one-way analysis of variance (ANOVA) and the Least Significance Difference (LSD) were carried out. The roasting temperatures significantly (P<0.05) affected the protein, ash, fibre and carbohydrate content of the snack. Ready-to-eat snack prepared by roasting at 150 0C significantly had the highest protein (23.42±0.47%) compared the ones roasted at 130 0C and 140 0C (18.38±1.25% and 20.63±0.45%, respectively). The same trend was observed for the ash content (3.91±0.11 for 150 0C, 2.36±0.15 for 140 0C and 2.26±0.25 for 130 0C), while the fibre and carbohydrate contents were highest at roasting temperature of 130 0C. Iron, zinc, and calcium were not significantly (P<0.5) affected by the different roasting temperatures. Antinutrients decreased with increasing temperature. Phytate levels recorded were 0.02±0.00, 0.06±0.00, and 0.07±0.00 mg/g; tannin levels were 0.50±0.00, 0.57±0.00, and 0.68±0.00 mg/g, while alkaloids levels were 0.51±0.01, 0.78±0.01, and 0.82±0.01 mg/g for 150 0C, 140 0C, and 130 0C, respectively. These results show that roasting at high temperature (150 0C) can be utilised as a processing technique for increasing protein and decreasing antinutrient content of pigeon pea.

Keywords: antinutrients, pigeon pea, protein, roasting, underutilised species

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Authors: Yue He, Youn Young Shim, Ji Hye Kim, Jae Youl Cho, Martin J. T. Reaney

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Recently, pulse cooking water (a.k.a. Aquafaba) has been used as an important and cost-effective alternative to eggs in gluten-free, vegan cooking and baking applications. The aquafaba (AQ) is primarily due to its excellent ability to stabilize foams and emulsions in foods. However, the functional ingredients of this excellent AQ are usually discarded with the compound release. This study developed a high-functional food material, AQ, using functional soybean AQ that has not been studied in Korea. A zero-waste and cost-effective hybrid process were used to produce oil emulsifiers from Korean soybeans. The treatment technique was implemented using a small number of efficient steps. Aquafaba from Backtae had the best emulsion properties (92%) and has the potential to produce more stable food oil emulsions. Therefore, this study is expected to be utilized in the development of the first gluten-free, vegan product for vegetarians and consumers with animal protein allergies, utilizing wastewater from cooked soybeans as a source of plant protein that can replace animal protein.

Keywords: aquafaba, soybean, chickpea, emulsifiers, egg replacer, egg-free products

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Authors: Ningombam Linthoingambi Devi, Amrendra Kumar

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The long term sampling at one of the most populated city in Indo-Gangetic region shows higher mass concentration of atmospheric aerosol (PM₂.₅) during spring season (144.70µg/m³), summer season (91.96 µg/m³), the autumn season (266.48µg/m³) and winter season (367.09 µg/m³) respectively. The concentration of PM₂.₅ in Patna across the year shows much higher than the limit fixed by the national ambient air quality level fixed by central pollution control board India (CPCB, India) and World Health Organization (WHO). Different water-soluble cation (Na⁺, K⁺, Ca²⁺, NH₄⁺ , and Mg²⁺) and anion (Cl⁻, NO₃⁻ , and SO₄²⁻) species were detected in PM₂.₅. Results show the significantly higher loaded of water-soluble ions during winter and spring seasons. The acidity of the atmosphere was revealed and calculated using selected major cations (K⁺, Ca²⁺ , and NH₄⁺) and anions (SO₄²⁻, and NO₃⁻). A regression correlation was analyzed to check the significant linkage between the acidity and alkalinity ions. During the winter season (r² = 0.79) and spring season (r² = 0.64) shows good significant correlation between the cations and anions. The ratio of NO₃⁻/SO₄²⁻ indicates the sources of secondary pollutants were mainly influenced by industrial and vehicular emission however SO₄²⁻ mostly emitted from industries during the winter season.

Keywords: aerosols, inorganic species, source apportionment, Indo-Gangetic region

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Authors: A. Alyami, S. Christmas, K. Neeltje, G. Pollakis, B. Paxton, Z. Al-Bayati

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The Human leukocyte antigen-G (HLA-G) molecule plays an important role in immunomodulation. To date, 16 untranslated regions (UTR) HLA-G haplotypes have been previously defined by sequenced SNPs in the coding region. From these, UTR-1, UTR-2, UTR-3, UTR-4, UTR-5, UTR-6 and UTR-7 are the most frequent 3’UTR haplotypes at the global level. UTR-1 is associated with higher levels of soluble HLA-G and HLA-G expression, whereas UTR-5 and UTR-7 are linked with low levels of soluble HLA-G and HLA-G expression. Human immunodeficiency virus type 1 (HIV-1) infection results in the progressive loss of immune function in infected individuals. The virus escape mechanism typically includes T lymphocytes and NK cell recognition and lyses by classical HLA-A and B down-regulation, which has been associated with non-classical HLA-G molecule up-regulation, respectively. We evaluated the haplotypes of the HLA-G 3′ untranslated region frequencies observed in three HIV-1 groups from the Netherlands and their susceptibility to develop infection. The three groups are made up of mainly men who have sex with men (MSM), injection drug users (IDU) and a high-risk-seronegative (HRSN) group. DNA samples were amplified with published primers prior sequencing. According to our results, the low expresser frequencies show higher in HRSN compared to other groups. This is indicating that 3’UTR polymorphisms may be identified as potential prognostic biomarkers to determine susceptibility to HIV.

Keywords: Human leukocyte antigen-G (HLA-G) , men who have sex with men (MSM), injection drug users (IDU), high-risk-seronegative (HRSN) group, high-untranslated region (UTR)

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Authors: Jong Il Rhee

Abstract:

The polypeptide representing the mature part of human BMP-7 was cloned and efficiently expressed in Escherichia coli and Bacillus subtilis, which had a clear band for hBMP-7, a homodimeric protein with an apparent molecular weight of 15.4 kDa. Recombinant E.coli produced 111 pg hBMP-7/mg of protein hBMP-7 through IPTG induction. Recombinant B. subtilis also produced 350 pg hBMP-7/ml of culture medium. The hBMP-7 was purified in 2 steps using an FPLC system with an ion exchange column and a gel filtration column. The hBMP-7 produced in this work also stimulated the alkaline phosphatase (ALP) activity in a dose-dependent manner, i.e. 2.5- and 8.9-fold at 100 and 300 ng hBMP-7/ml, respectively, and showed intact biological activity.

Keywords: B. subtilis, E. coli, fermentation, hBMP-7

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Authors: Priyanka Jain, Ashish K. Sharma, Esha Shukla, K. J. Mukherjee

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We propose a paradigm shift in our approach to design improved platforms for recombinant protein production, by addressing system level issues rather than the individual steps associated with recombinant protein synthesis like transcription, translation, etc. We demonstrate that by controlling and modulating the cellular stress response (CSR), which is responsible for feedback control of protein synthesis, we can generate hyper-producing strains. We did transcriptomic profiling of post-induction cultures, expressing different types of protein, to analyze the nature of this cellular stress response. We found significant down-regulation of substrate utilization, translation, and energy metabolism genes due to generation CSR inside the host cell. However, transcription profiling has also shown that many genes are up-regulated post induction and their role in modulating the CSR is unclear. We hypothesized that these up-regulated genes trigger signaling pathways, generating the CSR and concomitantly reduce the recombinant protein yield. To test this hypothesis, we knocked out the up-regulated genes, which did not have any downstream regulatees, and analyzed their impact on cellular health and recombinant protein expression. Two model proteins i.e., GFP and L-Asparaginase were chosen for this analysis. We observed a significant improvement in expression levels, with some knock-outs showing more than 7-fold higher expression compared to control. The 10 best single knock-outs were chosen to make 45 combinations of all possible double knock-outs. A further increase in expression was observed in some of these double knock- outs with GFP levels being highest in a double knock-out ΔyhbC + ΔelaA. However, for L-Asparaginase which is a secretory protein, the best results were obtained using a combination of ΔelaA+ΔcysW knock-outs. We then tested all the knock outs for their ability to enhance the expression of a 'difficult-to-express' protein. The Rubella virus E1 protein was chosen and tagged with sfGFP at the C-terminal using a linker peptide for easy online monitoring of expression of this fusion protein. Interestingly, the highest increase in Rubella-sGFP levels was obtained in the same double knock-out ΔelaA + ΔcysW (5.6 fold increase in expression yield compared to the control) which gave the highest expression for L-Asparaginase. However, for sfGFP alone, the ΔyhbC+ΔmarR knock-out gave the highest level of expression. These results indicate that there is a fair degree of commonality in the nature of the CSR generated by the induction of different proteins. Transcriptomic profiling of the double knock out showed that many genes associated with the translational machinery and energy biosynthesis did not get down-regulated post induction, unlike the control where these genes were significantly down-regulated. This confirmed our hypothesis of these genes playing an important role in the generation of the CSR and allowed us to design a strategy for making better expression hosts by simply knocking out key genes. This strategy is radically superior to the previous approach of individually up-regulating critical genes since it blocks the mounting of the CSR thus preventing the down-regulation of a very large number of genes responsible for sustaining the flux through the recombinant protein production pathway.

Keywords: cellular stress response, GFP, knock-outs, up-regulated genes

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Authors: Elena K. Schneider, Johnny X. Huang, Vincenzo Carbone, Mark Baker, Mohammad A. K. Azad, Matthew A. Cooper, Jian Li, Tony Velkov

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Ivacaftor is a novel CF trans-membrane conductance regulator (CFTR) potentiator that improves the pulmonary function for cystic fibrosis patients bearing a G551D CFTR-protein mutation. Because ivacaftor is highly bound (>97%) to plasma proteins, there is the strong possibility that co-administered CF drugs that compete for the same plasma protein binding sites and impact the free drug concentration. This in turn could lead to drastic changes in the in vivo efficacy of ivacaftor and therapeutic outcomes. This study compares the binding affinity of ivacaftor and co-administered CF drugs for human serum albumin (HSA) and α1-acid glycoprotein (AGP) using surface plasmon resonance and fluorimetric binding assays that measure the displacement of site selective probes. Due to their high plasma protein binding affinities, drug-drug interactions between ivacaftor are to be expected with ducosate, montelukast, ibuprofen, dicloxacillin, omeprazole and loratadine. The significance of these drug-drug interactions is interpreted in terms of the pharmacodynamic/pharmacokinetic parameters and molecular docking simulations. The translational outcomes of the data are presented as recommendations for a staggered treatment regimen for future clinical trials which aims to maximize the effective free drug concentration and clinical efficacy of ivacaftor.

Keywords: human α-1-acid glycoprotein, binding affinity, human serum albumin, ivacaftor, cystic fibrosis

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Authors: Norhanizan U., S. H. Ahmad, N. A. P. Abdullah, G. B. Saleh

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Physalis minima from the family of Solanaceae is one of the promising fruits which contains the high amount of vitamin C and other antioxidants as well. However, it is a perishable fruit where the deterioration process will commence if the fruits are not stored in proper conditions. There is not much work has been carried out to study the effects of storage durations on Physalis fruit. Therefore, this study was conducted to determine the effects of 0, 3, 6, and 9 days of storage on postharvest quality of Physalis minima fruits. Total of 120g of uniform sizes of fruits (2.3 to 2.5g) were used for each replication and the experiment was repeated thrice. The fruits were divided equally into four groups with each group labeled according to the days of storage. The fruits were then stored in the cool room for nine days with temperature maintain at 12 ° C. The fruits were analyzed for weight loss, firmness, color (L*, C* and hue angle), titratable acidity (TA), soluble solids concentrations (SSC), pH and ascorbic acids. Data were analyzed using analysis of variance and means was separated using least significant difference (LSD). The storage durations affect the quality characteristics of the fruits. On the day 9, the average of fruit weight loss and fruit firmness decreased about 21 and 24% respectively. The level of ascorbic acids and titrable acidity were also decreased while the soluble solids concentration increased during storage. Thus, in order to retain the quality of the fruits, it is recommended that the Physalis fruit can be stored only up to 6 days at 12 ° C.

Keywords: fruit quality, Physalis minima, Solanaceae, storage durations

Procedia PDF Downloads 277

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: Sanxiong Fu, Yanyan Lv, Song Chen, Wei Zhang, Cunkou Qi

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Ethylene response factor proteins are known to play an important role in regulating a variety of stress responses in plants, but their exact functions in submergence stress are not completely understood. In this study, we isolated BnERF from Brassica napus L. to study the function of BnERF in submergence tolerance. The expression of BnERF gene in Brassica napus L. and the expression of antioxidant enzyme genes in transgenic Arabidopsis were analyzed by Quantitative RT-PCR. It was found that expression of BnERF is apparently induced by submergence in Brassica napus L. and overexpression of BnERF in Arabidopsis increases the tolerance level to submergence and oxidative stress. Histochemical method detected lower level of H2O2, O2•− and malondialdehyde (MDA) in the transgenic Arabidopsis. Compared to wild type, transgenic lines also have higher soluble sugar content and higher activity of antioxidant enzymes, which helps protect the plants against the oxidative damage caused by submergence. It was concluded that BnERF can increase the tolerance of plants to submergence stress and BnERF might be involved in regulating soluble sugar content and the antioxidant system in the defense against submergence stress.

Keywords: antioxidant enzyme, Arabidopsis, ethylene response factor, submergence

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Authors: Tagouelbe Tiho, Amissa Augustin Adima, Yao Casimir Brou, Nabayo Traore, Gouha Firmin Kouassi, Thierry Roland Kouame, Maryline Kouba

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The work aimed to study Borassus aethiopum Mart (B.a) dried pulp nutritional value for its incorporation in human and poultry diets. Firstly, the mature fruit macro-composition was assessed. Secondly, the pulp was dried at 40, 50, 60, 70, and 80ᵒC. Thereafter, the analysis was performed for fat, protein, total sugars, Ca, P, Mg, and fatty acid profile monitoring. As a result, the fruits weighed 1,591.35, delivered 516.73, and 677.82 grams of pulp and seeds, respectively. Mainly, increasing heat adversely affected the outputs. Consequently, the fat results were 14.12, 12.97, 8.93, 8.89ᶜ, and 5.56%; protein contents were 11.64, 10.15, 8.97, 8.84, and 8.42%; total sugar deliveries were 6.28, 6.05, 5.26, 5.02, and 4.76% (P < 0.01). Thereafter, the metabolizable energies were 3,785.22; 3,834.28; 3,616.62; 3,667.03; and 3,608.33 kcal/kg (DM). Additionally, Calcium (Ca) contents were 0.51, 0.55, 0.69, 0.77, and 0.81%, while phosphorus (P) mean was 0.17%, and the differences were not significant (P < 0.01). So, the Ca/P ratios were 2.79, 3.04, 4.10, 4.71, and 4.95. Finally, fatty acids (FA) assessments revealed 22.33 saturated (SFA), 77.67 unsaturated (UFA), within which 67.59% were monounsaturated (MUFA). Interestingly, the rising heat depressed n-6/n-3 ratios that were 1.1, 1.1, 0.45 and 0.38, respectively at 40, 50, 70 and 80ᵒC. In short, drying did not only enhance the product shelf life but it also improved the nutritional value. Thus, B.a mature fruit pulps dried at 70ᵒC are good functional foods, with more than 66% MUFA, and energy source for human and poultry nutrition.

Keywords: Borassus aethiopum Mart, fatty acids, metabolizable energy, minerals, protein

Procedia PDF Downloads 164

Authors: Hamad Rafique

Abstract:

The bioactive peptide RDFPITWPW (RW-9) identified from oat protein has been reported to be positive in memory deficits. However, no clarity on the mechanisms responsible for the neuroprotective effects of RW-9 peptide against AD-like symptoms. Herein, it found that RW-9 intervention showed various improving effects in cognitive-behavioral tests and alleviated oxidative stress and inflammation in the scopolamine-induced mice model. The hippocampus proteomics analysis revealed the upregulation of memory-related proteins, including Grin3a, Ppp2r1b, Stat6, Pik3cd, Slc5a7, Chrm2, mainly involved in cAMP signaling, PI3K-Akt signaling, and JAK-STAT signaling pathways. The administration of RW-9 significantly upregulated the neurotransmitters, including 5-HT, DA, and Arg, in mice brains. Moreover, it regulated the serum metabolic profile and increased the expression levels of ABC transporters, biosynthesis of amino acids, and Amino acyl-tRNA biosynthesis, among others. The 16s-rRNA results illustrated that the RW-9 restored the abundance of Muribaculaceae, Lachnospiraceae, Lactobacillus, Clostridia and Bactericides. Taken together, our results suggest that the RW-9 may prevent the AD-like symptoms via modulation of the gut-serum-brain axis.

Keywords: oat protein, active peptide, neuroprotective, gut-brain axis

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Authors: M. Michael Gromiha, A. Mary Thangakani, Sandeep Kumar, D. Velmurugan

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The main cause of several neurodegenerative diseases such as Alzhemier, Parkinson, and spongiform encephalopathies is formation of amyloid fibrils and plaques in proteins. We have analyzed different sets of proteins and peptides to understand the influence of sequence-based features on protein aggregation process. The comparison of 373 pairs of homologous mesophilic and thermophilic proteins showed that aggregation-prone regions (APRs) are present in both. But, the thermophilic protein monomers show greater ability to ‘stow away’ the APRs in their hydrophobic cores and protect them from solvent exposure. The comparison of amyloid forming and amorphous b-aggregating hexapeptides suggested distinct preferences for specific residues at the six positions as well as all possible combinations of nine residue pairs. The compositions of residues at different positions and residue pairs have been converted into energy potentials and utilized for distinguishing between amyloid forming and amorphous b-aggregating peptides. Our method could correctly identify the amyloid forming peptides at an accuracy of 95-100% in different datasets of peptides.

Keywords: aggregation, amyloids, thermophilic proteins, amino acid residues, machine learning techniques

Procedia PDF Downloads 608