Search results for: antioxidant enzyme activity

Authors: Le Thuy Mai, Vu Nguyen Thanh

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β-Glucosidase is an important enzyme for production of ethanol from lignocellulose. With hydrolytic activity on cellooligosaccharides, especially cellobiose, β-glucosidase removes product inhibitory effect on cellulases and forms fermentable sugars. In this study, β-glucosidase encoding gene (BGL1) from traditional starter yeast Saccharomycosis fibuligera BMQ908 was cloned and expressed in Pichia pastoris. BGL1 of S. fibuligera BMQ 908 shared 98% nucleotide homology with the closest GenBank sequence (M22475) but identity in amino-acid sequences of catalytic domains. Recombinant plasmid pPICZαA/BGL1 containing the sequence encoding BGL1 mature protein and α-factor secretion signal was constructed and transformed into methylotrophic yeast P. pastoris by electroporation. The recombinant strain produced single extracellular protein with molecular weight of 120 kDa and cellobiase activity of 60 IU/ml. The optimum pH of the recombinant β-glucosidase was 5.0 and the optimum temperature was 50°C.

Keywords: β-Glucosidase, Pichia pastoris, Saccharomycopsisfibuligera, recombinant enzyme.

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Authors: Md. Z. Alam, Devi R. Asih, Md. N. Salleh

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Immobilization of lipase enzyme produced from palm oil mill effluent (POME) by the activated carbon (AC) among the low cost support materials was optimized. The results indicated that immobilization of 94% was achieved by AC as the most suitable support material. A sequential optimization strategy based on a statistical experimental design, including one-factor-at-a-time (OFAT) method was used to determine the equilibrium time. Three components influencing lipase immobilization were optimized by the response surface methodology (RSM) based on the face-centered central composite design (FCCCD). On the statistical analysis of the results, the optimum enzyme concentration loading, agitation rate and carbon active dosage were found to be 30 U/ml, 300 rpm and 8 g/L respectively, with a maximum immobilization activity of 3732.9 U/g-AC after 2 hrs of immobilization. Analysis of variance (ANOVA) showed a high regression coefficient (R²) of 0.999, which indicated a satisfactory fit of the model with the experimental data. The parameters were statistically significant at p<0.05.

Keywords: Activated carbon, adsorption, immobilization, POME based lipase.

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Authors: M. Cynthya, V. Prabhawathi, D. Mukesh

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Food contamination occurs during post process handling. This leads to spoilage and growth of pathogenic microorganisms in the food, thereby reducing its shelf life or spreading of food borne diseases. Several methods are tried and one of which is use of antimicrobial packaging. Here, papain, a protease enzyme, is covalently immobilized with the help of glutarldehyde on polyurethane and used as a food wrap to protect food from microbial contamination. Covalent immobilization of papain was achieved at a pH of 7.4; temperature of 4°C; glutaraldehyde concentration of 0.5%; incubation time of 24h; and 50mg of papain. The formation of -C=Nobserved in the Fourier transform infrared spectrum confirmed the immobilization of the enzyme on the polymer. Immobilized enzyme retained higher activity than the native free enzyme. The modified polyurethane showed better reduction of Staphylococcus aureus biofilm than bare polymer film (eight folds reduction in live colonies, two times reduction in protein and 6 times reduction in carbohydrates). The efficacy of this was studied by wrapping it over S. aureus contaminated cottage cheese (paneer) and cheese and stored at a temperature of 4°C for 7days. The modified film reduced the bacterial contamination by eight folds when compared to the bare film. FTIR also indicated reduction in lipids, sugars and proteins in the biofilm.

Keywords: Cheese, Papain, polyurethane, Staphylococcus aureus.

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Authors: P. Pinyaphong, S. Phutrakul

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Palm oil could be converted to cocoa butter equivalent by lipase-catalyzed interesterification. The objective of this research was to investigate the structure modification of palm oil to cocoa butter equivalent using Carica papaya lipase –catalyzed interesterification. The study showed that the compositions of cocoa butter equivalent were affected by acyl donor sources, substrate ratio, initial water of enzyme, reaction time, reaction temperature and the amount of enzyme. Among three acyl donors tested (methyl stearate, ethyl stearate and stearic acid), methyl stearate appeared to be the best acyl donor for incorporation to palm oil structure. The best reaction conditions for cocoa butter equivalent production were : substrate ratio (palm oil : methyl stearate, mol/mol) at 1 : 4, water activity of enzyme at 0.11, reaction time at 4 h, reaction temperature at 45 ° C and 18% by weight of the enzyme. The chemical and physical properties of cocoa butter equivalent were 9.75 ± 0.41% free fatty acid, 44.89 ± 0.84 iodine number, 193.19 ± 0.78 sponification value and melting point at 37-39 °C.

Keywords: Carica papaya lipase, cocoa butter equivalent, interesterification, palm oil.

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Authors: N. N. A. K. Shah, R. A. Rahman, R. Shamsuddin, N. M. Adzahan

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Pummelo is known to be the largest of all citrus fruits, with expected ratio of 2:1 (w/v) of producing juice, is an attractive opportunity for Malaysia to expand pummelo-s influence and marketability over the international market of juices. The purpose of this study is to identify and quantify the phenolic compounds in two Malaysian varieties of pummelo fruit juice: Ledang (PO55) and Tambun (PO52). Identifications of polyphenols composition were done using High Performance Liquid Chromatography Diode Array Detection (HPLC-DAD). The phenolic compounds that were found in both varieties were hydroxycinnamic acids and flavonones. This study proved that Tambun variety has the highest antioxidant and phenolic compounds in comparison to Ledang variety. However, considerations have to be made to suit consumer-s taste bud and the amount of enzyme needed to clarify the juice for its marketability.

Keywords: Antioxidant, HPLC, phenolic contents and pummelo fruit juice

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Authors: Wahyu Widowati, Hana Ratnawati, Tjandrawati Mozefis, Dwiyati Pujimulyani, Yelliantty Yelliantty

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Background: Atherosclerosis is the main cause of cardiovascular disease (CVD) with complex and multifactorial process including atherogenic lipoprotein, oxidized low density lipoprotein (LDL), endothelial dysfunction, plaque stability, vascular inflammation, thrombotic and fibrinolytic disorder, exercises and genetic factor Epidemiological studies have shown tea consumption inversely associated with the development and progression of atherosclerosis. The research objectives: to elucidate hypolipidemic, antioxidant effects, as well as ability to improve coronary artery’s histopathologyof black tea extract (BTE) and quercetin in atherosclerotic rats. Methods: The antioxidant activity was determined by using Superoxide Dismutase activity (SOD) of serum and lipid peroxidation product (Malondialdehyde) of plasma and lipid profile including cholesterol total, LDL, triglyceride (TG), High Density Lipoprotein (HDL) of atherosclerotic rats. Inducing atherosclerotic, rats were given cholesterol and cholic acid in feed during ten weeks until rats indicated atherosclerotic symptom with narrowed artery and foamy cells in the artery’s wall. After rats suffered atherosclerotic, the high cholesterol feed and cholic acid were stopped and rats were given BTE 450; 300; 150 mg/kg body weight (BW) daily, quercetin 15; 10; 5 mg/kg BW daily, compared to rats were given vitamin E 60 mg/kg/BW; simvastatin 2.7 mg/kg BW, probucol 30 mg/kg BW daily for 21 days (first treatment) and 42 days (second treatment), negative control (normal feed), positive control (atherosclerotic rats). Results: BTE and quercetin could lower cholesterol total, triglyceride, LDL MDA and increase HDL, SOD were comparable with simvastatin, probucol both for 21 days and 42 days treatment, as well to improve coronary arteries histopathology. Conclusions: BTE andquercetin have hypolipidemic and antioxidant effects, as well as improve coronary arteries histopathology in atherosclerotic rats.

Keywords: Black tea, quercetin, atherosclerosis, antioxidant, hypolipidemic, cardiovascular disease.

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Authors: Marinos Xagoraris, Iliada K. Lappa, Charalambos Kanakis, Dimitra Daferera, Christina Papadopoulou, Georgios Sourounis, Charilaos Giotis, Pavlos Bouchagier, Christos S. Pappas, Petros A. Tarantilis, Efstathia Skotti

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The aim of this work was to recover phenolic compounds from grape skins produced in Greek varieties of the Ionian Islands in order to form the basis of calculations for their further utilization in the context of the circular economy. Isolation and further utilization of phenolic compounds is an important issue in winery by-products. For this purpose, 37 samples were collected, extracted, and analyzed in an attempt to provide the appropriate basis for their sustainable exploitation. Extraction of the bioactive compounds was held using an eco-friendly, non-toxic, and highly effective water-glycerol solvent system. Then, extracts were analyzed using UV-Vis, liquid chromatography-mass spectrometry (LC-MS), FTIR, and Raman spectroscopy. Also, total phenolic content and antioxidant activity were measured. LC-MS chromatography showed qualitative differences between different varieties. Peaks were attributed to monomeric 3-flavanols as well as monomeric, dimeric, and trimeric proanthocyanidins. The FT-IR and Raman spectra agreed with the chromatographic data and contributed to identifying phenolic compounds. Grape skins exhibited high total phenolic content (TPC), and it was proved that during vinification, a large number of polyphenols remained in the pomace. This study confirmed that grape skins from Ionian Islands are a promising source of bioactive compounds, suggesting their utilization under a bio-economic and environmental strategic framework.

Keywords: Antioxidant activity, grape skin, phenolic compounds, waste recovery.

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Authors: Mardiana Ahamad Zabidi, Nurain Abdul Karim, Nur Shazrinna Sazali

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Mangosteen (Garcinia mangostana) pericarp is considered as agricultural waste and not fully utilized in food products. It is widely reported that mangosteen pericarp contains high antioxidant properties. The objective of this study is to develop novel yellow alkaline noodle (YAN) substituted with different levels of mangosteen pericarp powder (MPP). YAN formulation was substituted with different levels of MPP (0%, 5%, 10% and 15%). The effect on nutritional and antioxidant properties was evaluated. Higher substitution levels of MPP resulted in significant increase (p<0.05) of ash, fibre, specific mineral elements and antioxidant properties (total phenolic, total flavonoid, anthocyanin and DPPH) than control sample.

Keywords: Yellow alkaline noodle, mangosteen pericarp powder, proximate composition, antioxidant properties.

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Authors: R. Razavizadeh, F. Adabavazeh, M. Rezaee Chermahini

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Salinity is one of the most widespread agricultural problems in arid and semi-arid areas that limits the plant growth and crop productivity. In this study, the salt stress effects on protein, reducing sugar, proline contents and antioxidant enzymes activities of Carum copticum L. under in vitro conditions were studied. Seeds of C. copticum were cultured in Murashige and Skoog (MS) medium containing 0, 25, 50, 100 and 150 mM NaCl and calli were cultured in MS medium containing 1 μM 2, 4-dichlorophenoxyacetic acid, 4 μM benzyl amino purine and different levels of NaCl (0, 25, 50, 100 and 150 mM). After NaCl treatment for 28 days, the proline and reducing sugar contents of shoots, roots and calli increased significantly in relation to the severity of the salt stress. The highest amount of proline and carbohydrate were observed at 150 and 100 mM NaCl, respectively. The reducing sugar accumulation in shoots was the highest as compared to roots, whereas, proline contents did not show any significant difference in roots and shoots under salt stress. The results showed significant reduction of protein contents in seedlings and calli. Based on these results, proteins extracted from the shoots, roots and calli of C. copticum treated with 150 mM NaCl showed the lowest contents. The positive relationships were observed between activity of antioxidant enzymes and the increase in stress levels. Catalase, ascorbate peroxidase and superoxide dismutase activity increased significantly under salt concentrations in comparison to the control. These results suggest that the accumulation of proline and sugars, and activation of antioxidant enzymes play adaptive roles in the adaptation of seedlings and callus of C. copticum to saline conditions.

Keywords: Antioxidant enzymes, Carum copticum, organic solutes, salt stress.

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Authors: Mohamed H. Khalifa, Fikry I. El-Shahawi, Nabil A. Mansour

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The cotton leafworm, Spodoptera littoralis (Boisduval) is a major insect pest of vegetables and cotton crops in Egypt, and exhibits different levels of tolerance to certain insecticides. Chlorantraniliprole has been registered recently in Egypt for control this insect. The susceptibilities of three S. littoralis populations collected from El Behaira governorate, north Egypt to chlorantraniliprole were determined by leaf-dipping technique on 4^th instar larvae. Obvious variation of toxicity was observed among the laboratory susceptible, and three field populations with LC₅₀ values ranged between 1.53 µg/ml and 6.22 µg/ml. However, all the three field populations were less susceptible to chlorantraniliprole than a laboratory susceptible population. The most tolerant populations were sampled from El Delengat (ED) Province where S. littoralis had been frequently challenged by insecticides. Certain enzyme activity assays were carried out to be correlated with the mechanism of the observed field population tolerance. All field populations showed significantly enhanced activities of detoxification enzymes compared with the susceptible strain. The regression analysis between chlorantraniliprole toxicities and enzyme activities revealed that the highest correlation is between α-esterase or β-esterase (α-β-EST) activity and collected field strains susceptibility, otherwise this correlation is not significant (P > 0.05). Synergism assays showed the ED and susceptible strains could be synergized by known detoxification inhibitors such as piperonyl butoxide (PBO), triphenyl phosphate (TPP) and diethyl-maleate (DEM) at different levels (1.01-8.76-fold and 1.09-2.94 fold, respectively), TPP showed the maximum synergism in both strains. The results show that there is a correlation between the enzyme activity and tolerance, and carboxylic-esterase (Car-EST) is likely the main detoxification mechanism responsible for tolerance of S. littoralis to chlorantraniliprole.

Keywords: Chlorantraniliprole, detoxification enzymes, Egypt, Spodoptera littoralis.

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Authors: M. Mohamadi, A. M. Maskooki., S. A. Mortazavi

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The quality and shelf life of foods of containing lipids (fats and oils) significantly reduces due to rancidity.Applications of natural antioxidants are one of the most effective manners to prevent the oxidation of oils and lipids. The antioxidant properties of juice extracted from barberry fruit (Berberris vulgaris.L) using maceration and SWE (10 bars and 120 - 180°C) methods were investigated and compared with conventional method. The amount of phenolic compound and reduction power of all samples were determined and the data were statistically analyzed using multifactor design. The results showed that the total amount of phenolic compound increased with increasing of pressure and temprature from 1861.9 to 2439.1 (mg Gallic acid /100gr Dry matter). The ability of reduction power of SWE obtained antioxidant extract compared with BHA (synthetic antioxidant) and ascorbic acid (natural antioxidant). There were significant differences among reduction power of extracts and there were remarkable difference with BHA and Ascorbic acid (P<0.01).

Keywords: Subcritical water, Antioxidant, Barberry, Phenolic compound, Reduction power

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Authors: Marcela Capcarova, Adriana Kolesarova, Marina Medvedova, Peter Petruska, Alexander V. Sirotkin

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The aim of this study was to determine the activity of superoxide dismutase (SOD), glutathione peroxidase (GPx), total antioxidant status (TAS) and accumulation of Hsp70 in porcine ovarian granulosa cells after deoxynivalenol (DON) and zearalenone (ZEA) exposure in vitro. Porcine ovarian granulosa cells were incubated with DON/ZEA administrations as follows: group A (10/10 ng/mL), group B (100/100 ng/mL), group C (1000/1000 ng/mL), and the control group without any additions for 24h. In this study mycotoxins developed stress reaction of porcine ovarian granulosa cells and increased accumulation of Hsp70 what resulted in increasing activities of SOD and GPx in groups with lower doses of mycotoxins. High dose of DON and ZEA had opposite effect on GPx activity than the lower doses. Slight increase in TAS of porcine granulosa cells was observed after mycotoxins exposure. These results contribute towards the understanding of cellular stress and its response.

Keywords: Deoxynivalenol, zearalenone, antioxidants, Hsp70, granulosa cells.

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Authors: Maryam Azizkhani, Parvin Zandi

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Application of synthetic antioxidants such as tertbutylhydroquinon (TBHQ), in spite of their efficiency, is questioned because of their possible carcinogenic effect. The purpose of this study was application of mixtures of natural antioxidants that provide the best oxidative stability for margarine. Antioxidant treatments included 10 various mixtures (F1- F10) containing 100-500ppm tocopherol mixture (Toc), 100-200ppm ascorbyl palmitate (AP), 100- 200ppm rosemary extract (Ros) and 1000ppm lecithin(Lec) along with a control or F0 (with no antioxidant) and F11 with 120ppm TBHQ. The effect of antioxidant mixtures on the stability of margarine samples during oven test (60°C), rancimat test at 110°C and storage at 4°C was evaluated. Final ranking of natural antioxidant mixtures was as follows: F2,F10>F5,F9>F8>F1,F3,F4>F6, F7. Considering the results of this research and ranking criteria, F2(200ppmAp + 200ppmRos) and F10(200ppmRos + 200ppmToc +1000ppmLec) were recommended as substitutes for TBHQ to maintain the quality and increase the shelf-life of margarine.

Keywords: Margarine, Natural antioxidant, Oxidative stability, Shelf-life.

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Authors: L. N. Corîci, A. E. Frissen, D -J. Van Zoelen, I. F. Eggen, F. Peter, C. M. Davidescu, C. G. Boeriu

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Two commercial proteases from Bacillus licheniformis (Alcalase 2.4 L FG and Alcalase 2.5 L, Type DX) were screened for the production of Z-Ala-Phe-NH2 in batch reaction. Alcalase 2.4 L FG was the most efficient enzyme for the C-terminal amidation of Z-Ala-Phe-OMe using ammonium carbamate as ammonium source. Immobilization of protease has been achieved by the sol-gel method, using dimethyldimethoxysilane (DMDMOS) and tetramethoxysilane (TMOS) as precursors (unpublished results). In batch production, about 95% of Z-Ala-Phe-NH2 was obtained at 30°C after 24 hours of incubation. Reproducibility of different batches of commercial Alcalase 2.4 L FG preparations was also investigated by evaluating the amidation activity and the entrapment yields in the case of immobilization. A packed-bed reactor (0.68 cm ID, 15.0 cm long) was operated successfully for the continuous synthesis of peptide amides. The immobilized enzyme retained the initial activity over 10 cycles of repeated use in continuous reactor at ambient temperature. At 0.75 mL/min flow rate of the substrate mixture, the total conversion of Z-Ala-Phe-OMe was achieved after 5 hours of substrate recycling. The product contained about 90% peptide amide and 10% hydrolysis byproduct.

Keywords: packed-bed reactor, peptide amide, protease, sol-gel immobilization.

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Authors: R. Charoen, S. Tipkanon, W. Savedboworn, N. Phonsatta, A. Panya

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Rice bran protein hydrolysates (RBPH) were prepared from defatted rice bran of two different Thai rice cultivars (Plai-Ngahm-Prachinburi; PNP and Khao Dok Mali 105; KDM105) using an enzymatic method. This research aimed to optimize enzyme-assisted protein extraction. In addition, the functional properties of RBPH and their stabilities to environmental stresses including pH (3 to 8), ionic strength (0 mM to 500 mM) and the thermal treatment (30 °C to 90 °C) were investigated. Results showed that enzymatic process for protein extraction of defatted rice bran was as follows: enzyme concentration 0.075 g/ 5 g of protein, extraction temperature 50 °C and extraction time 4 h. The obtained protein hydrolysate powders had a degree of hydrolysis (%) of 21.05% in PNP and 19.92% in KDM105. The solubility of protein hydrolysates at pH 4-6 was ranged from 27.28-38.57% and 27.60-43.00% in PNP and KDM105, respectively. In general, antioxidant activities indicated by total phenolic content, FRAP, ferrous ion-chelating (FIC), and 2,2’-azino-bis-3-ethylbenzthiazoline-6-sulphonic acid (ABTS) of KDM105 had higher than PNP. In terms of functional properties, the emulsifying activity index (EAI) was was 8.78 m²/g protein in KDM105, whereas PNP was 5.05 m²/g protein. The foaming capacity at 5 minutes (%) was 47.33 and 52.98 in PNP and KDM105, respectively. Glutamine, Alanine, Valine, and Leucine are the major amino acid in protein hydrolysates where the total amino acid of KDM105 gave higher than PNP. Furthermore, we investigated environmental stresses on the stability of 5% oil in water emulsion (5% oil, 10 mM citrate buffer) stabilized by RBPH (3.5%). The droplet diameter of emulsion stabilized by KDM105 was smaller (d < 250 nm) than produced by PNP. For environmental stresses, RBPH stabilized emulsions were stable at pH around 3 and 5-6, at high salt (< 400 mM, pH 7) and at temperatures range between 30-50°C.

Keywords: Functional properties, oil in water emulsion, protein hydrolysates, rice bran protein.

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Authors: Raquel P. F. Guiné, Ana R. B. Figueiredo, Paula M. R. Correia, Fernando J. Gonçalves

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The purpose of the present work was to develop an innovative food product with nutritional properties as well as appealing organoleptic qualities. The product, a jam, was prepared with the beans’ cooking water combined with fresh apple or carrot, without the addition of any conservatives. Three different jams were produced: bean and carrot, bean and apple and bean, apple and cinnamon. The developed products underwent a sensorial analysis that revealed that the bean, apple and cinnamon jam was globally better accepted. However, with this study, the consumers determined that the bean and carrot jam had the most attractive color and the bean and apple jam the better consistency. Additionally, it was possible to analyze the jams for their chemical components, namely fat, fiber, protein, sugars and antioxidant activity. The obtained results showed that the bean and carrot jam had the highest lipid content, while the bean, apple and cinnamon jam had the highest fiber content, when compared to the other two jams. Regarding the sugar content, both jams with apple revealed similar sugar values, which were higher than the sugar content of the bean and carrot jam. The antioxidant activity was on average 10 mg TE/g.

Keywords: Bean jam, chemical composition, sensorial analysis, product acceptability.

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Authors: Chlopicka Joanna, Barton Henryk, Kryczyk Jadwiga, Francik Renata

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Recent studies demonstrated that high-fat diet increases oxidative stress in plasma and in a variety of tissues. Many researchers have been looking for natural products, which can reverse the effect of high fat diet. Recently, buckwheat is becoming common ingredient in functional food because of it properties. In study on buckwheat, it is known that, this plant plays roles as anti-oxidative, anti-inflammatory and anti-hypertensive. Nevertheless still little is known about buckwheat groats. The aim of this study was to investigate the effects of addition of buckwheat groats to the fat diet (30% lard), on some antioxidant and oxidant stress parameters in plasma and selected tissues in Wistar rats. The experiment was carried out with three months old male Wistar rats ca. 250g of body weight fed for 5 weeks with either a high-fat (30% of lard) diet or control diet, with or without addition of buckwheat groats. In plasma biochemistry and the activities of the antioxidant enzymes were measured selected tissues: glutathione peroxidase (GPX), catalase (CAT) and the levels of total and reduced glutathione (GSH), free thiol groups (pSH), antioxidant potential of plasma (FRAP) and oxidant stress indices - proteins carbonyl groups (CO) and malonyldialdehyde concentration (MDA). Activity of catalase (CAT) in plasma of rats was significantly increased in buckwheat groats groups and activity of GPx3 in plasma of rats was decreased in buckwheat groups as compared to control group. The reduced glutathione (GSH) in plasma of rats was significantly increased and protein CO was significantly decreased in buckwheat groups as compared to controls. The lowered concentration of GSH was found in serum of rats fed buckwheat groats addition but it accompanied in 7-fold increase in reduced-to-oxidized glutatione ratio, significant increase in HDL and decrease in nonHDL concentration. Conclusions: Buckwheat groats indicate a beneficial effect in inhibiting protein and lipid peroxidation in rats and improved lipid profile. These results suggest that buckwheat groats exert a significant antioxidant potential and may be used as normal food constituent to ameliorate the oxidant-induced damage in organism. 

Keywords: Antioxidant, buckwheat, high-fat diet, rats.

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Authors: Fazlollah Moosavinasab, Zhila Motamedi

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An experiment was conducted to study the effects of different types of probiotic on Sucrase enzyme activity of the small intestine mucosa in male broilers. The experimental design was arranged as randomized completely blocks in 4 × 2 factorial arrangement of treatment. 180 male broilers of Ross 308 commercial hybrid were designated into 4 groups. Three replicates of 15 birds were assigned to each treatment. Control treatments (diet contained no probiotic) were fed according to the NRC as base diet and three treatment groups were fed from the same diet plus three different types of probiotics. Birds were slaughtered after 21 and 42 days and different segments of small intestine (at 1,10,30,50,70 and 90% of total length the small intestine) were taken from each replicates (N=2) Sucrase enzyme activities were measured and recorded. Obtained data were analyzed by Spss (P<0.05). In three treatment groups, probiotic had no significant effect on sucrase activity in different ages and segments of small intestine (P<0.05). These data suggested that probiotics administration had no significant effect on treatments comparing to the control group.

Keywords: Broiler, Chicks, Probiotics, Small Intestine, Sucrase

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Authors: M. D. S. Siti-Normah, S. Sabiha-Hanim, A. Noraishah

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Agricultural residue such as oil palm fronds (OPF) is cheap, widespread and available throughout the year. Hemicelluloses extracted from OPF can be hydrolyzed to their monomers and used in production of xylooligosaccharides (XOs). The objective of the present study was to optimize the enzymatic hydrolysis process of OPF hemicellulose by varying pH, temperature, enzyme and substrate concentration for production of XOs. Hemicelluloses was extracted from OPF by using 3 M potassium hydroxide (KOH) at temperature of 40°C for 4 hrs and stirred at 400 rpm. The hemicellulose was then hydrolyzed using Trichoderma longibrachiatum xylanase at different pH, temperature, enzyme and substrate concentration. XOs were characterized based on reducing sugar determination. The optimum conditions to produced XOs from OPF hemicellulose was obtained at pH 4.6, temperature of 40°C , enzyme concentration of 2 U/mL and 2% substrate concentration. The results established the suitability of oil palm fronds as raw material for production of XOs.

Keywords: Hemicellulose, oil palm fronds, Trichoderma longibrachiatum, xylooligosaccharides.

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Authors: S. Mat Radzi, N. J. Abd Rahman, H. Mohd Noor, N. Ariffin

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Ferulic acid has widespread industrial potential by virtue of its antioxidant properties. However, it is partially soluble in aqueous media, limiting their usefulness in oil-based processes in food, cosmetic, pharmaceutical, and material industry. Therefore, modification of ferulic acid should be made by producing of more lipophilic derivatives. In this study, a preliminary investigation of lipase-catalyzed trans-esterification reaction of ethyl ferulate and olive oil was investigated. The reaction was catalyzed by immobilized lipase from Candida antarctica (Novozym 435), to produce ferulate ester, a sunscreen agent. A statistical approach of Response surface methodology (RSM) was used to evaluate the interactive effects of reaction temperature (40-80°C), reaction time (4-12 hours), and amount of enzyme (0.1-0.5 g). The optimum conditions derived via RSM were reaction temperature 60°C, reaction time 2.34 hours, and amount of enzyme 0.3 g. The actual experimental yield was 59.6% ferulate ester under optimum condition, which compared well to the maximum predicted value of 58.0%.

Keywords: Ferulic acid, Enzymatic Synthesis, Esters, RSM.

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Authors: Heri Hermansyah, Annisa Kurnia, A. Vania Anisya, Adi Surjosatyo, Yopi Sunarya, Rita Arbianti, Tania Surya Utami

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Lipase is one of biocatalyst which is applied commercially for the process in industries, such as bioenergy, food, and pharmaceutical industry. Nowadays, biocatalysts are preferred in industries because they work in mild condition, high specificity, and reduce energy consumption (high pressure and temperature). But, the usage of lipase for industry scale is limited by economic reason due to the high price of lipase and difficulty of the separation system. Immobilization of lipase is one of the solutions to maintain the activity of lipase and reduce separation system in the process. Therefore, we conduct a study about lipase immobilization with the adsorption-cross linking method using glutaraldehyde because this method produces high enzyme loading and stability. Lipase is immobilized on different kind of resin with the various functional group. Highest enzyme loading (76.69%) was achieved by lipase immobilized on anion macroporous which have anion functional group (OH^‑). However, highest activity (24,69 U/g support) through olive oil emulsion method was achieved by lipase immobilized on anion macroporous-chitosan which have amino (NH₂) and anion (OH^-) functional group. In addition, it also success to produce biodiesel until reach yield 50,6% through interesterification reaction and after 4 cycles stable 63.9% relative with initial yield. While for Aspergillus, niger lipase immobilized on anion macroporous-kitosan have unit activity 22,84 U/g resin and yield biodiesel higher than commercial lipase (69,1%) and after 4 cycles stable reach 70.6% relative from initial yield. This shows that optimum functional group on support for immobilization with adsorption-cross linking is the support that contains amino (NH₂) and anion (OH^-) functional group because they can react with glutaraldehyde and binding with enzyme prevent desorption of lipase from support through binding lipase with a functional group on support.

Keywords: Adsorption-Cross linking, lipase, resin, immobilization.

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Authors: I. Savitskaya, A. Kistaubayeva, A. Zhubanova, I. Blavachinskaiya, D. Ibrayeva, M. Abdulzhanova, A. Otarbay, A.Isabekova

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This study was conducted for the investigation of number of cellulolytic bacteria and their ability in decomposition. Seven samples surface soil were collected on cellulose Zailiskii Alatau slopes. Cellulolitic activity of new strains of Bacillus, isolated from soil is determined. Isolated cellulose degrading bacteria were screened for determination of the highest cellulose activity by quantitative assay using Congo red, gravimetric assay and colorimetric DNS method trough of the determination of the parameters of sugar reduction. Strains are assigned to: B.subtilis, B.licheniformis, B. cereus and, В. megaterium. Bacillus strains consisting of several different types of cellulases have broad substrate specificity of cellulase complexes formed by them. Cellulolitic bacteria were recorded to have highest cellulase activity and selected for optimization of cellulase enzyme production.

Keywords: Cellulose-degrading bacteria, cellulase complex, foothills soil, screening.

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Authors: Ismail I. Abo Ghanema, Kadry M. Sadek

Abstract:

Oxidative stress and overwhelming free radicals associated with diabetes mellitus are likely to be linked with development of certain complication such as retinopathy, nephropathy and neuropathy. Treatment of diabetic subjects with antioxidant may be of advantage in attenuating these complications. Olive leaf (Oleaeuropaea), has been endowed with many beneficial and health promoting properties mostly linked to its antioxidant activity. This study aimed to evaluate the significance of supplementation of Olive leaves extract (OLE) in reducing oxidative stress, hyperglycemia and hyperlipidemia in Sterptozotocin (STZ)- induced diabetic rats. After induction of diabetes, a significant rise in plasma glucose, lipid profiles except High density lipoproteincholestrol (HDLc), malondialdehyde (MDA) and significant decrease of plasma insulin, HDLc and Plasma reduced glutathione GSH as well as alteration in enzymatic antioxidants was observed in all diabetic animals. During treatment of diabetic rats with 0.5g/kg body weight of Olive leaves extract (OLE) the levels of plasma (MDA) ,(GSH), insulin, lipid profiles along with blood glucose and erythrocyte enzymatic antioxidant enzymes were significantly restored to establish values that were not different from normal control rats. Untreated diabetic rats on the other hand demonstrated persistent alterations in the oxidative stress marker (MDA), blood glucose, insulin, lipid profiles and the antioxidant parameters. These results demonstrate that OLE may be of advantage in inhibiting hyperglycemia, hyperlipidemia and oxidative stress induced by diabetes and suggest that administration of OLE may be helpful in the prevention or at least reduced of diabetic complications associated with oxidative stress.

Keywords: Diabetes mellitus, olive leaves, oxidative stress, red blood cells

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Authors: A. M. Tarola, R. Preti, A. M. Girelli, P. Campana

Abstract:

Safeguarding, studying and enhancing biodiversity play an important and indispensable role in re-launching agriculture. The ancient local varieties are therefore a precious resource for genetic and health improvement. In order to protect biodiversity through the recovery and valorization of autochthonous varieties, in this study we analyzed 12 samples of four ancient apple cultivars representative of Friuli Venezia Giulia, selected by local farmers who work on a project for the recovery of ancient apple cultivars. The aim of this study is to evaluate the polyphenolic profile and the antioxidant capacity that characterize the organoleptic and functional qualities of this fruit species, besides having beneficial properties for health. In particular, for each variety, the following compounds were analyzed, both in the skins and in the pulp: gallic acid, catechin, chlorogenic acid, epicatechin, caffeic acid, coumaric acid, ferulic acid, rutin, phlorizin, phloretin and quercetin to highlight any differences in the edible parts of the apple. The analysis of individual phenolic compounds was performed by High Performance Liquid Chromatography (HPLC) coupled with a diode array UV detector (DAD), the antioxidant capacity was estimated using an in vitro essay based on a Free Radical Scavenging Method and the total phenolic compounds was determined using the Folin-Ciocalteau method. From the results, it is evident that the catechins are the most present polyphenols, reaching a value of 140-200 μg/g in the pulp and of 400-500 μg/g in the skin, with the prevalence of epicatechin. Catechins and phlorizin, a dihydrohalcone typical of apples, are always contained in larger quantities in the peel. Total phenolic compounds content was positively correlated with antioxidant activity in apple pulp (r² = 0,850) and peel (r² = 0,820). Comparing the results, differences between the varieties analyzed and between the edible parts (pulp and peel) of the apple were highlighted. In particular, apple peel is richer in polyphenolic compounds than pulp and flavonols are exclusively present in the peel. In conclusion, polyphenols, being antioxidant substances, have confirmed the benefits of fruit in the diet, especially as a prevention and treatment for degenerative diseases. They demonstrated to be also a good marker for the characterization of different apple cultivars. The importance of protecting biodiversity in agriculture was also highlighted through the exploitation of native products and ancient varieties of apples now forgotten.

Keywords: Apple, biodiversity, polyphenols, antioxidant activity, HPLC-DAD, characterization.

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Authors: Hazim Saadoon Aljewari, Mohammed Ibraheem Nader, Abdul Hussain M. Alfaisal, NatthidaWeerapreeyakul, Sahapat

Abstract:

L-asparaginase was extracted from pathogenic Escherichia coli which was isolated from urinary tract infection patients. L-asparaginase was purified 96-fold by ultrafiltration, ion exchange and gel filtration giving 39.19% yield with final specific activity of 178.57 IU/mg. L-asparaginase showed 138,356±1,000 Dalton molecular weight with 31024±100 Dalton molecular mass. Kinetic properties of enzyme resulting 1.25×10-5 mM Km and 2.5×10-3 M/min Vmax. L-asparaginase showed a maximum activity at pH 7.5 when incubated at 37 ºC for 30 min and illustrated its full activity (100%) after 15 min incubation at 20-37 ºC, while 70% of its activity was lost when incubated at 60 ºC. L-asparaginase showed cytotoxicity to U937 cell line with IC50 0.5±0.19 IU/ml, and selectivity index (SI=7.6) about 8 time higher selectivity over the lymphocyte cells. Therefore, the local pathogenic E. coli strains may be used as a source of high yield of L-asparaginase to produce anti cancer agent with high selectivity.

Keywords: L-asparaginase, Purification, Cytotoxicity, selectivity index

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Authors: Supriyono, Sumardiyono, Rendy J. Pramono

Abstract:

Biodiesel is one of the alternative fuels promising for substituting petrodiesel as energy source which has an advantage as it is sustainable and eco-friendly. Due to the raw material that tends to decompose during storage, biodiesel also has the same characteristic that tends to decompose during storage. Biodiesel decomposition will form higher acid value as the result of oxidation to double bond on a fatty acid compound on biodiesel. Thus, free fatty acid value could be used to evaluate degradation of biodiesel due to the oxidation process. High free fatty acid on biodiesel could impact on the engine performance. Decomposition of biodiesel due to oxidation reaction could prevent by introducing a small amount of antioxidant. The origin of raw materials and the process for producing biodiesel will determine the effectiveness of antioxidant. Biodiesel made from high free fatty acid (FFA) crude palm oil (CPO) by using two steps esterification is vulnerable to oxidation process which is resulted in increasing on the FFA value. Tocopherol also known as vitamin E is one of the antioxidant that could improve the stability of biodiesel due to decomposition by the oxidation process. Tocopherol 0.5% concentration on palm oil biodiesel could reduce 13% of increasing FFA under temperature 80 °C and exposing time 180 minute.

Keywords: Antioxidant, biodiesel, decomposition, oxidation, tocopherol.

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Authors: Supriyono, Sumardiyono, Rendy J. Pramono

Abstract:

Biodiesel is one of the alternative fuels promising for substituting petrodiesel as energy source which has an advantage as it is sustainable and eco-friendly. Due to the raw material that tends to decompose during storage, biodiesel also has the same characteristic that tends to decompose during storage. Biodiesel decomposition will form higher acid value as the result of oxidation to double bond on a fatty acid compound on biodiesel. Thus, free fatty acid value could be used to evaluate degradation of biodiesel due to the oxidation process. High free fatty acid on biodiesel could impact on the engine performance. Decomposition of biodiesel due to oxidation reaction could prevent by introducing a small amount of antioxidant. The origin of raw materials and the process for producing biodiesel will determine the effectiveness of antioxidant. Biodiesel made from high free fatty acid (FFA) crude palm oil (CPO) by using two steps esterification is vulnerable to oxidation process which is resulted in increasing on the FFA value. Tocopherol also known as vitamin E is one of the antioxidant that could improve the stability of biodiesel due to decomposition by the oxidation process. Tocopherol 0.5% concentration on palm oil biodiesel could reduce 13% of increasing FFA under temperature 80 °C and exposing time 180 minute.

Keywords: Antioxidant, biodiesel, decomposition, oxidation, tocopherol.

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Authors: Simon Brown, Noorzaid Muhamad, David C Simcock

Abstract:

The kinetic properties of enzymes are often reported using the apparent KM and Vmax appropriate to the standard Michaelis-Menten enzyme. However, this model is inappropriate to enzymes that have more than one substrate or where the rate expression does not apply for other reasons. Consequently, it is desirable to have a means of estimating the appropriate kinetic parameters from the apparent values of KM and Vmax reported for each substrate. We provide a means of estimating the range within which the parameters should lie and apply the method to data for glutamate dehydrogenase from the nematode parasite of sheep Teladorsagia circumcincta.

Keywords: enzyme kinetics, glutamate dehydrogenase, intervalanalysis, parameter estimation.

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Authors: Amir Amiri, Naghmeh Morakabati

Abstract:

During the recent years the six-fold growth of cancer in Iran has led the production of healthy products to become a challenge in the food industry. Due to the young population in the country, the consumption of fast foods is growing. The chemical cancer-causing preservatives are used to produce these products more than the standard; so using an appropriate alternative seems to be important. On the one hand, the plant essential oils show the high antimicrobial potential against pathogenic and spoilage microorganisms and on the other hand they are highly volatile and decomposed under the processing conditions. The study aims to produce the loaded chitosan nanoparticles with different concentrations of savory essential oil to improve the anti-microbial property and increase the resistance of essential oil to oxygen and heat. The encapsulation efficiency was obtained in the range of 32.07% to 39.93% and the particle size distribution of the samples was observed in the range of 159 to 210 nm. The range of Zeta potential was obtained between -11.9 to -23.1 mV. The essential oil loaded in chitosan showed stronger antifungal activity against Rhizopus stolonifer. The results showed that the antioxidant property is directly related to the concentration of loaded essential oil so that the antioxidant property increases by increasing the concentration of essential oil. In general, it seems that the savory essential oil loaded in chitosan particles can be used as a food processor.

Keywords: Chitosan, encapsulation, essential oil, nanogel.

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Authors: Malinee Pongsavee

Abstract:

Sodium formate is the chemical substance used for food additive. Catalase is the important antioxidative enzyme in protecting the cell from oxidative damage by reactive oxygen species (ROS). The resultant level of oxidative stress in sodium formatetreated lymphocytes was investigated. The sodium formate concentrations of 0.05, 0.1, 0.2, 0.4 and 0.6 mg/mL were treated in human lymphocytes for 12 hours. After 12 treated hours, catalase activity change was measured in sodium formate-treated lymphocytes. The results showed that the sodium formate concentrations of 0.4 and 0.6 mg/mL significantly decreased catalase activities in lymphocytes (P < 0.05). The change of catalase activity in sodium formate-treated lymphocytes may be the oxidative damage marker for detect sodium formate exposure in human.

Keywords: Sodium formate, catalase activity, oxidative damage marker, toxicity.

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