Search results for: chick bean protein isolate

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: Supawadee Phetkhajone, Wisuwat Songnuan

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Metalaxyl is one of the most common and effective fungicides used to control Phytophthora palmivora infection in durian (Durio zibethinus L.). The efficacy of metalaxyl residue in durian under greenhouse condition was evaluated using bioassay. Durian seedlings were treated with 2 methods of application, spraying, and soil drenching of metalaxyl, at recommended concentration (1000 mg/L). Mock treated samples were treated with 0.1% Tween20 and water for spraying and soil drenching methods, respectively. The experiment was performed in triplicates. Leaves were detached from treated plants at 0, 1, 7, 15, 20, 30, and 60 days after application, inoculated with metalaxyl-resistant and metalaxyl-sensitive isolates of P. palmivora, and incubated in a high humidity chamber for 5 days at room temperature. Metalaxyl efficacy was determined by measuring the lesion size on metalaxyl treated and mock treated samples. The results showed that metalaxyl can control metalaxyl-sensitive isolate of P. palmivora for at least 30 days after application in both methods of application. The metalaxyl-resistant isolate was not inhibited in all treatments. Leaf samples from spraying method showed larger lesions compared to soil drench method. These results demonstrated that metalaxyl applications, especially soil drenching methods showed high efficacy to control metalaxyl-sensitive isolates of P. palmivora, although it cannot control metalaxyl-resistant isolates of P. palmivora in all treatments. These qualitative data indicate that metalaxyl may suitable to control metalaxyl-sensitive isolates of P. palmivora infection.

Keywords: bioassay, degradation, durian, metalaxyl

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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: 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: Ayobami Solomon Popoola, Victor N. Enujiugha

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Production of soy-yogurt by fermentation of soymilk with lactic acid bacteria isolated from soymilk was studied. Soymilk was extracted from dehulled soybean seeds and pasteurized at 95 °C for 15 min. The soymilk was left to naturally ferment (temperature 40 °C; time 8 h) and lactic acid bacteria were isolated, screened and selected for yogurt production. Freshly prepared soymilk was pasteurized (95 °C, 15 min), inoculated with the lactic acid bacteria isolated (3% w/v starter culture) and incubated at 40 °C for 8 h. The yogurt produced was stored at 4 °C. Investigations were carried out with the aim of improving the sensory qualities and acceptability of soy yogurt. Commercial yogurt was used as a control. The percentage of soymilk inoculated was 70% of the broth. Soy-yoghurt samples produced were subsequently subjected to biochemical and microbiological assays which included total viable counts of fresh milk and soy-based yoghurt; proximate composition of functional soy-based yoghurt fermented with Lactobacillus plantarum; changes in pH, Titratable acidity, and lactic acid bacteria during a 14 day period of storage; as well as morphological and biochemical characteristics of lactic acid bacteria isolated. The results demonstrated that using Lactobacillus plantarum to inoculate soy milk for yogurt production takes about 8 h. The overall acceptability of the soy-based yogurt produced was not significantly different from that of the control sample. The use of isolate from soymilk had the added advantage of reducing the cost of yogurt starter culture, thereby making soy-yogurt, a good source of much desired good quality protein. However, more experiments are needed to improve the sensory qualities such as beany or astringent flavor and color.

Keywords: soy, soymilk, yoghurt, starter culture

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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: Jong Il Rhee

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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: 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: Eleni C. Mazarakioti, Anastasios Zotos, Anna-Akrivi Thomatou, Efthimios Kokkotos, Achilleas Kontogeorgos, Athanasios Ladavos, Angelos Patakas

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“Gigantes Elefantes” is a particularly dynamic crop of giant beans cultivated in western Macedonia (Greece). This variety of large beans growing in this area and specifically in the regions of Prespes and Kastoria is a protected designation of origin (PDO) species with high nutritional quality. Mislabeling of geographical origin and blending with unidentified samples are common fraudulent practices in Greek food market with financial and possible health consequences. In the last decades, multi-elemental composition analysis has been used in identifying the geographical origin of foods and agricultural products. In an attempt to discriminate the authenticity of Greek beans, multi-elemental analysis (Ag, Al, As, B, Ba, Be, Ca, Cd, Co, Cr, Cs, Cu, Fe, Ga, Ge, K, Li, Mg, Mn, Mo, Na, Nb, Ni, P, Pb, Rb, Re, Se, Sr, Ta, Ti, Tl, U, V, W, Zn, Zr) was performed by inductively coupled plasma mass spectrometry (ICP-MS) on 320 samples of beans, originated from Greece (Prespes and Kastoria), China and Poland. All samples were collected during the autumn of 2021. The obtained data were analysed by principal component analysis (PCA), an unsupervised statistical method, which allows for to reduce of the dimensionality of the enormous datasets. Statistical analysis revealed a clear separation of beans that had been cultivated in Greece compared with those from China and Poland. An adequate discrimination of geographical origin between bean samples originating from the two Greece regions, Prespes and Kastoria, was also evident. Our results suggest that multi-elemental analysis combined with the appropriate multivariate statistical method could be a useful tool for bean’s geographical authentication. Acknowledgment: This research has been financed by the Public Investment Programme/General Secretariat for Research and Innovation, under the call “YPOERGO 3, code 2018SE01300000: project title: ‘Elaboration and implementation of methodology for authenticity and geographical origin assessment of agricultural products.

Keywords: geographical origin, authenticity, multi-elemental analysis, beans, ICP-MS, PCA

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Authors: Amal Saafan, Kareem Al Sofy, Sameh AbdelGhani, Magdy Amin

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Background: Acinetobacter spp. resistance towards β-lactam antibiotics is mediated mainly by different classes of β-lactamases production; detection of some genes responsible for production of β-lactamases is the objective of the study. Methods: One hundred fifty bacterial isolates were recovered from blood, sputum, and urine specimens from different hospitals in Egypt. Sixty-nine isolate were identified as Acinetobacter baumannii using traditional biochemical tests, CHROM agar, MicroScan and PCR amplification of blaoxa-51like gene. Acinetobacterbaumannii isolates were grouped into carbapenem resistant group (GP1), cefotaxime, ceftazidime and cefoxitin resistant group (GP2) and carbapenem and cephalosporin non-resistant group (GP3). Carbapenemase activity was screened using modified Hodge test (MHT) for GP1.Metallo-β-lactamases screening was performed for MHT positive isolates using double disk synergy test (DDST) and combined disk test (CDT). Amp C activity was screened using Amp C disk test with Tris-EDTA, DDST, and CDT for GP2. Finally, PCR amplification of blaoxa-51like, blaoxa-23like, blaIMP-like, blaVIM-like, and blaADC-like genes was performed for isolates that showed, at least, two positive results of three for both AmpC and carbapenemases phenotypic screening tests (obvious activity), in addition to GP3 (for comparison). Detection of blaoxa-51like and blaADC-like genes preceded by ISAba1 was also performed. Results: Antibiogram of 69 pure Acinetobacter baumannii isolates resulted in 57, 64, and 2 isolates enrolled into GP1, GP2, and GP3, respectively. Carbapenemase activity was shown by 49(85.9%) isolate using MHT. Metallo-β-lactamases screening revealed 32(65.3%) and 35(71.4%) using DDST and CDT, respectively.AmpC activity was shown by 43(67.2%) and 50 (78.1%) isolates using AmpC disk test with Tris-EDTA, and both DDST and CDT, respectively. Twenty-seven isolates showed obvious activity, all of them (100%) were harboring blaoxa-51like and blaADC-like genes, while blaoxa-23like, blaIMP-like andblaVIM-like genes were harbored by 23(85.2%), 9 (33.%) and no isolate respectively. Only 12 (44.4%) isolates harbored blaoxa-51like and blaADC-like genes preceded by ISAba1. GP3 isolates showed only positive blaoxa-51like and blaADC-like genes. Conclusion: It is not possible to correlate resistance with presence of blaoxa-51like and blaADC-like genes and presence of ISAba1 was immediate as transcriptional promoter. A blaoxa-23like gene played an important role in carbapenem resistance when compared with blaIMP-like and blaVIM-like gene.

Keywords: acinetobacter, beta-lactams, resistance, antimicrobial agents

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Authors: Ashima Sharma

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Human Serum Albumin (HSA)- one of the most demanded therapeutic proteins with immense biotechnological applications- is a large multidomain protein containing 17 disulfide bonds. The current source of HSA is human blood plasma which is a limited and unsafe source. Thus, there exists an indispensable need to promote non-animal derived recombinant HSA (rHSA) production. Escherichia coli is one of the most convenient hosts which had contributed to the production of more than 30% of the FDA approved recombinant pharmaceuticals. It grows rapidly and reaches high cell density using inexpensive and simple substrates. E. coli derived recombinant products have more economic potential as fermentation processes are cheaper compared to the other expression hosts. The major bottleneck in exploiting E. coli as a host for a disulfide-rich multidomain protein is the formation of aggregates of overexpressed protein. The majority of the expressed HSA forms inclusion bodies (more than 90% of the total expressed rHSA) in the E. coli cytosol. Recovery of functional rHSA from inclusion bodies is not preferred because it is difficult to obtain a large multidomain disulfide bond rich protein like rHSA in its functional native form. Purification is tedious, time-consuming, laborious and expensive. Because of such limitations, the E. coli host system was neglected for rHSA production for the past few decades despite its numerous advantages. In the present work, we have exploited the capabilities of E. coli as a host for the enhanced functional production of rHSA (~60% of the total expressed rHSA in the soluble fraction). Parameters like intracellular environment, temperature, induction type, duration of induction, cell lysis conditions etc. which play an important role in enhancing the level of production of the desired protein in its native form in vivo have been optimized. We have studied the effect of assistance of different types of exogenously employed chaperone systems on the functional expression of rHSA in the E. coli host system. Different aspects of cell growth parameters during the production of rHSA in presence and absence of molecular chaperones in E. coli have also been studied. Upon overcoming the difficulties to produce functional rHSA in E. coli, it has been possible to produce significant levels of functional protein through engineering the biological system of protein folding in the cell, the E. coli-derived rHSA has been purified to homogeneity. Its detailed physicochemical characterization has been performed by monitoring its conformational properties, secondary and tertiary structure elements, surface properties, ligand binding properties, stability issues etc. These parameters of the recombinant protein have been compared with the naturally occurring protein from the human source. The outcome of the comparison reveals that the recombinant protein resembles exactly the same as the natural one. Hence, we propose that the E. coli-derived rHSA is an ideal biosimilar for human blood plasma-derived serum albumin. Therefore, in the present study, we have introduced and promoted the E. coli- derived rHSA as an alternative to the preparation from a human source, pHSA.

Keywords: recombinant human serum albumin, Escherichia coli, biosimilar, chaperone assisted protein folding

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Authors: Chaoran Liu

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Bioactive polysaccharides and polysaccharide-protein complex derived from mushrooms and fungi have a wide range of immunomodulatory activity with low side-effects and have therefore the potential to be developed as an adjuvant in cancer therapies. Mushrooms sclerotium is rich in polysaccharides and the polysaccharides isolated from the sclerotium of Polyporus rhinocerus have shown potent in vivo and in vitro immunomodulatory effects. Macrophages are considered to be an important component of the innate immune response against bacterial infection and cancer. To better understanding the immunomodulatory effects and its underlying mechanisms of sclerotial water-soluble polysaccharides extracted from P. rhinocerus on macrophages, the objectives of this study are to purify the water-soluble novel sclerotial polysaccharides and to characterize the structure and properties as well as to study the detailed molecular mechanisms of the in vitro immunomodulating effects in murine macrophages. The hot water-soluble fraction PRW from the sclerotium of P. rhinocerus was obtained using solvent extraction. PRW was further fractionated by membrane ultrafiltration to a give a fraction (PRW1) with molecular mass less than 50 kDa. PRW1 was characterized to be a polysaccharide-protein complex composed of 45.7% polysaccharide and 44.2% protein. The chemical structure of the carbohydrate moiety of PRW1 was elucidated by GC and FTIR to be mainly beta-D-glucan with trace amount of galactose and mannose. The immunomodulatory effects of PRW1 on murine RAW 264.7 macrophages were demonstrated in terms of the increase in nitric oxide production and cytokine production. Mechanistically, PRW1 initiates ERK phosphorylation to activate macrophages within 15 min and significantly improves the expression level of inducible NOS (iNOS) from 6 h after treatment. In summary, this study indicates that PRW1 is a potent immunomodulatory agent for macrophages and suggests that mushroom sclerotia from Polyporus rhinocerus requires for further investigation in cancer research.

Keywords: Polyporus rhinocerus, mushroom sclerotia, Polysaccharide-Protein Complex, macrophage activation

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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

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Authors: Hamad Rafique

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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 615

Authors: Srijata Mitra, Mobina Parveen, Pranab Roy, Narayan Chandra Chattopadhyay

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Chlorinated hydrocarbon can be a major pollution problem in groundwater as well as soil. Many people interact with these chemicals on daily accidentally or by professionally in the laboratory. One of the most common sources for Chlorinated hydrocarbon contamination of soil and groundwater are industrial effluents. The wide use and discharge of Trichloroethylene (TCE), a volatile chlorohydrocarbon from chemical industry, led to major water pollution in rural areas. TCE is an mainly used as an industrial metal degreaser in industries. Biotransformation of TCE to the potent carcinogen vinyl chloride (VC) by consortia of anaerobic bacteria might have role for the above purpose. For these reasons, the aim of current study was to isolate and characterized the genes involved in TCE metabolism and also to investigate the in silico study of those genes. To our knowledge, only one aromatic dioxygenase system, the toluene dioxygenase in Pseudomonas putida F1 has been shown to be involved in TCE degradation. This is first instance where Bacillus cereus group being used in biodegradation of trichloroethylene. A novel bacterial strain 2479 was isolated from oil depot site at Rajbandh, Durgapur (West Bengal, India) by enrichment culture technique. It was identified based on polyphasic approach and ribotyping. The bacterium was gram positive, rod shaped, endospore forming and capable of degrading trichloroethylene as the sole carbon source. On the basis of phylogenetic data and Fatty Acid Methyl Ester Analysis, strain 2479 should be placed within the genus Bacillus and species cereus. However, the present isolate (strain 2479) is unique and sharply different from the usual Bacillus strains in its biodegrading nature. Fujiwara test was done to estimate that the strain 2479 could degrade TCE efficiently. The gene for TCE biodegradation was PCR amplified from genomic DNA of Bacillus cereus 2479 by using todC1 gene specific primers. The 600bp amplicon was cloned into expression vector pUC I8 in the E. coli host XL1-Blue and expressed under the control of lac promoter and nucleotide sequence was determined. The gene sequence was deposited at NCBI under the Accession no. GU183105. In Silico approach involved predicting the physico-chemical properties of deduced Tce1 protein by using ProtParam tool. The tce1 gene contained 342 bp long ORF encoding 114 amino acids with a predicted molecular weight 12.6 kDa and the theoretical pI value of the polypeptide was 5.17, molecular formula: C559H886N152O165S8, total number of atoms: 1770, aliphatic index: 101.93, instability index: 28.60, Grand Average of Hydropathicity (GRAVY): 0.152. Three differentially expressed proteins (97.1, 40 and 30 kDa) were directly involved in TCE biodegradation, found to react immunologically to the antibodies raised against TCE inducible proteins in Western blot analysis. The present study suggested that cloned gene product (TCE1) was capable of degrading TCE as verified chemically.

Keywords: cloning, Bacillus cereus, in silico analysis, TCE

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Authors: Natela Gerliani, Mohammed Aider

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Nowadays, the interest of using sustainable technologies for protein extraction from underutilized oilseeds is growing. Currently, a major disposal problem for the oil industry is by-products of plant food processing such as soybean meal. That is why valorization of soybean meal is important for the oil industry since it contains high-quality proteins and other valuable components. Generally, soybean meal is used in livestock and poultry feed but is rarely used in human feed. Though chemical composition of this meal compensate nutritional deficiency and can be used to balance protein in human food. Regarding the efficiency of soybean meal valorization, extraction is a key process for obtaining enriched protein ingredient, which can be incorporated into the food matrix. However, most of the food components such as proteins extracted from oilseeds by-products imply the utilization of organic and inorganic chemicals (e.g. acids, bases, TCA-acetone) having a significant environmental impact. In a context of sustainable production, the use of an electro-activation technology seems to be a good alternative. Indeed, the electro-activation technology requires only water, food grade salt and electricity as main materials. Moreover, this innovative technology helps to avoid special equipment and trainings for workers safety as well as transport and storage of hazardous materials. Electro-activation is a technology based on applied electrochemistry for the generation of acidic and alkaline solutions on the basis of the oxidation-reduction reactions that occur at the vicinity electrode/solution interfaces. It is an eco-friendly process that can be used to replace the conventional acidic and alkaline extraction. In this research, the electro-activation technology for protein extraction from soybean meal was carried out in the electro-activation reactor. This reactor consists of three compartments separated by cation and anion exchange membranes that allow creating non-contacting acidic and basic solutions. Different current intensities (150 mA, 300 mA and 450 mA) and treatment durations (10 min, 30 min and 50 min) were tested. The results showed that the extracts obtained by the electro-activation method have good quality in comparison to conventional extracts. For instance, extractability obtained with electro-activation method was 55% whereas with the conventional method it was only 36%. Moreover, a maximum protein quantity of 48 % in the extract was obtained with the electro-activation technology comparing to the maximum amount of protein obtained by conventional extraction of 41 %. Hence, the environmentally sustainable electro-activation technology seems to be a promising type of protein extraction that can replace conventional extraction technology.

Keywords: by-products, eco-friendly technology, electro-activation, soybean meal

Procedia PDF Downloads 229

Authors: Abdu Umar Adamu, Saidu Akun Abdullahi, Al-Hassan Muhammed, Hamisu Abdu

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Leguminous plants such as beans have been considered as a source of protein in this present work. The proximate analysis on beans (Vigna unguiculata) were determined in order to identify the nutritional content as well as presence of some heavy metals accumulation in washed and unwashed beans (white Beans) sold in Kazaure market Jigawa State Nigeria. On the average comparative analysis, the result has indicated that, the Vigna unguiculata had protein content of 61.1%, fibre 4.5%, ash 10.4%, moisture 5%, carbohydrate 15.8% and total lipid 4.9%, therefore it could be suggested that beans has enough nutritional content that helps the people health. The heavy metal analysis of unwashed white beans showed that Fe (17.37 ± 6.71)mg/kg had the highest concentration followed by Zn (6.41 ± 3.09), Cu (5.69 ± 2.42), Cd (0.46 ± 0.65) and Pb (0.57 ± 0.94)mg/kg , while the washed beans shows that Zn (0.11 ± 0.17), Fe (0.01 ± 0.006), Cd (0.02 ± 0.01), Cu (0.03 ± 0.021), Pb (0.01 ± 0.006)mg/kg. The washed white beans are safe for consumption and also the concentration of heavy metal are negligible and of nontoxic effect to human health.

Keywords: white beans, protein, proximate composition, heavy metal

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Authors: Ruolan Zhou

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Trisomy of human chromosome 21 is the primary cause of Down Syndrome (DS), and this genetic disease has significantly burdened families and countries, causing great controversy. To address this problem, the research takes an approach in exploring the relationship between genetic abnormality and this disease's symptoms, adopting several techniques, including data analysis and enrichment analysis. It also explores open-source websites, such as NCBI, DAVID, SOURCE, STRING, as well as UCSC, to complement its result. This research has analyzed the variety of genes on human chromosome 21 with simple coding, and by using analysis, it has specified the protein-coding genes, their function, and their location. By using enrichment analysis, this paper has found the abundance of keratin production-related coding-proteins on human chromosome 21. By adopting past researches, this research has attempted to disclose the relationship between trisomy of human chromosome 21 and keratin production abnormality, which might be the reason for common diseases in patients with Down Syndrome. At last, by addressing the advantage and insufficiency of this research, the discussion has provided specific directions for future research.

Keywords: Down Syndrome, protein production, genome, enrichment analysis

Procedia PDF Downloads 127

Authors: Anjum-Zubair, Muhammad, Muhammad Shoaib Alam, Muhammad Samee Mubarik, Iftikhar Ahmad

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The aim of present research was to evaluate the effect of dietary protein (soybean) formulated feed on the growth performance of carp fish seed (Rohu, Mori, Grass, and Gulfam) in ponds under polyculture system. Keeping in view the protein requirements of these four carps, they were fed with formulated feed contains 30% of crude protein. The fingerlings were fed once on daily basis at 5% of their wet body weight. A 90 days experiment was conducted in two cemented ponds situated at Fish Seed Hatchery and Research Centre, Rawal Town, Islamabad, Pakistan. Pond1 contain major carps i.e. Rohu and Mori while pond 2 was stocked with Chinese carps i.e. Grass carp and Gulfam. Random sampling of five individuals of each species was done fortnightly to measure the body weight and total body length. Maximum growth was observed in fingerling of Grass carp followed by Mori, Rohu and Gulfam. Total fish production was recorded as Grass 623.45 gm followed by Mori 260.3 gm, Rohu 243.08 gm and Gulfam 181.165 gm respectively. Significantly results were obtained among these four fish species when the corresponding data was subjected to statistical analysis by using two sample t-test. The survival rate was 100%. Study shows that soybean as plant based protein can be easily used as substitute to fish meal without any adverse effect on fish health and fish production.

Keywords: carps, fry growth, poly culture, soybean meal

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Authors: L. A. Bosnea, T. Moschakis, C. Biliaderis

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Two probiotic strains of Lactobacillus paracasei subsp. paracasei (E6) and Lactobacillus paraplantarum (B1), isolated from traditional Greek dairy products, were microencapsulated by complex coacervation using whey protein isolate (WPI, 3% w/v) and gum arabic (GA, 3% w/v) solutions mixed at different polymer ratio (1:1, 2:1 and 4:1). The effect of total biopolymer concentration on cell viability was assessed using WPI and GA solutions of 1, 3 and 6% w/v at a constant ratio of 2:1. Also, several parameters were examined for optimization of the microcapsule formation, such as inoculum concentration and the effect of ionic strength. The viability of the bacterial cells during heat treatment and under simulated gut conditions was also evaluated. Among the different WPI/GA weight ratios tested (1:1, 2:1, and 4:1), the highest survival rate was observed for the coacervate structures made with the ratio of 2:1. The protection efficiency at low pH values is influenced by both concentration and the ratio of the added biopolymers. Moreover, the inoculum concentration seems to affect the efficiency of microcapsules to entrap the bacterial cells since an optimum level was noted at less than 8 log cfu/ml. Generally, entrapment of lactobacilli in the complex coacervate structure enhanced the viability of the microorganisms when exposed to a low pH environment (pH 2.0). Both encapsulated strains retained high viability in simulated gastric juice (>73%), especially in comparison with non-encapsulated (free) cells (<19%). The encapsulated lactobacilli also exhibited enhanced viability after 10–30 min of heat treatment (65oC) as well as at different NaCl concentrations (pH 4.0). Overall, the results of this study suggest that complex coacervation with WPI/GA has a potential to deliver live probiotics in low pH food systems and fermented dairy products; the complexes can dissolve at pH 7.0 (gut environment), releasing the microbial cells.

Keywords: probiotic, complex coacervation, whey, encapsulation

Procedia PDF Downloads 298

Authors: F. Bellali, M. Kharroubi, M. Loutfi, N.Bourhim

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In Morocco, fish processing industry is an important source income for a large amount of byproducts including skins, bones, heads, guts and scales. Those underutilized resources particularly scales contain a large amount of proteins and calcium. Scales from Sardina plichardus resulting from the transformation operation have the potential to be used as raw material for the collagen production. Taking into account this strong expectation of the regional fish industry, scales sardine upgrading is well justified. In addition, political and societal demands for sustainability and environment-friendly industrial production systems, coupled with the depletion of fish resources, drive this trend forward. Therefore, fish scale used as a potential source to isolate collagen has a wide large of applications in food, cosmetic and bio medical industry. The main aim of this study is to isolate and characterize the acid solubilize collagen from sardine fish scale, Sardina pilchardus. Experimental design methodology was adopted in collagen processing for extracting optimization. The first stage of this work is to investigate the optimization conditions of the sardine scale deproteinization on using response surface methodology (RSM). The second part focus on the demineralization with HCl solution or EDTA. Moreover, the last one is to establish the optimum condition for the isolation of collagen from fish scale by solvent extraction. The basic principle of RSM is to determinate model equations that describe interrelations between the independent variables and the dependent variables.

Keywords: Sardina pilchardus, scales, valorization, collagen extraction, response surface methodology

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Authors: A. Ojha, Y. K. Gupta

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Organophosphate (OP) pesticides are among the most widely used synthetic chemicals for controlling a wide variety of pests throughout the world. Chlorpyrifos (CPF), methyl parathion (MPT), and malathion (MLT) are among the most extensively used OP pesticides in India. DNA strand breaks and DNA-protein crosslinks (DPC) are toxic lesions associated with the mechanisms of toxicity of genotoxic compounds. In the present study, we have examined the potential of CPF, MPT, and MLT individually and in combination, to cause DNA strand breakage and DPC formation. Peripheral blood lymphocytes of rat were exposed to 1/4 and 1/10 LC50 dose of CPF, MPT, and MLT for 2, 4, 8, and 12h. The DNA strand break was measured by the comet assay and expressed as DNA damage index while DPC estimation was done by fluorescence emission. There was significantly marked increase in DNA damage and DNA-protein crosslink formation in time and dose dependent manner. It was also observed that MPT caused the highest level of DNA damage as compared to other studied OP compounds. Thus, from present study, we can conclude that studied pesticides have genotoxic potential. The pesticides mixture does not potentiate the toxicity of each other. Nonetheless, additional in vivo data are required before a definitive conclusion can be drawn regarding hazard prediction to humans.

Keywords: organophosphate, pesticides, DNA damage, DNA protein crosslink, genotoxic

Procedia PDF Downloads 356

Authors: Izzatul Ulfana, Angga Pratomo Cahyadi, Rimayanti, Widjiati

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Freezing oocyte could cause temperature stress. Temperature stress triggers cell damage. Insulin Transferrin Selenium (ITS) and Heat Shock Protein 70 (HSP70) had been used to prevent damage to the oocyte after freezing. ITS and HSP70 could cause the difference protective effect. The aim of this research was to obtain an effective cryoprotectant for freezing local goat oocyte in cumulus cells change. The research began by collecting the ovary from a local slaughterhouse in Indonesia, aspiration follicle, in vitro maturation and the freezing had been used vitrification method. Examination of the morphology cells by native staining method. Data on the calculation morphology oocyte analyzed by Kruskall-Wallis Test. After the Kruskall-Wallis Test which indicated significance, followed by Mann-Whitney Test to compare between treatment groups. As a result, cryoprotectant ITS has the best culumus cells after warming

Keywords: Insulin Transferrin Selenium, Heat Shock Protein 70, cryoprotectant, vitrification

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Authors: Jiraporn Rojtinnakorn

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ICAM-2 (intercellular adhesion molecule 2) or CD102 (Cluster of Differentiation 102) is type I trans-membrane glycoproteins, composing 2-9 immunoglobulin-like C2-type domains. ICAM-2 plays the particular role in immune response and cell surveillance. It is concerned in innate and specific immunity, cell survival signal, apoptosis, and anticancer. EST clone of ICAM-2, from P. gigas blood cell EST libraries, showed high identity to human ICAM-2 (92%) with conserve region of ICAM N-terminal domain and part of Ig superfamily. Gene and protein of ICAM-2 has been founded in mammals. This is the first report of ICAM-2 in fish.

Keywords: ICAM-2, CD102, Pangasianodon gigas, antitumor

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Authors: A. Belay, B. Kebede

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The concentration- and temperature-dependent refractive indices of solutions extracted from defective and non-defective coffee beans have been investigated using a He–Ne laser. The refractive index has a linear relationship with the presumed concentration of the coffee solutions in the range of 0.5–3%. Higher and lower values of refractive index were obtained for immature and non-defective coffee beans, respectively. The Refractive index of bean extracts can be successfully used to separate defective from non-defective beans.

Keywords: coffee extract, refractive index, temperature dependence

Procedia PDF Downloads 150

Authors: Ishaq Kashif, Muhammad Younas, Muhammad Riaz, Mubarak Ali

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Poor feeding management during pre-weaning period is one of the factors resulting in compromised growth of Beetal kids fattened for meat purpose. The main reason for this anomaly may be less milk offered to kids and non-serious efforts for its management. This study was planned to find the most appropriate protein level suiting the age of the weaning while shifting animals to high input feeding system. Total of 42 Beetal male kids having 30 (±10), 60 (±10) and 90 (±10) days of age were selected with 16 in each age group. They were designated as G30, G60 and G90, respectively. The weights of animals were; 8±2 kg (G30), 12±2 kg (G60) and 16±2 kg (G90), respectively. All animals were weaned by introducing the total mix feed gradually and withdrawing the milk during the adjustment period of two weeks. The pelleted starter ration (total mix feed) with three various dietary protein levels designated as R1 (16% CP), R2 (20% CP) and R3 (26% CP) were introduced. The control group was reared on the fodder (Maize). The starter rations were iso-caloric and were offered for six-week duration. All animals were exposed to treatment using two-factor factorial (3×3) plus control treatment arrangement under completely randomized design. The data were collected on average daily feed intake (ADFI), average daily gain (ADG), gain to intake ratio, Klieber ratio (KR), body measurements and blood metabolites of kids. The data was analyzed using aov function of R-software. The statistical analysis showed that starter feed protein levels and age of weaning had significant interaction for ADG (P < 0.001), KR (P < 0.001), ADFI (P < 0.05) and blood urea nitrogen (P < 0.05) while serum creatinine and feed conversion had non-significant interaction. The trend analysis revealed that ADG had significant quadratic interaction (P < 0.05) within protein levels and age of weaning. It was found that animals weaned at 30 or 60 days, on R2 diet had better ADG (46.8 gm/day and 87.06 gm/day, respectively) weaned at 60 days of age. The animals weaned at 90 days had best ADG (127 gm/day) with R1. It is concluded that animal weaned at 30 or 40 days required 20% CP for better growth performance while animal at 90 days showed better performance with 16% CP.

Keywords: average daily gain, starter protein levels, weaning age, gain to intake ratio

Procedia PDF Downloads 249