ABSTRACT
The ADP ribosylation factor like protein 15 (ARL15) gene encodes for an uncharacterized GTPase associated with rheumatoid arthritis (RA) and other metabolic disorders. Investigation of the structural and functional attributes of ARL15 is important to position the protein as a potential drug target. Using spectroscopy, we demonstrated that ARL15 exhibits properties inherent of GTPases. The Km and Vmax of the enzyme were calculated to be 100 µM and 1.47 µmole/min/µL, respectively. The equilibrium dissociation constant (Kd) of GTP binding with ARL15 was estimated to be about eight-fold higher than that of GDP. Small Angle X-ray Scattering (SAXS) data indicated that in solution, the apo state of monomeric ARL15 adopts a shape characterized by a globe of maximum linear dimension (Dmax) of 6.1 nm, and upon binding to GTP or GDP, the vector distribution profile changes to peak-n-tail shoulder with Dmax extended to 7.6 and 7.7 nm, respectively. Structure restoration using a sequence-based template and experimental SAXS data provided the first visual insight revealing that the folded N-terminal in the unbound state of the protein may toggle open upon binding to guanine nucleotides. The conformational dynamics observed in the N-terminal region offer a scope to develop drugs that target this unique GTPase, potentially providing treatments for a range of metabolic disorders.
Subject(s)
Arthritis, Rheumatoid , Metabolic Diseases , Humans , Guanine Nucleotides , Nucleotides/metabolism , Guanine , Scattering, Small Angle , X-Ray Diffraction , ADP-Ribosylation Factors/genetics , ADP-Ribosylation Factors/metabolism , Proteins/metabolism , Guanosine Triphosphate/metabolism , Guanosine DiphosphateABSTRACT
This study investigated the extraction of capsaicin from Capsicum chinense cv Bhut Jolokia using rice bran oil (RBO) as a 'green solvent' via microwave- and ultrasound-mediated techniques (MME and UME) as an alternative to hazardous organic solvents. Extraction conditions were optimised using MME at 180 and 300 W for 2 and 8 min and UME at 300 and 400 W for 10 and 30 min. The maximum capsaicin yield obtained was 12.47 mg/mL, with a corresponding gamma oryzanol content of 8.46 mg/mL. The peroxide value of the extract (capsaicin-infused RBO) was found to be within permissible limits as per Codex Alimentarius specifications. Results strongly suggest the potential of RBO as a novel solvent for extraction of capsaicin. The capsaicin-enriched oil can be used as a functional food ingredient due to its health-promoting properties.
ABSTRACT
Black garlic is rich in brown pigments and Maillard reaction products are known for antioxidant activity and health promoting effects. In the present investigation, we report a facile strategy for fabricating low-fat innovative mayonnaise (IM) using black garlic particles (BGP) as a natural pigment, and a functional ingredient. Whey protein concentrate and high methoxyl pectin at optimized concentrations were utilized for fabricating an IM which served as a control. IM5 and IM10 were ternary composites constituting whey protein, high methoxyl pectin along with BGP (@5 and 10% respectively). The formulation IM10 (BGP @10%) showed high firmness and low spreadability quotient, hence IM5 was taken forward for fabrication for two more variants namely IM-J (using low methoxyl pectin (LMP) from jackfruit peels) and IM-C (LMP from citrus). The effect of BGP and LMP on the functional quality of IM was confirmed through zeta potential, antioxidant activity, textural, rheological, and microscopic evaluation. Fluorescence microscopy confirmed the presence of solid particles over the fat phase of IM, while interaction of pectin and whey proteins was demonstrated through fluorescence emission spectroscopy which clearly displayed stabilization of IM through the formation of Pickering emulsion. Pronounced difference in color and flavor score with BGP established high sensory scores in IM5, IM-J, and IM-C. Rheology supported the stabilizing effects of LMP in IM-J and IM-C in terms of speedy recovery of thixotropy, with recovering storage modulus (G'). Enhanced viscosity of IM-C and IM-J further corroborated the dual effect of LMP and BGP in improving emulsifying and functional quality of IM. Enhanced oxidative stability of IM was established by reduced peroxide and Totox values. Overall our results suggest the promising applications of black garlic as functional ingredient in protein and pectin based Pickering emulsions.
Subject(s)
Garlic , Emulsions/chemistry , Whey Proteins/chemistry , Antioxidants , Emulsifying Agents , Pectins/chemistry , RheologyABSTRACT
Purpose: Leishmaniasis, Chagas disease, and sleeping sickness are caused by protozoa Leishmania donovani, Trypanosoma cruzi, and Trypanosoma brucei, respectively. Platelet activating factor acetyl hydrolase (PAF-AH) is an inflammatory protein implicated in pathogenesis of these three infections, thereby making them attractive drug targets. Methods: PAF-AH sequences were retrieved from UniProt and aligned using Clustal Omega. Homologous models of parasitic proteins were built based on crystal structure of human PAF-AH and validated using PROCHECK server. Volumes of substrate-binding channel were calculated using the ProteinsPlus program. High throughput virtual screening using Glide program in Schrodinger was done with ZINC drug library against parasitic PAF-AH enzymes. Complexes with best hits were energy-minimized and subjected to 100 ns molecular dynamic simulation and analyzed. Results: PAF-AH enzyme sequences from protozoa Leishmania donovani, Trypanosoma cruzi, Trypanosoma brucei, and human have a minimum of 34% sequence similarity with each other. Corresponding structures show a globular conformation consisting of twisted ß-pleated sheets, flanked by α-helices on either side. Catalytic triad of serine-histidine-aspartate is conserved. Substrate-binding channel residues are conserved to an extent, with a lower channel volume in human as compared to target enzymes. Drug screening resulted in identification of three molecules that had better affinities than the substrate to the target enzymes. These molecules fulfill Lipinski's rules for drug likeness and also bind with less affinity to the human counterpart, thereby establishing a high selective index. Conclusion: Structures of PAF-AH from protozoan parasites and humans belong to the same family of enzymes and have a similar three-dimensional fold. However, they show subtle variations in residue composition, secondary structure composition, substrate-binding channel volume, and conformational stability. These differences result in certain specific molecules being potent inhibitors of the target enzymes while simultaneously having weaker binding to human homologue.
ABSTRACT
Parkinson's disease and Schizophrenia fall under low dopamine neurodegenerative and high dopamine psychiatric disorders respectively. Pharmacological interventions to correct mid-brain dopamine concentrations sometimes overshoots the physiological dopamine levels leading to psychosis in Parkinson's disease patients and, extra-pyramidal symptoms in schizophrenia patients. Currently no validated method is available to monitor side effects in such patients, Apolipoprotein E is one of the CSF biomarkers identified in the recent past that shows an inverse relation to mid-brain dopamine concentration. In this study, we have developed s-MARSA for the detection of Apolipoprotein E from ultra-small volume (2 µL) of CSF. s-MARSA exhibits a broad detection range (5 fg mL-1 to 4 µg mL-1) with a better detection limit and could be performed within an hour utilizing only a small volume of CSF sample. The values measured by s-MARSA strongly correlates with the values measured by ELISA. Our method has advantages over ELISA in having a lower detection limit, a broader linear detection range, shorter analysis time, and requiring a low volume of CSF samples. The developed s-MARSA method holds promise for the detection of Apolipoprotein E with clinical utility for monitoring pharmacotherapy of Parkinson's and Schizophrenia patients.
Subject(s)
Magnetite Nanoparticles , Parkinson Disease , Schizophrenia , Humans , Parkinson Disease/diagnosis , Parkinson Disease/drug therapy , Schizophrenia/drug therapy , Dopamine , Apolipoproteins/therapeutic useABSTRACT
Purpose: Spike protein on SARS-CoV-2 virus plays an integral part during infection as cell entry depends on binding of this protein to human ACE2 receptor. Understanding of infectivity by these variants necessitates a comparative structural analysis of complexes of spike protein-receptor binding domain (RBD) of these variants to receptor. Methodology: Wild type SARS-CoV-2 spike protein sequence was retrieved from the UniProt database, and mutations of five variants at receptor binding domain were manually incorporated and aligned using Clustal Omega. Crystal structure complexes of human ACE2 receptor with spike protein RBD domain of SARS-CoV-2 variants of wild type, α, ß, and δ were extracted from the RCSB database. Wild type SARS-CoV-2 complex with receptor was used as template to generate model complexes of receptor with spike protein RBD of γ and omicron variants through WinCoot program. These were energy minimized and validated and molecular dynamic simulation was performed using Desmond simulation program. Results: Mutations are distributed across the entire length of RBD, but the maximum number of mutations are seen at 11 positions within binding interface motifs of six variant sequences. Interface of spike protein RBDs with human ACE2-receptor shows different mix of hydrogen bonded and ionic interactions. Alpha and ß variants have few interactions, while γ and δ variants have higher number of interactions compared to wild type variant. Omicron variant, with 10 polar interactions including two ionic bonds, has the highest binding energy. Conclusion: Different mutations on RBD of spike protein results in varying quantity and quality of interactions, thereby affecting potency of each variant. Variations in binding are due to interactions of mutant residues and induced conformational changes on loops of RBDs. Variants α and ß have a low potency, while, γ, δ, and omicron have a higher potency. These results correlate with viral infectivity and place clinical observations in the right perspective.
ABSTRACT
Ultrafiltration (UF) of buffalo skim milk (BSM) induces changes in its delicate protein-mineral equilibrium. Appling UF causes alteration in chemical composition of UF retentates as a function of protein concentration that adversely affect their physical and rheological properties. Hence, present investigation was targeted to evaluate the changes taking place in heat stability, ζ-potential, particle size, apparent viscosity, pH, turbidity and crossover temperature of storage (G') and loss (Gâ³) modulus of high-protein BSM based UF retentates as a function of homogenization and sodium hydrogen phosphate (SHP) addition. The UF of BSM (heat treated at 85 ± 1 °C for 5 min), significantly increased (P < 0.05) the concentration of protein, fat and minerals, however, it decreased the concentration of lactose and water soluble minerals in UF retentates over BSM. The SHP addition significantly increased (P < 0.05) pH, crossover temperature of G' and Gâ³, ζ-potential, while significantly decreased (P < 0.05) turbidity and particle size in most non-homogenized retentates. Heat coagulation time (HCT) of control and treated UF retentates were at par (P > 0.05) with each other, however, variations were observed in their viscosity values. Rheological behaviour of most of these UF retentates was efficiently described by Bingham model. The correlation among ζ-potential, particle size, apparent viscosity, pH, turbidity, HCT and crossover temperatures G' and Gâ³ of evaluated samples was also established. Overall, this study concluded that 0.5-6% SHP addition in non-homogenized UF retentates, markedly improved their milk protein stability as advocated by higher ζ-potential, G' and Gâ³ crossover temperature values. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s13197-021-05097-2.
ABSTRACT
The juice expelled from carrot, a globally produced root vegetable, leavesbehind carrot pomace (a bio- and horticultural waste) which is potentially rich source of micro-nutrients and carotenoids.However, it is discarded as waste or used as animal feed. It holds potential to be channelized to food chain by a couple of technological interventions. In this regard, present work was aimed at preparing stable emulsion based delivery system for 'green' carotenoids extracted from carrot-pomace in flaxseed oil (a green solvent), and at maximizing the amount of core material so that the resultant emulsion can potentially be used as a source of both carotenoids and omega-3 fatty acid of flaxseed oil origin. The study used natural emulsifier. Preparation of oil-in-water emulsion was optimized using 33 factorial experiment by varying levels of extract containing carotenoid (30-40%), whey protein concentrates (WPC-80) and lactose. The optimized emulsion (CREm) was selected on the basis of particle size, zeta potential, color values (L*, a*, b*) and viscosity statistically analyzed via three-way ANOVA using Proc GLM of SAS 9.3 (described in detail in this paper); the respective values of these parameters being 120.03 ± 8.20 nm, -16.57 ± 0.49 mV, 75.11 ± 0.04, 9.66 ± 0.32, 50.29 ± 0.62, and 0.124 ± 0.0115 Pa.s for CREm. CREm contained 35% flaxseed oil, 10% WPC-80 and 5% lactose and showed good centrifugal and gravitational stability (15 days). It was analyzed for total carotenoid content, antioxidant activities (ABTS (2,2-azinobis-(3-ethylbenzthiazoline-6sulfonic acid), DPPH (2,2-Diphenyl-1-picrylhydrazyl) and FRAP (Ferric reducing antioxidant power assay)) and microstructure.
Subject(s)
Daucus carota , Analysis of Variance , Animals , Carotenoids , Emulsions , Linseed OilABSTRACT
The bio-wastes (like peels, seeds, etc.) from food industry are rich source of bio-active components, but are poorly managed. In present study, carotenoids were extracted from carrot pomace using ultrasonication and high shear dispersion techniques and flaxseed oil as green solvent (green biorefinery approach). Various combinations of time and temperature were used and final selection was made on the basis of maximum recovery of carotenoids. High shear disperser yielded maximum carotenoids (recovery 94.8 ± 0.08%). The total carotenoid content, antioxidant activity as ABTS, DDPH and FRAP and ß-carotene of carotenoid rich extract from carrot pomace (CREP) were 82.66 ± 0.06 µg/g, 1596.04 ± 69.45 µg Trolox eq./ml, 380.21 ± 39.62 µg Trolox eq./ml, 941.20 ± 19.91 µM Trolox eq./ml, 78.37 µg/g, respectively were significantly higher (p < 0.05) when compared with the extracting medium. The L*, a* and b* values of CRE were 18.65 ± 0.037, 19.42 ± 0.21, 27.947 ± 0.65 and were significantly higher than extracting medium. The CRE could be used as a natural source of ß-carotene and natural colorant for food applications.
ABSTRACT
The concentration of pasteurized buffalo skim milk (PBSM) employing ultrafiltration (UF) alters the chemical composition of ultrafiltered retentate that adversely affect its proteins and salts equilibrium. Effect of stabilizing salts addition in concentrated milks or retentates was majorly dedicated to their thermal stability only. Therefore, this study was aimed to investigate the effect of disodium phosphate (DSP) addition and homogenization of 2.40 × UF retentate (0.60 protein to total solids ratio) on its ζ-potential, particle size, heat stability, turbidity, pH, viscosity and crossover temperature of storage (G') and loss (Gâ³) modulus. Concentration of PBSM in UF process, significantly (P < 0.05) increased its percent TS, protein, fat and ash contents, but markedly decreased its lactose content. DSP addition significantly increased (P < 0.05) the ζ-potential, pH, viscosity and particle size in majority of the homogenized and non-homogenized retentates. Homogenized retentates containing 2.5 and 5% DSP exhibited Newtonian and Power law flow behaviour. However, rheological behaviour of non-homogenized retentates containing zero (control), 1 and 4% DSP was best explained by Bingham model. Further, non-homogenized retentates with 0.5, 2, 3, 5% DSP exhibited Newtonian flow, but retentates containing 6 and 7% DSP was best explained by Power law. The correlation among different attributes of DSP added non-homogenized and homogenized samples were also studied. Particle size and turbidity (r = + 0.999, P < 0.05) as well as ζ-potential and crossover temperature of G' and Gâ³ (r = + 0.999, P < 0.05) showed positive correlation in 4% DSP added non-homogenized retentate.
ABSTRACT
The demand of milk protein concentrate (MPC) powders is continuously increasing as high protein dairy ingredients. Presence of higher calcium and casein contents; heating, ultrafiltration (UF), diafiltration (DF) and spray drying of buffalo skim milk induces undesirable changes in milk proteins that causes problem of poor solubility in MPC powders. Therefore, this investigation was aimed to study the effect of change in pH (6.8-native, 7.0-neutral), heat treatments (74 ± 1 °C/15 s, 80 ± 1 °C/5 min, 85 ± 1 °C/5 min, 90 ± 1 °C/5 min) and DF on physicochemical, functional, reconstitution and rheological properties of medium protein buffalo milk protein concentrate (MP-BMPC) powder. Based on maximum ζ-potential and heat stability, UF retentate was selected, diafiltered and spray dried to obtain MP-BMPC powder. Despite having higher protein content, MP-BMPC powder exhibited markedly better functional (solubility, wettability, viscosity and emulsion stability) properties than buffalo milk protein concentrate 60. The interstitial air content, occluded air content, loose bulk density, packed bulk density, particle density and porosity values of MP-BMPC powder were 145.97 and 112.92 mL 100 g-1 of powder, 0.21 g mL-1, 0.30 g mL-1, 0.55 g mL-1 and 65.09%. Further, its specific surface area; particle size distribution (d10, d50, d90); Sauter (D32) and DeBroukere (D43) mean values were 97.93 m2 kg-1; 34.32, 104.42, 218.58 µm; 61.27 µm and 117.99 µm. The storage modulus (G') and loss modulus (Gâ³) crossover temperature of UF and DF retentates were ~ 57.16 °C and 55.10 °C, respectively. Rheological behaviour of UF, DF retentates and MP-BMPC solution were best explained by Herschel-Bulkley model. Fourier-transform infrared spectroscopy best described amide I, II and III regions in 1700-1400 cm-1 and 1350-1200 cm-1 wavenumber range.
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Ghee forms an important component of the diet of human beings due to its rich flavor and high nutritive value. This high priced fat is prone to adulteration with cheaper fats. ATR-FTIR spectroscopy coupled with chemometrics was applied for determining the presence of goat body fat in ghee (@1, 3, 5, 10, 15 and 20% level in the laboratory made/spiked samples). The spectra of pure (ghee and goat body fat) and spiked samples were taken in the wavenumber range of 4000-500 cm-1. Separated clusters of pure ghee and spiked samples were obtained on applying principal component analysis at 5% level of significance in the selected wavenumber range (1786-1680, 1490-919 and 1260-1040 cm-1). SIMCA was applied for classification of samples and pure ghee showed 100% classification efficiency. The value of R2 was found to be >0.99 for calibration and validation sets using partial least square method at all the selected wavenumber range which indicate that the model was well developed. The study revealed that the spiked samples of goat body fat could be detected even at 1% level in ghee.
ABSTRACT
The objective of the present study was to develop a stable flaxseed oil emulsion for the delivery of omega-3 (ω-3) fatty acids through food fortification. Oil-in-water emulsions containing 12.5 % flaxseed oil, 10 % lactose and whey protein concentrate (WPC)-80 ranging from 5 to 12.5 % were prepared at 1,500, 3,000 and 4,500 psi homogenization pressure. Flaxseed oil emulsions were studied for its physical stability, oxidative stability (peroxide value), particle size distribution, zeta (ζ)-potential and rheological properties. Emulsions homogenized at 1,500 and 4,500 psi pressure showed oil separation and curdling of WPC, respectively, during preparation or storage. All the combinations of emulsions (homogenized at 3,000 psi) were physically stable for 28 days at 4-7 ºC temperature and did not show separation of phases. Emulsion with 7.5 % WPC showed the narrowest particle size distribution (190 to 615 nm) and maximum zeta (ζ)-potential (-33.5 mV). There was a slight increase in peroxide value (~20.98 %) of all the emulsions (except 5 % WPC emulsion), as compared to that of free flaxseed oil (~44.26 %) after 4 weeks of storage. Emulsions showed flow behavior index (n) in the range of 0.206 to 0.591, indicating higher shear thinning behavior, which is a characteristic of food emulsions. Results indicated that the most stable emulsion of flaxseed oil (12.5 %) can be formulated with 7.5 % WPC-80 and 10 % lactose (filler), homogenized at 3,000 psi pressure. The formulated emulsion can be used as potential omega-3 (ω-3) fatty acids delivery system in developing functional foods such as pastry, ice-creams, curd, milk, yogurt, cakes, etc.
ABSTRACT
Thiocyanate is one of the three components of lactoperoxidase system for preserving the milk under the conditions where chilling facilities do not exist due to economic and technical reasons. In 1988, International Dairy Federation (IDF) has recommended the levels and the code of practices for the application of LP-system. As the excessive use of thiocyanates beyond the recommended limits can lead to human health hazards, its rapid detection in milk at the reception dock of dairy, where milk is tested for quality before its acceptance or rejection, is necessary. For simple and rapid detection of added thiocyanates in milk, two types of qualitative tests have been developed in this study, of which one is applicable directly to milk and the other is applicable to TCA filtrate of milk. The results of the test reveal that as low as 0.6 ppm of added thiocyanate (equivalent to 1 ppm of potassium thiocyanate) can be detected. The orange to orange-red color obtained in the test solution can be considered the basis of rejecting milk assuming that it has added levels of thiocyanate.
ABSTRACT
The present investigation was undertaken to use whey for the production of nutritious whey-soya milk dahi. Neutralized whey (pH 7.0 ± 0.2) was used to extract soya milk from cleaned, dried, dehulled and steamed soybean in soya milk extraction plant. Whey-soya milk was extracted having whey to soybean ratio of 5:1. It was further diluted with whey to get different whey to soybean ratios i.e. 9:1, 8:1, 7:1 and 6:1. To set dahi, skim milk powder was added to whey-soya milk so as to make the final total solid concentration of 16, 18 and 20 %. Dahi was prepared by inoculating different whey-soya milk ratios with vita brand of dahi. Results revealed that whey-soya milk having whey to soybean ratio of 5:1 and 20 % total solids was found to be most appropriate for preparation of dahi on the basis of time taken for firmly setting of dahi (5 h) and sensory evaluation scores (7.7 on 9 point hedonic scale). Whey-soya milk dahi was found to be at par with milk dahi (control) based on the sensory scores. Whey-soya milk dahi and milk dahi samples were stored at refrigeration temperature and evaluated for changes in physico-chemical (fat, protein, total sugar, total solids, pH, viscosity and syneresis) and sensory (color and appearance, flavor, consistency and overall acceptability) properties at an interval of 2 days up to 10 days. The results of the study revealed that whey-soya milk dahi could be stored up to 10 days with good sensory quality.
ABSTRACT
The present study was carried out to study the effect of incorporation of fibre rich pumpkin powder and guar gum on the farinographic characteristics of wheat flour. The flour and pumpkin powder were assessed for proximate composition, total dietary fibre, minerals and ß-carotene. Pumpkin powder contained appreciable amount of fibre, minerals and ß-carotene. The effects of incorporation of different levels of pumpkin powder and guar gum along with pumpkin powder on farinographic characteristics were studied. Dough development time, dough stability, time to break down and farinograph quality number increased whereas mixing tolerance index decreased with incorporation of pumpkin powder (> 5 %) and guar gum (1.0 and 1.5 %) along with pumpkin powder in the flour. Resistance to extension as well as extensibility of dough prepared increased significantly by adding pumpkin powder (5-15 %) whereas increase in resistance to extension only was noticed with inclusion of guar gum (0.5-1.5 %) to flour containing 5 % pumpkin powder. Results indicated that pumpkin can be processed to powder that can be utilized with guar gum for value addition.
ABSTRACT
Flaxseed is emerging as an important functional food ingredient because of its rich contents of α-linolenic acid (ALA, omega-3 fatty acid), lignans, and fiber. Flaxseed oil, fibers and flax lignans have potential health benefits such as in reduction of cardiovascular disease, atherosclerosis, diabetes, cancer, arthritis, osteoporosis, autoimmune and neurological disorders. Flax protein helps in the prevention and treatment of heart disease and in supporting the immune system. As a functional food ingredient, flax or flaxseed oil has been incorporated into baked foods, juices, milk and dairy products, muffins, dry pasta products, macaroni and meat products. The present review focuses on the evidences of the potential health benefits of flaxseed through human and animals' recent studies and commercial use in various food products.
ABSTRACT
The present investigation was conducted with the objective to study the effects of various treatments and storage conditions on ascorbic acid, total phenols, antioxidant activity and polyphenol oxidase activity of mung bean sprouts. The sprouts subjected to various treatments viz., pulsed electric field (PEF) (10,000 V for 10 s), hot water dip (HWD) (50 °C for 2 min), ethanol vapours (1 h) and UV-Irradiation (10 kJm(-2) in laminar flow chamber for 1 h); and then stored at room (25 ± 1 °C) and low (7 ± 1 °C) temperature conditions. The sprouts were analyzed regularly at 24 h interval till end of shelf life. Different treatments given to sprouts resulted in differential effect on various parameters. The ascorbic acid, total phenols and antioxidant activity were highest in ethanol vapours treated sprouts. There was a general decrease in polyphenol oxidase activity by various treatments. During storage ascorbic acid, total phenols and antioxidant activity of sprouts first increased and then decreased significantly, however, for polyphenol oxidase activity a progressive increase with increase in storage period was observed. The trends were similar at room and low temperature storage conditions. Thus, it can be concluded that the ethanol vapours significantly improved the ascorbic acid content, total phenols and antioxidant activity of mung bean sprouts, both at room as well as low temperature conditions of storage.