Search results for: protein concentration

Authors: Zunnu Raen Akhtar

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Cotton Leaf Curl Disease (CLCuD) is from Gemini virus and is transmitted through whiteflies in cotton. Transgenic cotton containing Antisense Coat Protein (ACP) has been found to show better results against CLCuD in cotton. In current research, Antisense Coat Protein was inserted in cotton plants to observe resistance developed in the cotton plants against CLCuD. T1 generation of plants were observed for its expression in plants. Tests were carried out to observe the expression of Antisense Coat Protein using Polymerase Chain Reaction (PCR) technique and by southern blotting. Whiteflies showing positive Cotton Leaf Curl Virus (CLCV) were reared and released in bioassay on ACP expressing cotton plants under laboratory as well as confined semi-field conditions. Results confirmed the expression of AC protein in PCR and southern blotting. Further laboratory results showed that cotton plants expressing AC protein showed rare incidence of CLCuD infection as compared to control. In the confined semi-field, similar results were observed in AC protein expressing cotton as compared to control. These results explicitly show that ACP can help to tackle the CLCuD issue in the future and further studies on biochemical processes involved in these plants and effects of ACP induction on non-target organisms should also be studied for eco-system.

Keywords: cotton, white flies, antisense coat protein, CLCV

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Authors: Win Nee Phong, Tau Chuan Ling, Pau Loke Show

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From an industrial perspective, the exploitation of microalgae for protein source is of great economical and commercial interest due to numerous attractive characteristics. Nonetheless, the release of protein from microalgae is limited by the multiple layers of the rigid thick cell wall that generally contain a large proportion of cellulose. Thus an efficient cell disruption process is required to rupture the cell wall. The conventional downstream processing methods which typically involve several unit operational steps such as disruption, isolation, extraction, concentration and purification are energy-intensive and costly. To reduce the overall cost and establish a feasible technology for the success of the large-scale production, microalgal industry today demands a more cost-effective and eco-friendly technique in downstream processing. One of the main challenges to extract the proteins from microalgae is the presence of rigid cell wall. This study aims to provide some guidance on the selection of the efficient solvent to facilitate the proteins released during the cell disruption process. The effects of solvent types such as methanol, ethanol, 1-propanol and water in rupturing the microalgae cell wall were studied. It is interesting to know that water is the most effective solvent to recover proteins from microalgae and the cost is cheapest among all other solvents.

Keywords: green, microalgae, protein, solvents

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Authors: Alireza Naseri, Susan L. Holdt, Charlotte Jacobsen

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In 2014, the global amount of carrageenan production was 60,000 ton with a value of US$ 626 million. From this number, it can be estimated that the total dried seaweed consumption for this production was at least 300,000 ton/year. The protein content of these types of seaweed is 5 – 25%. If just half of this total amount of protein could be extracted, 18,000 ton/year of a high-value protein product would be obtained. The overall aim of this study was to develop a technology that will ensure further utilization of the seaweed that is used only as raw materials for carrageenan production as single extraction at present. More specifically, proteins should be extracted from the seaweed either before or after extraction of carrageenan with focus on maintaining the quality of carrageenan as a main product. Different mechanical, chemical and enzymatic technologies were evaluated. The optimized process was implemented in lab scale and based on its results; the new experiments were done a pilot and larger scale. In order to calculate the efficiency of the new upstream multi-extraction process, protein content was tested before and after extraction. After this step, the extraction of carrageenan was done and carrageenan content and the effect of extraction on yield were evaluated. The functionality and quality of carrageenan were measured based on rheological parameters. The results showed that by using the new multi-extraction process (submitted patent); it is possible to extract almost 50% of total protein without any negative impact on the carrageenan quality. Moreover, compared to the routine carrageenan extraction process, the new multi-extraction process could increase the yield of carrageenan and the rheological properties such as gel strength in the final carrageenan had a promising improvement. The extracted protein has initially been screened as a plant protein source in typical food applications. Further work will be carried out in order to improve properties such as color, solubility, and taste.

Keywords: carrageenan, extraction, protein, seaweed

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Authors: Valentin Nicorescu, Nicoleta C. Predescu, Camelia Papuc, Iuliana Gajaila, Carmen D. Petcu

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The oxidation of the fresh muscle oxymyoglobin (bright red colour) to metmyoglobin (brown colour) leads to discoloration of red meats. After slaughter, enzymatic systems involved in metmyoglobin reduction are continually depleted as time post-mortem progresses, thus the meat colour is affected. Phenolic compounds are able to scavenge reactive species involved in oxymyoglobin oxidation and to reduce metmyoglobin to oxymyoglobin. The aim of this study was to investigate the effect of polyphenols extracted from hawthorn fruits on the stability of oxymyoglobin and myofibrillar proteins in ground pork subject to refrigeration for 6 days. Hawthorn polyphenols (HP) were added in ground pork in 100, 200 and 300 ppm concentrations. Oxymyoglobin and metmyoglobin were evaluated spectrophotometrically at every 2 days and electrophoretic pattern of myofibrillar proteins was investigated at days 0 and 6 by Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis (SDS-PAGE). For all meat samples, oxymyoglobin concentration significantly decreased during the first 4 days of refrigeration. After 6 days, the significant decrease of oxymyoglobin concentration continued only in the negative control samples. In samples treated with HP and butylated hydroxylanisole (BHA - positive control), oxymyoglobin concentration increased after 6 days of refrigeration, the highest levels complying with the following order: 100 ppm HP > 200 ppm HP > 300 ppm HP > 100 ppm BHA. The increase in metmyoglobin was coincidental with the decrease in oxymyoglobin; metmyoglobin concentration progressively increased during the first 4 days of refrigeration in all meat samples. After 6 days, in meat samples treated with HP and BHA, lower metmyoglobin concentrations were found (compared to day 4), respecting the following order: 100 ppm HP < 200 ppm HP < 300 ppm HP < 100 ppm BHA. These results showed that hawthorn polyphenols and BHA reduced metmyoglobin (MbFe3+) to oxymyoglobin (MbFe2+), and the strongest reducing character was recorded for 100 ppm HP. After 6 days of refrigeration, electrophoretic pattern of myofibrillar proteins showed minor changes compared to day 0, indicating that HP prevent protein degradation as well as synthetic antioxidant BHA. Also, HP did not induce cross-links in the myofibrillar proteins, to form protein aggregates, and no risk of reducing their ability to retain water was identified. The pattern of oxymyoglobin and metmyoglobin concentrations determined in this study showed that hawthorn polyphenols are able to reduce metmyoglobin to oxymyoglobin and to delay oxymyoglobin oxidation, especially when they are added to ground meat in concentration of 100 ppm. This work was carried out through Partnerships in priority areas Program – PN II, implemented with the support of MEN – UEFISCDI (Romania), project nr. 149/2014.

Keywords: Hawthorn polyphenols, metmyoglobin, oxymyoglobin, proteins stability

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Authors: Nagia Ali, R. S. R. El-Mohamedy

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The purpose of this paper is to irradiate the dyes biologically through the use of Moreinga oleifera. The study confirms the potential use of Moringa oleifera in decolourization of dyes and thus opens up a scope for future analysis pertaining to its performance in treatment of textile effluent. In this paper, the ability of natural products in removing dyes was tested using two reactive dyes and one acid dye. After a preliminary screening for dye removal capacity, a vegetal protein extract derived from Moeringa oleifera seed was fully studied. The influences of several parameters such as pH, temperature or initial dye concentration were tested and the behavior of coagulants was compared. It was found that dye removal decreased as pH increased. Temperature did not seem to have a considerable effect, while initial dye concentration appeared to be a very important variable.

Keywords: Moreinga oleifera, decolourization, waste water, reactive dyes, acid dyes

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Authors: Lukman Ahmad Jamil, Heather M. Wallace

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Introduction: Numerous epidemiological studies have shown a positive as well as negative association and no association in some cases between chronic use of calcium channel blockers and the increased risk of developing cancer. However, these associations were enmeshed with controversies in the absence of laboratory based studies to back up those claims. Aim: The aim of this study was to determine in mechanistic terms the association between the long-term administration of nifedipine and diltiazem and increased risk of developing cancer using the human embryonic kidney (HEK293) cell line. Methods: Cell counting using the Trypan blue dye exclusion and 3-4, 5-Dimethylthiazol-2-yl-2, 5-diphenyl-tetrazolium bromide (MTT) assays were used to investigate the effect of nifedipine and diltiazem on the growth pattern of HEK293 cells. Protein assay using modified Lowry method and analysis of intracellular polyamines concentration using Liquid Chromatography – Tandem Mass Spectrometry (LC-MS) were performed to ascertain the mechanism through which chronic use of nifedipine increases the risk of developing cancer. Results: Both nifedipine and diltiazem significantly increased the proliferation of HEK293 cells dose and time dependently. This proliferative effect after 24, 48 and 72-hour incubation period was observed at 0.78, 1.56 and 25 µM for nifedipine and 0.39, 1.56 and 25 µM for diltiazem, respectively. The increased proliferation of the cells was found to be statistically significantly (p<0.05). Furthermore, the increased proliferation of the cells induced by nifedipine was associated with the increase in the protein content and elevated intracellular polyamines concentration level. Conclusion: The chronic use of nifedipine is associated with increased proliferation of cells with concomitant elevation of polyamines concentration and elevated polyamine levels have been implicated in many malignant transformations and hence, these provide a possible explanation on the link between long term use of nifedipine and development of some human cancers. Further studies are needed to evaluate the cause of this association.

Keywords: cancer, nifedipine, polyamine, proliferation

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Authors: Ji-Hyon Kil, Kew-Cheol Shim, Kyoung-Ae Park, Kyoungho Kim

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Ambrosia trifida L. is designated as invasive alien species by the Act on the Conservation and Use of Biodiversity by the Ministry of Environment, Korea. The purpose of present paper was to investigate the inhibitory effects of aqueous extracts of A.trifida on the development of root hairs of Triticum aestivum L., and Allium tuberosum Rottler ex Spreng and the electrophoretic protein patterns of their radicles. The development of root hairs was inhibited by increasing of aqueous extract concentrations. Through SDS-PAGE, the electrophoretic protein bands of extracted proteins from their radicles were appeared in controls, but protein bands of specific molecular weight disappeared or weakened in treatments. In conclusion, inhibitory effects of A. trifida made two receptor species changed morphologically, and at the molecular level in early growth stage.

Keywords: Ambrosia trifida L., invasive alien species, inhibitory effect, root hair, electrophoretic protein, radicle

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Authors: Sivapan Choo-In

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The main objective of this study was investigate particulate matter concentration on main and secondary roadside in urban area. And studied on the concentration of some heavy metal including lead (Pb), zinc (Zn), copper (Cu) and cadmium (Cd) in particulate matter in Bangkok area. The averaged particle concentration for main roadside are higher than secondary roadside. The particulate matter less than 10 micron concentration contribute the majority of the Total Suspended Particulate for main road and zinc concentration were higher than copper and lead for both site.

Keywords: air pollution, air quality, polution, monitoring

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Authors: Tatiana R. Simonyan, Elena A. Protasova, Anastasia V. Mamontova, Eugene G. Maksimov, Konstantin A. Lukyanov, Alexey M. Bogdanov

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Here, we describe two variants of the calcium indicators based on the GCaMP sensitive core and BrUSLEE fluorescent protein (GCaMP-BrUSLEE and GCaMP-BrUSLEE-145). In contrast to the conventional GCaMP6-family indicators, these fluorophores are characterized by the well-marked responsiveness of their fluorescence decay kinetics to external calcium concentration both in vitro and in cellulo. Specifically, we show that the purified GCaMP-BrUSLEE and GCaMP-BrUSLEE-145 exhibit three-component fluorescence decay kinetics, with the amplitude-normalized lifetime component (t3*A3) of GCaMP-BrUSLEE-145 changing four-fold (500-2000 a.u.) in response to a Ca²⁺ concentration shift in the range of 0—350 nM. Time-resolved fluorescence microscopy of live cells displays the two-fold change of the GCaMP-BrUSLEE-145 mean lifetime upon histamine-stimulated calcium release. The aforementioned Ca²⁺-dependence calls considering the GCaMP-BrUSLEE-145 as a prospective Ca²⁺-indicator with the signal read-out in the time domain.

Keywords: calcium imaging, fluorescence lifetime imaging microscopy, fluorescent proteins, genetically encoded indicators

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Authors: Serap Pektas

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Ataxia telangiectasia mutated (ATM) is a serine-threonine kinase, which is the major regulator of the DNA damage response. ATM is activated upon the formation of DNA double-strand breaks (DSBs) in the cells. ATM phosphorylates the proteins involved in apoptotic responses, cell cycle checkpoint control, DNA repair, etc. Tumor protein p53, known as p53 is one of these proteins that phosphorylated by ATM. Phosphorylation of p53 at Ser15 residue leads to p53 stabilization in the cells. Often enzymes activity is affected by hydrogen ion concentration (pH). In order to find the optimal pH range for ATM activity, steady-state kinetic assays were performed at acidic and basic pH ranges. Ser15 phosphorylation of p53 is determined by using ELISA. The results indicated that the phosphorylation rate was better at basic pH range compared with the acidic pH range. This could be due to enzyme stability, or enzyme-substrate interaction is pH dependent.

Keywords: ataxia telangiectasia mutated, DNA double strand breaks, DNA repair, tumor protein p53

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Authors: Sharon N. Nuñal, May Flor S. Muegue, Nizzy Hope N. Cartago, Raymund B. Parcon, Sheina B. Logronio

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The antioxidant and anti-inflammatory potentials of Perna Viridis, Modiolus philippinarum, and Mytella charruana found in the Philippines were assessed. Mussel protein samples were hydrolyzed using trypsin, maturase, alcalase and pepsin at 1% and 2% concentrations and then fractionated through membrane filtration (<10 kDa and <30 kDa). Antioxidant assays showed that pepsin hydrolysate at 2% enzyme concentration exhibited the maximum activities for both 2,2-Diphenyl-1-picrylhydrazyl (DPPH) Radical Scavenging Activity (155-176 µM TE/mg protein) and 2,2-azinobis-(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) radical scavenging (67-68 µM TE/mg protein) assays while trypsin hydrolysate dominated the Ferric Reducing Antioxidant Power (FRAP) for the three mussel species. Lower molecular weight peptide fractions at <10 kDa exhibited better antioxidant activities than the higher molecular weight fractions. The anti-inflammatory activities of M. philippinarum and M. charruana showed comparable protein denaturation inhibition potentials with the highest in P. Viridis samples (98.93%). The 5-Lipoxygenase (5-LOX) inhibitory activities of mussel samples showed no significant difference with inhibition exceeding 70%. P. Viridis demonstrated the highest inhibition against Cyclooxygenase-2 (COX-2) at 56.19%, while the rest showed comparable activities. This study showed that the three mussel species are potential sources of bioactive peptides and lipids with antioxidant and anti-inflammatory properties.

Keywords: anti-inflammatory, antioxidant, bioactive properties, mussel

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Authors: Owonikoko A. D., Odoje O. F.

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Fresh sample of Eryngium alpinum was purchased and processed for leaf protein concentrates with a view to evaluating its nutritional potential, mineral composition, amino acid characteristics and phytochemical constituents. Using standard analytical methods. The proximate composition of the leaf protein concentrates revealed moisture content;(5.35±0.21)g/100g, ash;(11.37±0.43)g/100g, crude protein;(48.17±0.46)g/100g, crude fat;(15.38±0.07)g/100g, crude fibre (3.05±0.46)g/100g, and Nitrogen free extractive; (16.68±0.30) g/100g. The mineral content was: Na;(51.88±0.23) mg/100g, K;(65.40±0.32)mg/100g, Ca; (86.89±0.46)mg/100g, Mg;(49.27±0.42) mg/100g, Zn;(0.62±0.03)mg/100g, Fe (6.65±0.43)mg/100g, Mn;(0.96±0.54)mg/100g, Cd;(0.28±0.04)mg/100g, P; (8.55±0.97)mg/100g, while selenium, lead and mercury were not detected in the sample indicating that the sample is free of causing risk of metal poisoning. The results of phytochemical constituents showed phytate; (18.34±0.36)mg/100g, flavonoid (0.25±0.41)mg/100g. The sample contain both essential and non-essential amino acid, with the highest value of Glutamic acid (12.26) and the lowest value of Tryptophan 1.05. the content of the leaf protein content shows that the sample is fit for dietary consumption and could as well be processed to be used as food additives.

Keywords: mineral composition, phytochemical analysis, leaf protein concentrates, eryngium alpinum

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Authors: Akram Abi, Clarissa Müller, Hans-Joachim Jördening

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Laminaribiose is a beta-1,3-glycoside which is used in the medical field for the treatment of dermatitis and also can be used as a building block for new pharmaceutics. The conventional process of laminaribiose production is the uneconomical process of hydrolysis of laminarin extracted from natural polysaccharides of plant origin. A more economical approach however is attainable by enzymatically synthesis of laminaribiose via a reverse phosphorylase reaction catalyzed by laminaribiose phosphorylase (LP) from Euglena gracilis. Different cultivation methods of Euglena gracilis and the effect on LP production have been investigated. Buffered/unbuffered heterotrophic and mixotrophic cultivations of Euglena gracilis has been carried out. Changes of biomass and LP production, glucose level and pH, cell count and shape has been monitored in the course of time. The results obtained from experiments each in three repetitions, show that in the heterotrophic cultivation of Euglena gracilis not only more biomass is produced compared to mixotrophic cultivation, but also higher specific protein concentration is achieved. Furthermore, the LP activity test showed that the protein extracted from heterotrophically cultured cells has a higher LP activity. It was also observed that the cells develop in a distinctive different shape between these two cultures and have different length to width ratios. Taking the heterotrophic culture as the more efficient cultivation method in LP production, another comparative experiment between buffered and unbuffered heterothrophic culture was carried out that showed the unbuffered culture has advantages over the other one in respect of both LP production and resulting activity. A hetrotrophic cultivation of Euglena gracilis in a 5L bioreactor with controlled operating conditions showed a distinctive improvement of all the aspects of culture compared to the shaking flask cultivations. Biomass production was improved from 5 to more than 8 g/l (dry weight) which resulted in a specific protein concentration of 45 g/l in the heterotrophic cultivation in the bioreactor. In further attempts to improve LP production, different purification methods were tested and each method was checks through an activity assay. A laminaribiose yield of 35% was achieved which was by far the highest amount amongst different methods tested.

Keywords: euglena gracilis, heterotrophic culture, laminaribiose production, mixotrophic culture

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Authors: Ehab El-Dabaa, Omnia Ali, Mohamed Abd El-Hady, Ahmed Osman

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Recombinant human interferon alpha 2 (rhIFN-α2) is FDA approved for treatment of some viral and malignant diseases. Approved pegylated rhIFN-α2 drugs have highly improved pharmacokinetics, pharmacodynamics and therapeutic efficiency compared to native protein. In this work, we studied the pegylation of purified properly refolded rhIFN-α2b using linear 20kDa PEG-NHS (polyethylene glycol- N-hydroxysuccinimidyl ester) to prepare pegylated rhIFN-α2b with high stability and activity. The effect of different parameters like rhIFN-α2b final concentration, pH, rhIFN-α2b/PEG molar ratios and reaction time on the efficiency of pegylation (high percentage of monopegylated rhIFN-α2b) have been studied in small scale (100µl) pegylation reaction trials. Study of the percentages of different components of these reactions (mono, di, polypegylated rhIFN-α2b and unpegylated rhIFN-α2b) indicated that 2h is optimum time to complete the reaction. The pegylation efficiency increased at pH 8 (57.9%) by reducing the protein concentration to 1mg/ml and reducing the rhIFN-α2b/PEG ratio to 1:2. Using larger scale pegylation reaction (65% pegylation efficiency), ion exchange chromatography method has been optimized to prepare and purify the monopegylated rhIFN-α2b with high purity (96%). The prepared monopegylated rhIFN-α2b had apparent Mwt of approximately 65 kDa and high in vitro antiviral activity (2.1x10⁷ ± 0.8 x10⁷ IU/mg). Although it retained approximately 8.4 % of the antiviral activity of the unpegylated rhIFN-α2b, its activity is high compared to other pegylated rhIFN-α2 developed by using similar approach or higher molecular weight branched PEG.

Keywords: antiviral activity, rhIFN-α2b, pegylation, pegylation efficiency

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Authors: Illahi Bakhsh Marghazani, Nasrullah, Masood Ul Haq Kakar, Abdul Hameed Baloch, Ahmad Nawaz Khoso, Behram Chacher

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Protein sources are the major part of ration fed to dairy buffaloes in Pakistan however, the limited availability and lack of judicious use of protein resources are further aggravating the conditions to enhance milk and meat production. In order to gain maximum production from limited protein source availability, it is necessary to balance feed for rumen degradable and rumen undegradable protein fractions. This study planned to know the rumen degradable and rumen undegradable fractions in all vegetable protein sources with (formaldehyde and heat treatment) and without treatments. Samples of soybean meal, corn gluten meal 60%, maize gluten feed, guar meal, sunflower meal, rapeseed meal, rapeseed cake, canola meal, cottonseed cake, cottonseed meal, coconut cake, coconut meal, palm kernel cake, almond cake and sesame cake were collected from ten different geographical locations of Pakistan. These samples were also subjected to formaldehyde (1% /100g CP of test feed) and heat treatments (1 hr at 15 lb psi/100 g CP of test feed). In situ technique was used to know the ruminal degradability characteristics. Data obtained were fitted to Orskove equation. Results showed that both treatments significantly (P < 0.05) decreased ruminal degradability in all vegetable protein sources than untreated vegetable protein sources, however, of both treatments, heat treatment was more effective than formaldehyde treatment in decreasing ruminal degradability in most of the studied vegetable protein sources.

Keywords: formaldehyde and heat treatments, in situ technique, rumen degradable and rumen undegradable fractions, vegetable protein sources

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Authors: Roike Iwan Montolalu, Henny Adeleida Dien, Feny Mentang, Kristhina P. Rahael, Tomy Moga, Ayub Meko, Siegfried Berhimpon

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In the last 20 years, packaging materials derived from petrochemicals polymers were widely used as packaging materials. This due to various advantages such as flexible, strong, transparent, and the price is relatively cheap. However, the plastic polymer also has various disadvantages, such as the transmission monomer contamination into the material to be packed, and waste is non-biodegradable. Edible film (EF) is an up to date materials, generated after the biodegradable packaging materials. The advantages of the EF materials, is the materials can be eat together with food, and the materials can be applied as a coating materials for a widely kind of foods especially snack foods. The aims of this research are to produce and to analyze the characteristics of smoked EF made from carrageenan, myofibril and collagen of Black Marlin (Makaira indica) industrial waste. Smoked EF made with an addition of 0.8 % smoke liquid. Three biopolymers i.e. carrageenan, myofibril, and collagen were used as treatments, and homogenate for 1 hours at speed of 1500 rpm. The analysis carried out on the pH and physical properties i.e. thickness, solubility, tensile strength, % elongation, and water vapor transmission rate (WVTR), as well as on the sensory characteristics of texture i.e. wateriness, firmness, elasticity, hardness, and juiciness of the coated products. The result shown that the higher the concentration the higher the thickness of EF, where as for myofibril proteins appeared higher than carrageenan and collagen. Both of collagen and myofibril shown that concentration of 6% was most soluble, while for carrageenan were in concentration of 2 to 2.5%. For tensile strength, carrageenan was significantly higher than myofibril and collagen; while for elongation, collagen film more elastic than carragenan and myofibril protein. Water vapor transmission rate, shown that myofibril protein film lower than carrageenan and collagen film. From sensory assessment of texture, carrageenan has a high elasticity and juiciness, while collagen and myofibril have a high in firmness and hardness.

Keywords: edible film, collagen, myofibril, carrageenan

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Authors: M. N. Hanina, M. Hairul Shahril, I. Ismatul Nurul Asyikin, A. K. Abdul Jalil, M. R. Salina, M. R. Maryam, M. Rosfarizan

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The extracellular proteins secreted by bacteria may be increased in stressful surroundings, such as in the presence of antibiotics. It appears that many antibiotics, when used at low concentrations, have in common the ability to activate or repress gene transcription, which is distinct from their inhibitory effect. There have been comparatively few studies on the potential of antibiotics as a specific chemical signal that can trigger a variety of biological functions. Therefore, this study was carried out to determine the effect of Streptomycin Sulfate in regulating extracellular proteins secreted by Bacillus subtilis ATCC21332. Results of Microdilution assay showed that the Minimum Inhibition Concentration (MIC) of Streptomycin Sulfate on B. subtilis ATCC21332 was 2.5 mg/ml. The bacteria cells were then exposed to Streptomycin Sulfate at concentration of 0.01 MIC before being further incubated for 48h to 72 h. The extracellular proteins secreted were then isolated and analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Proteins profile revealed that three additional bands with approximate sizes of 30 kDa, 22 kDa and 23 kDa were appeared for the treated bacteria with Streptomycin Sulfate. Thus, B. subtilis ATCC21332 in stressful condition with the presence of Streptomycin Sulfate at low concentration could induce the extracellular proteins secretion.

Keywords: Bacillus subtilis ATCC21332, streptomycin sulfate, extracellular proteins, antibiotics

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Authors: F. Yelles-Allam, A. A. Nouani

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In Algeria, whey is discarded without any treatment and this causes not only pollution problem, but also a loss in nutritive components of milk. In this paper, characterization of EDAM whey, which is resulted from pasteurised mixture of cow’s milk and skim milk, and recovery of whey protein by ultrafiltration / diafiltration, was studied. The physical-chemical analysis of whey has emphasized on its pollutant and nutritive characteristics. In fact, its DBO5 and DCO are 49.33, and 127.71 gr of O2/l of whey respectively. It contains: fat (1,90±0,1 gr/l), lactose (47.32±1,57 gr/l), proteins (8.04±0,2 gr/l) and ashes (5,20±0,15 gr/l), calcium (0,48±0,04 gr/l), Na (1.104gr/l), K (1.014 gr/l), Mg (0.118 gr/l) and P (0.482 gr/l). Ultrafiltration was carried out in a polyetersulfone membrane with a cut-off of 10K. Its hydraulic intrinsic resistance and permeability are respectively: 2.041.1012 m-1 and 176,32 l/h.m2 at PTM of 1 bar. The retentate obtained at FC6, contains 16,33g/l of proteins and 70,25 g/l of dry matter. The retention rate of protein is 97, 7% and the decrease in DBO5 and DCO are at 18.875 g /l and 42.818 g/l respectively. Diafiltration performed on protein concentrates allowed the complete removal of lactose and minerals. The ultrafiltration of the whey before the disposal is an alternative for Algéria dairy industry.

Keywords: diafiltration, DBO, DCO, protein, ultrafiltration, whey

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Authors: Andi Aliah Hidayani, Asmi Citra Malina A. R. Tassakka, Andi Parenrengi

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Vanname Shrimp is one of the high yielding varieties that are more resistant to virus attacks. However, now this shrimp more death due to virus attack such as white spot disease caused by white spot syndrome virus (WSSV). Various efforts have done to prevent the disease, like immunostimulatory, probiotics, and vaccine. White spot syndrome virus (WSSV) envelope protein VP19 gene is important because of its involvement in the system infection of shrimp. This study aimed to isolate and characterize an envelope protein VP19 – encoding gene of WSSV using WSSV infected Vanname Shrimp sample from some areas in South Sulawesi (Pangkep, Barru and Pinrang). The genomic of DNA were isolated from shrimp muscle using DTAB-CTAB method. Isolation of gene encoding envelope protein VP19 WSSV ws successfully performed with the results of the length of DNA fragment was 387 bp. The results of homology analysis using BLASTn homology suggested that these isolates genes from Barru, Pangkep and Pinrang have closest relationship with isolates from Mexican.

Keywords: vanname, shrimp, WSSV, viral protein 19

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Authors: Rahamdad Khan, Ijaz Ahmad Khan

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In modern agriculture the herbicides application are considered the most effective and fast in action against all types of weeds. But it’s a fact that the herbicide applicator cannot totally secure the crop plants from the possible herbicide injuries that further leads to several destructive changes in plant biochemistry. For the purpose pots studies were undertaken to test the tolerance order of chickpea against pre- emergence herbicides (Stomp 330 EC- Dual Gold 960 EC) and post- emergence herbicides (Topik 15 WP- Puma Super 75 EW- Isoproturon 500 EW) during 2012-13 and 2013-14. The experimental design was CRD with three replications. Plant height, number of branches plant-1, number of seeds plant-1, nodulation, seed protein contents and other growth related parameters in chickpea were examined during the investigations. The results indicate that all the enquire herbicides gave a significant variation to all recorded parameter of chick pea except nodule fresh and dray weight. Moreover the toxic effect of pre-emergence herbicide on chickpea was found higher as compared to post-emergence herbicides. Minimum chickpea plant height (50.50 cm), number of nodule plant-1 (17.83) and lowest seed protein (14.13 %) was recorded in Stomp 330 EC. Similarly the outmost seeds plant-1 (29.66) and number of nodule plant-1 (21) were found for Puma Super 75 EW. The results further showed that the highest seed protein content (21.75 and 21.15 %) was recorded for control/ untreated and Puma Super 75EW. Taking under concentration the possible negative impact of the herbicides the chemical application must be minimized up to certain extent at which the crop is mostly secure. However chemical weed control has many advantages so we should train our farmer regarding the proper use of agro chemical to minimize the loses in crops while using herbicides.

Keywords: chickpea, herbicides, protein, stomp 330 EC, weed

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Authors: Aylin W Sahin, Alice Jaeger, Laura Nyhan, Gregory Belt, Steffen Münch, Elke K. Arendt

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Our current food systems cause an increase in malnutrition resulting in more people being overweight or obese in the Western World. Additionally, our natural resources are under enormous pressure and the greenhouse gas emission increases yearly with a significant contribution to climate change. Hence, transforming our food systems is of highest priority. Plant-based food products have a lower environmental impact compared to their animal-based counterpart, representing a more sustainable protein source. However, most plant-based protein ingredients, such as soy and pea, are lacking indispensable amino acids and extremely limited in their functionality and, thus, in their food application potential. They are known to have a low solubility in water and change their properties during processing. The low solubility displays the biggest challenge in the development of milk alternatives leading to inferior protein content and protein quality in dairy alternatives on the market. Moreover, plant-based protein ingredients often possess an off-flavour, which makes them less attractive to consumers. EverPro, a plant-protein isolate originated from Brewer’s Spent Grain, the most abundant by-product in the brewing industry, represents the missing piece in the plant protein portfolio. With a protein content of >85%, it is of high nutritional value, including all indispensable amino acids which allows closing the protein quality gap of plant proteins. Moreover, it possesses high techno-functional properties. It is fully soluble in water (101.7 ± 2.9%), has a high fat absorption capacity (182.4 ± 1.9%), and a foaming capacity which is superior to soy protein or pea protein. This makes EverPro suitable for a vast range of food applications. Furthermore, it does not cause changes in viscosity during heating and cooling of dispersions, such as beverages. Besides its outstanding nutritional and functional characteristics, the production of EverPro has a much lower environmental impact compared to dairy or other plant protein ingredients. Life cycle assessment analysis showed that EverPro has the lowest impact on global warming compared to soy protein isolate, pea protein isolate, whey protein isolate, and egg white powder. It also contributes significantly less to freshwater eutrophication, marine eutrophication and land use compared the protein sources mentioned above. EverPro is the prime example of sustainable ingredients, and the type of plant protein the food industry was waiting for: nutritious, multi-functional, and environmentally friendly.

Keywords: plant-based protein, upcycled, brewers' spent grain, low environmental impact, highly functional ingredient

Procedia PDF Downloads 47

Authors: Alka Yadav, Mayank Jain, Rajan Saxena, Niraj Kumari, Narendra Krishnani, Ashok Kumar

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Purpose: To study the mismatch repair (MMR) protein expression and its clinicopathological correlation in colorectal cancer patients in North India. Methods: A prospective study was conducted on histologically proven 52 (38 males and 14 females) patients with adenocarcinoma of colorectum. MMR protein loss was determined by using immunohistochemistry for MLH1, MSH2, PMS2 and MSH6. Results: 52 patients (38 males and 14 females) underwent resection for colorectal cancer with the median age of 52 years (16-81 years). 35% of the patients (n=18) were younger than 50 years of the age. 3 patients had associated history of malignancy in the family. 29 (56%) patients had right colon cancer, 9 (17%) left colon cancer and 14 (27%) rectal cancer. 2 patients each had synchronous and metachronous cancer. Histology revealed well-differentiated tumour in 16, moderately differentiated in 10 and poorly differentiated tumour in 26 patients. MMR protein loss was seen in 15 (29%) patients. Seven (46%) of these patients were less than 50 years of age. Combined loss of MSH2 and MSH6 was seen most commonly and it was found in 6 patients. 12 (80%) patients with MMR protein loss had tumour located proximal to the splenic flexure compared to 3 (20%) located distal to the splenic flexure. There was no difference in MMR protein loss based on patients' age, gender, degree of tumour differentiation, stage of the disease and tumour histological characteristics. Conclusions: This study revealed that there was less than 30% MMR protein loss in colorectal cancer patients. The loss was most commonly seen in right sided colon cancer than left. A larger study is further required to validate these findings.

Keywords: colorectal cancer, mismatch repair protein, immunohitochemistry, clinicopathological correlation

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Authors: Hossein Rassi, Marjan Moradi Fard, Samaneh Niko

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Human papillomavirus type 16 and 18 are closely associated with the development of human cervical carcinoma, which is one of the most common causes of cancer death in women worldwide. At present, HPV type 18 accounts for about 34 % of all HPV infections in Iran and the most promising vaccine against HPV infection is based on the L1 major capsid protein. The L1 protein of HPV18 has the capacity to self-assemble into capsomers or virus-like particles (VLPs) that are non-infectious, highly immunogenic and allowing their use in vaccine production. The methylotrophic yeast Pichia pastoris is an efficient and inexpensive expression system used to produce high levels of heterologous proteins. In this study we expressed HPV18 L1 VLPs in P. pastoris. The gene encoding the major capsid protein L1 of the high-risk HPV type 18 was isolated from Iranian patient by PCR and inserted into pTG19-T vector to obtain the recombinant expression vector pTG19-HPV18-L1. Then, the pTG19-HPV18-L1 was transformed into E. coli strain DH5α and the recombinant protein HPV18 L1 was expressed under IPTG induction in soluble form. The HPV18 L1 gene was excised from recombinant plasmid with XhoI and EcoRI enzymes and ligated into the yeast expression vector pPICZα linearized with the same enzymes, and transformed into P. pastoris. Induction and expression of HPV18 L1 protein was demonstrated by BMGY/BMMY and RT PCR. The parameters for induced cultivation for strain in P. pastoris KM71 with HPV16L1 were investigated in shaking flask cultures. After induced cultivation BMMY (pH 7.0) medium supplemented with methanol to a final concentration of 1.0% every 24 h at 37 degrees C for 96 h, the recombinant produced 78.6 mg/L of L1 protein. This work offers the possibility for the production of prophylactic vaccine for cervical carcinoma by P. pastoris for HPV-18 L1 gene. The VLP-based HPV vaccines can prevent persistent HPV18 infections and cervical cancer in Iran. The HPV-18 L1 gene was expressed successfully in E.coli, which provides necessary basis for preparing HPV-18 L1 vaccine in human. Also, HPV type 6 L1 proteins expressed in Pichia pastoris will facilitate the HPV vaccine development and structure-function study.

Keywords: Pichia pastoris, L1 virus-like particles, human papillomavirus type 18, biotechnology

Procedia PDF Downloads 381

Authors: Sweta Pandey, Samridhi Dhyani, Susmita Chaudhuri

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Klebsiella pneumoniae bacteria is a global threat to human health due to an increase in multi-drug resistance among strains. The hypervirulent strains of Klebsiella pneumoniae is a major trouble due to their association with life-threatening infections in a healthy population. One of the major virulence factors of hyper virulent strains of Klebsiella pneumoniae is the T6SS (Type six secretary system) which is majorly involved in microbial antagonism and causes interaction with the host eukaryotic cells during infections. T6SS mediates some of the crucial factors for establishing infection by the bacteria, such as cell adherence, invasion, and subsequent in vivo colonisation. The antibacterial activity and the cell invasion property of the T6SS system is a major requirement for the establishment of K. pneumoniae infections within the gut. The T6SS can be an appropriate target for developing therapeutics. The T6SS consists of an inner tube comprising hexamers of Hcp (Haemolysin -regulated protein) protein, and at the top of this tube sits VgrG (Valine glycine repeat protein G); the tip of the machinery consists of PAAR domain containing proteins which act as a delivery system for bacterial effectors. For this study, immune response to recombinant VgrG protein was generated to establish this protein as a potential immunogen for the development of therapeutic leads. The immunogenicity of the selected protein was determined by predicting the B cell epitopes by the BCEP analysis tool. The gene sequence for multiple domains of VgrG protein (phage_base_V, T6SS_Vgr, DUF2345) was selected and cloned in pMAL vector in E. coli. The construct was subcloned and expressed as a fusion protein of 203 residue protein with mannose binding protein tag (MBP) to enhance solubility and purification of this protein. The purified recombinant VgrG fusion protein was used for mice immunisation. The antiserum showed reactivity with the recombinant VgrG in ELISA and western blot. The immunised mice were challenged with K. pneumoniae bacteria and showed bacterial clearance in immunised mice. The recombinant VgrG protein can further be used for studying downstream signalling of VgrG protein in mice during infection and for therapeutic MAb development to eradicate K. pneumoniae infections.

Keywords: immune response, Klebsiella pneumoniae, multi-drug resistance, recombinant protein expression, T6SS, VgrG

Procedia PDF Downloads 70

Authors: Vernie C. Convicto, Henry P. Aringa, Wilson I. Barredo

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This study presents a conformational model of the helical structures of globular protein particularly ferritin in the framework of white noise path integral formulation by using Associated Legendre functions, Bessel and convolution of Bessel and trigonometric functions as modulating functions. The model incorporates chirality features of proteins and their helix-turn-helix sequence structural motif.

Keywords: globular protein, modulating function, white noise, winding probability

Procedia PDF Downloads 443

Authors: B. Coşgun, O. Ozturk

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Canola is a specific edible type of rapeseed, developed in the 1970s, which contains about 40 percent oil. This research was carried out to determine the yield and some quality characteristics of some winter canola cultivars during the 2010-2011 vegetation period in Central Anatolia of Turkey. In this research; Oase, Dante, Californium, Excalibur, Elvis, ES Hydromel, Licord, Orkan, Vectra, Nelson, Champlain and NK Petrol winter canola varieties were used as material. The field experiment was set up in a “Randomized Complete Block Design” with three replications on 21 September 2010. In this research; seed yield, oil content, protein content, oil yield and protein yield were examined. As a result of this research; seed yield, oil content, oil yield and protein yield (except protein content) were significant differences between the cultivars. The highest seed yield (6348 kg ha-1) was obtained from the NK Petrol, while the lowest seed yield (3949 kg ha-1) was determined from the Champlain cultivar was obtained. The highest oil content (46.73%) was observed from Oase and the lowest value was obtained from Vectra (41.87%) cultivar. The highest oil yield (2950 kg ha-1) was determined from NK Petrol while the least value (1681 kg ha-1) was determined from Champlain cultivar. The highest protein yield (1539.3 kg ha-1) was obtained from NK Petrol and the lowest protein yield (976.5 kg ha-1) was obtained from Champlain cultivar. The main purpose of the cultivation of oil crops, to increase the yield of oil per unit area. According the result of this research, NK Petrol cultivar which ranks first with regard to both seed yield and oil yield between cultivars as the most suitable winter canola cultivar of local conditions.

Keywords: rapeseed, cultivar, seed yield, crude oil ratio, crude protein ratio, crude oil yield, crude protein yield

Procedia PDF Downloads 243

Authors: Mohamed Ahmed Tony, Fayez Abaza

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The effects of a dietary supplement of rumen-protected choline on feed intake, milk yield, milk composition and some blood metabolites were evaluated in transition dairy cows. Forty multiparous cows were blocked into 20 pairs and then randomly allocated to either one of 2 treatments. The treatments were supplementation either with or without (control) rumen-protected choline. Treatments were applied from 2 weeks before and until 8 weeks after calving. Both groups received the same basal diet as total mixed ration. Additionally, 50 g of a rumen-protected choline supplement (25% rumen protected choline chloride) was added individually in the feed. Individual feed intake, milk yield, and body weight were recorded daily. Milk samples were analyzed weekly for fat, protein, and lactose content. Blood was sampled at week 2 before calving, d 1, d 4, d 7, d 10, week 2, week 3, and week 8 after calving. Glucose, triglycerids, nonesterified fatty acids, and β-hydroxybutyric acid in blood were analysed. The results revealed that choline supplementation increased DM intake from 16.5 to 18.0 kg/d and, hence, net energy intake from 99.2 to 120.5 MJ/d at the intercept of the lactation curve at 1 day in milk. Choline supplementation had no effect on milk yield, milk fat yield, or lactose yield. Milk protein yield was increased from 1.11 to 1.22 kg/d at the intercept of the lactation curve. Choline supplementation was associated with decreased milk fat concentration at the intercept of the lactation curve at 1 day in milking, but the effect of choline on milk fat concentration gradually decreased as lactation progressed. Choline supplementation decreased the concentration of blood triglycerids during the first 4 wk after parturition. Choline supplementation had no effect on energy-corrected milk yield, energy balance, body weight and body condition score. Results from this study suggest that fat metabolism in periparturient dairy cows is improved by choline supplementation during the transition period and this may potentially decrease the risk for metabolic disorders in the periparturient dairy cow.

Keywords: choline, dairy cattle, transition cow, triglycerids

Procedia PDF Downloads 483

Authors: Ali Taheri Mirghaed, Sara Ahani, Ashkan Zargar, Seyyed Morteza Hoseini

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Diseases are the major challenges in intensive aquaculture that cause significant annual losses. Antibiotic-therapy is a common way to control bacterial disease in fish, and oxytetracycline (OTC) is the only oral antibiotic in aquaculture approved FDA. OTC has been found to have negative effects on fish, such as oxidative stress and immune-suppression, thus, it is necessary to mitigate such effects. Medicinal herbs have various benefits on fish, including antioxidant, immunostimulant, and anti-microbial effects. Therefore, we hypothesized if dietary ginger meal (GM) interacts with dietary OTC by monitoring plasma protein fractions in rainbow trout. The study was conducted as a 2 × 2 factorial design, including diets containing 0 and 1% GM and 0 and 1.66 % OTC (corresponding to 100 mg/kg fish biomass per day). After ten days treating the fish (60 g individual weight) with these feeds, blood samples were taken from al treatments (n =3). Plasma was separated by centrifugation, and protein fractions were determined by electrophoresis. The results showed that OTC and GM had interaction effects on total protein (P<0.001), albumin (P<0.001), alpha-1 fraction (P=0.010), alpha-2 fraction (P=0.001), beta-2 fraction (P=0.014), and gamma fraction (P<0.001). Beta-1 fraction was significantly (P=0.030) affected by dietary GM. GM decreased plasma total protein, albumin, and beta-2 but increased beta-1 fraction. OTC significantly decreased total protein (P<0.001), albumin (P=0.001), alpha-2 fraction (P<0.001), beta-2 fraction (P=0.004), and gamma fraction (P<0.001) but had no significant effects on alpha-1 and beta-1 fractions. Dietary GM inhibited/suppressed the effects of dietary OTC on the plasma total protein and protein fractions. In conclusion, adding 1% GM to diet can mitigate the negative effects of dietary OTC on plasma proteins. Thus, GM may boost health of rainbow trout during the period of medication with OTC.

Keywords: ginger, plasma protein electrophoresis, dietary additive, rainbow trout

Procedia PDF Downloads 53

Authors: Dinara Ikramova, Karin A. Hing, Simon C. F. Rawlinson

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Silicate-substituted hydroxyapatite (SiHA) has been shown to enhance bone regeneration in vivo compared with phase pure stoichiometric hydroxyapatite. Evidence suggests that substrate chemistry dependent formation of a permissive protein layer on the surface of synthetic bone graft substitute materials is key for bioactivity and cell attachment. However, little information is available on whether the substrate chemistry may affect cell migration and recruitment. The aim of this study is to investigate whether or not human Mesenchymal Stem Cells (hMSCs) exhibit a chemotactic response to SiHA porous granules and if it can be linked to either the ion exchange or protein sequestering and enrichment on the surface of the material. 150mg of SiHA granules with 80% total porosity and 20% strut porosity were incubated in 1ml of either Serum Free Media (SFM) or 10% Serum Containing Media (SCM) under static cell culture conditions (37°C, 5% CO2) in absence of cells. Protein sequestering and exchange of calcium, phosphate and silicate ions were analysed at 0.5, 1, 2, 4, 8, 16 and 24 hours with n=12 per time point. Migration of hMSCs in the presence of 150mg of SiHA granules was assessed over 24 hours using a modified transwell migration system in either SFM or SCM (n=6) with 30% serum containing media acting as a positive control. At 24 hours protein sequestering and ionic exchange were analysed, and the number of cells was quantified using a high throughput confocal microscope (IN Cell Analyser 6000). In acellular condition, both calcium and phosphate ion concentrations in media showed a decrease at 24 hours which was greater in SFM than in SCM. This suggests possible formation and precipitation of a bone like apatite on the surface of SiHA. Reduction in this activity observed in SCM indicates that the presence of serum proteins is interfering with the ion exchange at the material and media interface. Adsorbed protein levels showed fluctuation over time followed by sharp decrease at 24 hours, suggesting a possible protein rearrangement on the surface of the material. The ion analysis performed on SFM and SCM after 24-hour incubation with cells in the presence of granules showed a greater reduction in phosphate concentration in both SFM and SCM compared to phosphate levels in acellular condition. Silicate concentration in SCM increased from 1.6mM (absence of cells) to 5.1mM (presence of cells). This indicates that the cells are promoting the uptake of phosphate and release of silicate ions. No significant change was seen in levels of adsorbed proteins in the presence and absence of cells. Further analysis is required to determine whether the species of these proteins change over time. The analysis of cell migration after 24-hour incubation showed more cells migrating towards the granules, 12.7% in SFM and 8.3% in SCM, than in positive control, 4.5% in SFM and 3.6% in SCM respectively. These results suggest that SiHA has a chemotactic activity independent of serum proteins. A property which has not previously been demonstrated for a synthetic bone graft material.

Keywords: cell migration, hMSCs, SiHA, transwell migration system

Procedia PDF Downloads 109

Authors: Silvia Gastaldello, Maria Grillo, Luca Tassoni, Claudio Maran, Stefano Balbo

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Antioxidants are nowadays attractive not only for the countless benefits to the human and animal health, but also for the perspective of use as food preservative instead of synthetic chemical molecules. In this study, the radical scavenging activity of six protein extracts from pulse and oleaginous seeds was evaluated. The selected matrices are Pisum sativum (yellow pea from two different origins), Carthamus tinctorius (safflower), Helianthus annuus (sunflower), Lupinus luteus cv Mister (lupin) and Glycine max (soybean), since they are economically interesting for both human and animal nutrition. The seeds were grinded and proteins extracted from 20mg powder with a specific vegetal-extraction kit. Proteins have been quantified through Bradford protocol and scavenging activity was revealed using DPPH assay, based on radical DPPH (2,2-diphenyl-1-picrylhydrazyl) absorbance decrease in the presence of antioxidants molecules. Different concentrations of the protein extract (1, 5, 10, 50, 100, 500 µg/ml) were mixed with DPPH solution (DPPH 0,004% in ethanol 70% v/v). Ascorbic acid was used as a scavenging activity standard reference, at the same six concentrations of protein extracts, while DPPH solution was used as control. Samples and standard were prepared in triplicate and incubated for 30 minutes in dark at room temperature, the absorbance was read at 517nm (ABS30). Average and standard deviation of absorbance values were calculated for each concentration of samples and standard. Statistical analysis using t-students and p-value were performed to assess the statistical significance of the scavenging activity difference between the samples (or standard) and control (ABSctrl). The percentage of antioxidant activity has been calculated using the formula [(ABSctrl-ABS30)/ABSctrl]*100. The obtained results demonstrate that all matrices showed antioxidant activity. Ascorbic acid, used as standard, exhibits a 96% scavenging activity at the concentration of 500 µg/ml. At the same conditions, sunflower, safflower and yellow peas revealed the highest antioxidant performance among the matrices analyzed, with an activity of 74%, 68% and 70% respectively (p < 0.005). Although lupin and soybean exhibit a lower antioxidant activity compared to the other matrices, they showed a percentage of 46 and 36 respectively. All these data suggest the possibility to use undervalued edible matrices as antioxidants source. However, further studies are necessary to investigate a possible synergic effect of several matrices as well as the impact of industrial processes for a large-scale approach.

Keywords: antioxidants, DPPH assay, natural matrices, vegetal proteins

Procedia PDF Downloads 391