Search results for: glutathione conjugating enzymes

Authors: Seyed Hassan Moosa-kazemi, Hassan Vatandoost

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Malaria, Dirofilaria immitis (dog heart worm), and D. repens (dirofilariasis), which are transmitted by mosquitoes, have been reported in Iran. The Iranian mosquito fauna includes seven genera, 65 species, and three subspecies. Aedes albopictus has been reported since. West Nile, Sindbis, Dengue, Japanese encephalitis viruses, and the nematode Setaria (setariasis) has been reported in the country but there are no information about their vectors in Iran. Iran is malaria elimination phase. Insecticides residual spraying (IRS), distributed of insecticides long lasting treated nets (ITNs), fogging, release of larvivours fishes and Bacillus thuringiensis, chemical larviciding, as well as case finding and manipulation and modification of breeding places carried out thought the IVM program in the country. Prolonged exposure to insecticides over several generations of the vectors, develop resistance, a capacity to survive contact with insecticides. However, use of insecticides in agriculture has often been implicated as contributing to resistance in mosquito’s vectors. Resistance of mosquitoes to some insecticides has been documented just within a few years after the insecticides were introduced. Some enzymes such as monooxygenases, esterases and glutathione S-transferases have been considered as a reason for resistance to pyrethroid insecticides. In conclusion, regarding to documented resistance and tolerance of mosquitoes vectors to some insecticides, resistance management is suggested by using new insecticide with novel mode of action.

Keywords: control, Iran, resistance, vector

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Authors: Nivedita Sharma

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The global demand for fossil fuels is very high, but their use is not sustainable since its reserves are declining. Additionally, fossil fuels are responsible for the accumulation of greenhouse gases. The emission of greenhouse gases from the transport sector can be reduced by substituting fossil fuels by biofuels. Thus, renewable fuels capable of sequestering carbon dioxide are in high demand. Second‐generation biofuels, which require lignocellulosic biomass as a substrate and ultimately producing ethanol, fall largely in this category. Bioethanol is a favorable and near carbon-neutral renewable biofuel leading to reduction in tailpipe pollutant emission and improving the ambient air quality. Lignocellulose consists of three main components: cellulose, hemicellulose and lignin which can be converted to ethanol with the help of microbial enzymes. Enzymatic hydrolysis of lignocellulosic biomass in 1st step is considered as the most efficient and least polluting methods for generating fermentable hexose and pentose sugars which subsequently are fermented to power alcohol by yeasts in 2nd step of the process. In the present technology, a complete bioconversion process i.e. potential hydrolytic enzymes i.e. cellulase and xylanase producing microorganisms have been isolated from different niches, screened for enzyme production, identified using phenotyping and genotyping, enzyme production, purification and application of enzymes for saccharification of different lignocellulosic biomass followed by fermentation of hydrolysate to ethanol with high yield is to be presented in detail.

Keywords: cellulase, xylanase, lignocellulose, bioethanol, microbial enzymes

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Authors: Rajnish Gupta, R. S. Gupta

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Present research was aimed to investigate antidiabetic and antioxidant status of Lupeol in streptozotocin induced diabetes. Rats were divided into following groups mainly: control, diabetic, normal group as well as diabetic treated with Lupeol at 25 and 35 mg/kg b.wt./day for 21 days, diabetic group treated with glibenclamide. Tissue (pancreas, kidney and liver) as well as serum biochemical parameters were analysed for any abnormal behavior. Lupeol administration reduced diabetes onset with significant improvement in serum insulin level also strengthened by increase in β-Cell counts. A significant decrease was observed in serum glucose level. Furthermore, Lupeol treatment increased the antioxidant enzymes, glycolytic enzymes and also protein levels with a decrease in the level of thiobarbituric acid-reactive oxygen species and gluconeogenic enzymes. Present study proves that Lupeol administration significantly reinstated serum and tissue biochemical parameters and thus strengthening its antidiabetic potential.

Keywords: oxidative stress, pterostilbene, thiobarbituric acid, reactive oxygen species

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Authors: Manal E. A Elhalwagy, Nadia Amin Abdulmajeed, Hanan S. Alnahdi, Enas N. Danial

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Baron is herbicide includes (48% glyphosate) widely used in Egypt. The present study assesses the cytotoxic and genotoxic effect of baron on rats liver. Two groups of rats were treated orally with 1/10 LD 50, (275.49 mg kg -1) and 1/40 LD 50, (68.86 mg kg-1) glyphosate for 28 days compared with control group. Serum and liver tissues were taken at 14 and 28 days of treatment. An inhibition in Alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities were recorded at both treatment periods and reduction in total serum protein (TP) and albumin (ALB). However, non-significant changes in serum acetylcholinesterase (AChE). Elevation in oxidative stress biomarker malondyaldehyde (MDA) and the decline in detoxification biomarker total reduced glutathione (GSH), Glutathione S-transferase (GST) and superoxide dismutase (SOD) in liver tissues led to increase in percentage of DNA damage. Destruction in liver tissue architecture was observed . Although, Baron was classified in the safe category pesticides repeated exposure to small doses has great danger effect.

Keywords: glyphosate, liver toxicity, oxidative stress, DNA damage, commet assay

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Authors: R. Pravin Kumar, L. Roopa

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Enzymes are molecular machines used in various industries such as pharmaceuticals, cosmetics, food and animal feed, paper and leather processing, biofuel, and etc. Nevertheless, this has been possible only by the breath-taking efforts of the chemists and biologists to evolve/engineer these mysterious biomolecules to work the needful. Main agenda of this enzyme engineering project is to derive screening and selection tools to obtain focused libraries of enzyme variants with desired qualities. The methodologies for this research include the well-established directed evolution, rational redesign and relatively less established yet much faster and accurate insilico methods. This concept was initiated as a Receptor Rependent-4Dimensional Quantitative Structure Activity Relationship (RD-4D-QSAR) to predict kinetic properties of enzymes and extended here to study transaminase by a 7D QSAR approach. Induced-fit scenarios were explored using Quantum Mechanics/Molecular Mechanics (QM/MM) simulations which were then placed in a grid that stores interactions energies derived from QM parameters (QMgrid). In this study, the mutated enzymes were immersed completely inside the QMgrid and this was combined with solvation models to predict descriptors. After statistical screening of descriptors, QSAR models showed > 90% specificity and > 85% sensitivity towards the experimental activity. Mapping descriptors on the enzyme structure revealed hotspots important to enhance the enantioselectivity of the enzyme.

Keywords: QMgrid, QM/MM simulations, RD-4D-QSAR, transaminase

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Authors: Muralisankar Thirunavukkarasu, Saravana Bhavan Periyakali, Santhanam Perumal

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The present study was performed to assess the effects of dietary copper (Cu) on growth, biochemical constituents, digestive enzyme activities, enzymatic antioxidant and metabolic enzymes of the freshwater prawn, Macrobrachium rosenbergii post larvae (PL). The Cu was supplemented at 0, 10, 20, 40, 60 and 80 mg kg-1 with the basal diets. Cu supplemented diets were fed to M. rosenbergii PL for a period of 90 days. At the end of the feeding experiment, 40 mg kg-1 Cu supplemented feeds fed PL showed significant (P < 0.05) improvement in survival, growth, digestive enzyme activities and concentrations of biochemical constituents. However, PL fed with 60 to 80 mg Cu kg-1 showed negative performance. Activities of enzymatic antioxidants, metabolic enzymes and lipid peroxidation in the muscle and hepatopancreas showed insignificant alterations (P > 0.05) up to 40 mg kg-1 Cu supplemented feeds fed PL. Whereas, 60 and 80 mg of Cu kg-1 supplemented feeds fed PL showed significant alterations on these antioxidants and metabolic enzymes levels. It indicates that beyond 40 mg Cu kg-1 diets were produced some toxic to M. rosenbergii PL. Therefore, the present study suggests that 40 mg Cu kg-1 can be supplemented in the diets of M. rosenbergii PL for regulating better survival and growth.

Keywords: antioxidants, biochemical constituents, copper, growth, Macrobrachium rosenbergii

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Authors: Jessica Pinheiro Silva, Brenda Rabello De Camargo, Daniel Gusmao De Morais, Eliane Ferreira Noronha

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The utilization of lignocellulosic biomass as source of polysaccharides for industrial applications requires an arsenal of enzymes with different mode of action able to hydrolyze its complex and recalcitrant structure. Clostridium thermocellum is gram-positive, thermophilic bacterium producing lignocellulosic hydrolyzing enzymes in the form of multi-enzyme complex, termed celulossomes. This complex has several hydrolytic enzymes attached to a large and enzymically inactive protein known as Cellulosome-integrating protein (CipA), which serves as a scaffolding protein for the complex produced. This attachment occurs through specific interactions between cohesin modules of CipA and dockerin modules in enzymes. The present work aims to construct celulosomes in vitro with the structural protein CipA, a xylanase called Xyn10D and a cellulose called CelJ from C.thermocellum. A mini-scafoldin was constructed from modules derived from CipA containing two cohesion modules. This was cloned and expressed in Escherichia coli. The other two genes were cloned under the control of the alcohol oxidase 1 promoter (AOX1) in the vector pPIC9 and integrated into the genome of the methylotrophic yeast Pichia pastoris GS115. Purification of each protein is being carried out. Further studies regarding enzymatic activity of the cellulosome is going to be evaluated. The cellulosome built in vitro and composed of mini-CipA, CelJ and Xyn10D, can be very interesting for application in industrial processes involving the degradation of plant biomass.

Keywords: cellulosome, CipA, Clostridium thermocellum, cohesin, dockerin, yeast

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Authors: Imran Muhammad

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The main cause of infectious disease in the modern world is Mycobacterium Tuberculosis (MT). To combat tuberculosis, new and efficient drugs are an urgent need in the modern world. Schif bases are potent for their biological pharmacophore activity. Thus we selected different Vanillin-based Schiff bases for their binding activity against target enzymes of Mycobacterium tuberculosis that is (DprE1 (decaprenyl phosphoryl-β-D-ribose 2′-epimerase), and DNA gyrase subunit-A), using molecular docking. We evaluate the inhibition potential, interaction, and binding mode of these compounds with the target enzymes.

Keywords: schiff bases, tuberculosis, DNA gyrase, DprE1, docking

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Authors: Eric Beyegue, Boris Azantza, Judith Laure Ngondi, Julius E. Oben

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Background: It is clearly that phytochemical constituents of plants in relation exhibit free radical scavenging, antioxidant and glycosylation properties. This study investigated the in vitro antioxidant and free radical scavenging, inhibited activities against α-amylase and invertase enzymes of stem bark extracts C. edulis (Olacaceae). Methods: Four extracts (hexane, dichloromethane, ethanol and aqueous) from the barks of C. edulis were used in this study. Colorimetric in vitro methods were using for evaluate free radical scavenging activity DPPH, ABTS, NO, OH, antioxidant capacity, glycosylation activity, inhibition of α-amylase and invertase activities, phenolic, flavonoid and alkaloid contents. Results: C. edulis extracts (CEE) had a higher scavenging potential on the 2, 2-diphenyl-1-picrylhydrazyl (DPPH), hydroxyl (OH), nitrite oxide (NO), 2, 2-azinobis (3-ethylbenzthiazoline)-6-sulfonic acid (ABTS) radicals and glucose scavenging with the IC50 varied between 41.95 and 36694.43 µg/ml depending on the solvent of extraction. The ethanol extract of C. edulis stem bark (CE EtOH) showed the highest polyphenolic (289.10 + 30.32), flavonoid (1.12 + 0.09) and alkaloids (18.47 + 0.16) content. All the tested extracts demonstrated a relative high inhibition potential against α-amylase and invertase digestive enzymes activities. Conclusion: This study suggests that CEE exhibited higher antioxidant potential and significant inhibition potential against digestive enzymes.

Keywords: Coula edulis, antioxidant, scavenging activity, amylase, invertase

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Authors: Famuwagun A. A., Abiona O. O., Gbadamosi S.O., Adeboye O. A., Adebooye O. C.

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The need to provide more information on the perceived bioactive status of the peel of plantain fruit informed the design of this research. Matured Plantain fruits were harvested, and fruits were allowed to ripen spontaneously. Samples of plantain fruit were taken every fortnight, and the peels were removed. The peels were dried using two different drying techniques (Oven drying and sun drying) and milled into powdery forms. Other samples were picked and processed in a similar manner on the first, third, seventh and tenth day until the peels of the fruits were fully ripped, resulting in eight different samples. The anti-oxidative properties of the samples using different assays (DPPH, FRAP, MCA, HRSA, SRSA, ABTS, ORAC), inhibitory activities against enzymes related to diabetes (alpha-amylase and glucosidase) and inhibition against angiotensin-converting enzymes (ACE) were evaluated. The result showed that peels of plantain fruits on the 7th day of ripening and sundried exhibited greater inhibitions against free radicals, which enhanced its antioxidant activities, resulting in greater inhibitions against alpha-amylase and alpha-glucosidase enzymes. Also, oven oven-dried sample of the peel of plantain fruit on the 7th day of ripening had greater phenolic contents than the other samples, which also resulted in higher inhibition against angiotensin converting enzymes when compared with other samples. The results showed that even though the unripe peel of plantain fruit is assumed to contain excellent bioactive activities, consumption of the peel should be allowed to ripen for seven days after maturity and harvesting so as to derive maximum benefit from the peel.

Keywords: functional ingredient, diabetics, hypertension, functional foods

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Authors: Patsarawadee Paojinda, Waranya Imprasittichai, Sudaratana R. Krungkrai, Nirianne Marie Q. Palacpac, Toshihiro Horii, Jerapan Krungkrai

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Fusion of the last two enzymes in the pyrimidine biosynthetic pathway in the inversed order by having COOH-terminal orotate phosphoribosyltransferase (OPRT) and NH2-terminal orotidine 5'-monophosphate decarboxylase (OMPDC), as OMPDC-OPRT, are described in many organisms. Here, we produced gene fusions of Plasmodium falciparum OMPDC-OPRT and expressed the bifunctional protein in Escherichia coli. The enzyme was purified to homogeneity using affinity and anion-exchange chromatography, exhibited enzymatic activities and functioned as a dimer. The activities, although unstable, can be stabilized by its substrate and product during purification and long-term storage. Furthermore, the enzyme expressed a perfect catalytic efficiency (kcat/Km). The kcat was selectively enhanced up to 3 orders of magnitude, while the Km was not much affected and remained at low µM levels when compared to the monofunctional enzymes. The fusion of the two enzymes, creating a “super-enzyme” with perfect catalytic power and more flexibility, reflects cryptic relationship of enzymatic reactivaties and metabolic functions on molecular evolution.

Keywords: bifunctional enzyme, orotate phosphoribosyltransferase, orotidine 5'-monophosphate decarboxylase, plasmodium falciparum

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Authors: A. V. Shrirao, N. I. Kochar, A. V. Chandewar

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The flower extract of Butea monosperma herb has been used traditionally in India for medicinal purposes. The plant has been reported to treat hyperglycemia and associated hyperlipidemia. Hyperlipidemia and oxidative stress are known to accelerate coronary artery disease and progression of atherosclerotic lesions. The present work was undertaken to investigate the possible antihyperlipidemic and antioxidative effect of Butea monosperma flowers on hyperlipidemic rats. Hyperlipidemia was induced in rats by a single intraperitonial (i.p.) injection of Triton WR 1339 (400 mg/kg) and it showed sustained elevated levels of serum cholesterol and triglyceride. Ethanolic extract of Butea monosperma flowers (Et-BM) (250 and 500 mg/kg/day) was administered to normal and hyperlipidemic rats for 14 days. Serum and liver tissue were analyzed at three different time intervals for lipid profile and antioxidants enzymes and the activity were compared to the cholesterol-lowering drug, Atorvastatin (10 mg/kg). Parameters were altered during hyperlipidemia and reverted back to near normal values after Et-BM treatment or standard drug Atorvastatin. Lipid peroxidation decreased whereas the activities of superoxide dismutase, glutathione peroxidase and catalase increased in Et-BM treated rats. Pronounced changes were observed at 500 mg/kg of Et-BM for 2 weeks and it was comparable to the standard drug Atorvastatin. The current study provides strong evidence that Et-BM has a remarkable beneficial effect in treating hyperlipidemia and ROS without any side effects at the dosage and duration studied.

Keywords: antioxidant, butea monopserma, hyperlipidemia, triton WR 1339

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Authors: Abdur Rahman, Saima, T. N. Pasha, Muhammad Younus, Yassar Abbas, Shahid Jaleel

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Non-starch polysaccharides (NSPs) are not completely digested by broiler endogenous enzymes and consequently the soluble NSPs in feed results in high digesta viscosity and poor retention of nutrients. Supplementation of NSPs digesting enzymes may release the nutrients from feed and reduce the anti-nutritional effects of NSP’s. The present study was conducted to determine the effects of NSPs digesting enzymes (Zympex) in broiler chicks. A total of 120 day old broiler chicks (Hubbard) were categorized into 3 treatments and each treatment was having four replicates with 10 birds in each. Dietary treatments comprised of Basal diet (2740 KCal/Kg) as control-1 (T1), low energy diet (2630 KCal/kg) control-2 (T2) and low energy diet with 0.5 gm/Kg enzyme as T3. Multi-enzymes supplementation showed significant (P < 0.05) positive effect on weight gain (last three weeks), feed intake (last two weeks), FCR (1st, 2nd, 4th and 5th) and nutrient retention in T3 when compared with control-2. Weight gain was lower (P < 0.05) in low caloric feed group C when compared with control-1 in all weeks except last week (P > 0.05), feed consumption was significantly lower (P < 0.05) in 5th week and results showed significantly poor FCR (P < 0.05) in 2nd, 3rd and 4th week but non-significant effect in 1st and 5th week when compared with control-1 group, which revealed the positive effect of enzyme supplementation in low energy diet. These results revealed that enzyme supplementation releases more energy from low energy diets and results in equal performance to normal diet.

Keywords: body weight, FCR, feed intake, enzyme, non-starch polysaccharides

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Authors: Aman Upaganlawar, Chandrasekhar Upasani

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The present study was designed to screen the neuroprotective and antioxidant activity of corosolic acid in painful diabetic neuropathy (DN). Diabetes was induced in rats by single dose of STZ (60mg/kg, i.p). Diabetic rats were tested every week for the development of pain, at 5th week rats showed sensation of pain. At 6th week the rats developed significant neuropathic pain. They were divided into different groups and treated with Corosolic acid (2 and 4 mg/kg, p.o) for further two weeks. Pain was assessed in the diabetic rats by mechano-tactil allodynia, mechanical hyperalgesia and cold allodynia. At the end of treatment period rats were scarified and biochemical changes such as plasma glucose level, endogenous antioxidants (Lipid peroxidation, reduced glutathione, superoxide dismutase and catalase) in sciatic nerve were evaluated. Further Na+/K+ ATPase and nitric oxide content was also evaluated. Treatment with corosolic acid for two weeks restored the altered body weight and elevated blood sugar level. Further corosolic acid showed dose dependent reduction in pain in neuropathic animals. The level of endogenous antioxidants enzymes, Na+/K+ ATPase and nitric oxide were significantly prevented. In conclusion, the result of the present study suggests the antidiabetic, antioxidant and neuroprotectieve property of corosolic acid in diabetic rats with neuropathic pain.

Keywords: neuropathic pain, diabetes, corosolic acid, antioxidant

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Authors: Astorga-Trejo Rebeca, Fonseca-Peralta Héctor Manuel, Beltrán-Arredondo Laura Ivonne, Castro-Martínez Claudia

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The use of biomass as feedstock for the production of fuels and other chemicals of interest is an ever-growing accepted option in the way to the development of biorefinery complexes; in the Mexican state of Sinaloa, two million tons of residues from corn crops are produced every year, most of which can be converted to bioethanol and other products through biotechnological conversion using yeast and other microorganisms. Therefore, the objective of this work was to take advantage of corn stover and evaluate its potential as a substrate for the production of second-generation bioethanol (2G), enzymes, and xylitol. To produce bioethanol 2G, an acid-alkaline pretreatment was carried out prior to saccharification and fermentation. The microorganisms used for the production of enzymes, as well as for the production of xylitol, were isolated and characterized in our workgroup. Statistical analysis was performed using Design Expert version 11.0. The results showed that it is possible to obtain 2G bioethanol employing corn stover as a carbon source and Saccharomyces cerevisiae ItVer01 and Candida intermedia CBE002 with yields of 0.42 g and 0.31 g, respectively. It was also shown that C. intermedia has the ability to produce xylitol with a good yield (0.46 g/g). On the other hand, qualitative and quantitative studies showed that the native strains of Fusarium equiseti (0.4 IU/mL - xylanase), Bacillus velezensis (1.2 IU/mL – xylanase and 0.4 UI/mL - amylase) and Penicillium funiculosum (1.5 IU / mL - cellulases) have the capacity to produce xylanases, amylases or cellulases using corn stover as raw material. This study allowed us to demonstrate that it is possible to use corn stover as a carbon source, a low-cost raw material with high availability in our country, to obtain bioproducts of industrial interest, using processes that are more environmentally friendly and sustainable. It is necessary to continue the optimization of each bioprocess.

Keywords: biomass, corn stover, biorefinery, bioethanol 2G, enzymes, xylitol

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Authors: Madalin Enache, Simona Neagu, Roxana Cojoc, Ioana Gomoiu, Delia Ionela Dobre, Ancuta Roxana Trifoi

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Various types of halophilic and halotolerant microorganisms able to be cultivated in laboratory on culture media with a wide range of sodium chloride content are isolated from several salted environments. The extracellular enzymes of these microorganisms showed the enzymatic activity in these spectrums of salinity thus being attractive for several biotechnological processes developed at high ionic strength. In present work, a number of amylase, protease, esterase, lipase, cellulase, pectinase, xilanases and innulinase were identified for more than 50th bacterial strains isolated from water samples and sapropelic mud from four saline and hypersaline lakes located in Romanian plain. On the other hand, the cellulase and pectinase activity were also detected in some halotolerant microorganisms isolated from secondary agricultural flow of grapes processing. The preliminary data revealed that from totally tested strains seven harbor proteases activity, eight amylase activity, four for esterase and another four for lipase, three for pectinase and for one strain were identified either cellulase or pectinase activity. There were no identified enzymes able to hydrolase innulin added to culture media. Several strains isolated from sapropelic mud showed multiple extracellular enzymatic activities, namely three strains harbor three activities and another seven harbor two activities. The data revealed that amylase and protease activities were frequently detected if compare with other tested enzymes. In the case of pectinase were investigated, their ability to be used for increasing resveratrol recovery from material resulted after grapes processing. In this way, the resulted material from grapes processing was treated with microbial supernatant for several times (two, four and 24 hours) and the content of resveratrol was detected by High Performance Liquid Chromatography method (HPLC). The preliminary data revealed some positive results of this treatment.

Keywords: halophilic microorganisms, enzymes, pectinase, salinity

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Authors: Neeraj Neeraj, Soumen Basu, Banibrata Maity

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Glutathione (GSH) is one of the most important biomolecules having small molecular weight, which helps in various cellular functions like regulation of gene, xenobiotic metabolism, preservation of intracellular redox activities, signal transduction, etc. Therefore, the detection of GSH requires huge attention by using extremely selective and sensitive techniques. Herein, a rapid fluorometric nanosensor is designed by combining carbon dots (Cdots) and MnO₂ nanoparticles of different sizes for the detection of GSH. The bottom-up approach, i.e., microwave method, was used for the preparation of the water soluble and greatly fluorescent Cdots by using ascorbic acid as a precursor. MnO₂ nanospheres of different sizes (large, medium, and small) were prepared by varying the ratio of concentration of methionine and KMnO₄ at room temperature, which was confirmed by HRTEM analysis. The successive addition of MnO₂ nanospheres in Cdots results fluorescence quenching. From the fluorescence intensity data, Stern-Volmer quenching constant values (KS-V) were evaluated. From the fluorescence intensity and lifetime analysis, it was found that the degree of fluorescence quenching of Cdots followed the order: large > medium > small. Moreover, fluorescence recovery studies were also performed in the presence of GSH. Fluorescence restoration studies also show the order of turn on follows the same order, i.e., large > medium > small, which was also confirmed by quantum yield and lifetime studies. The limits of detection (LOD) of GSH in presence of Cdots@different sized MnO₂ nanospheres were also evaluated. It was observed thatLOD values were in μM region and lowest in case of large MnO₂ nanospheres. The separation distance (d) between Cdots and the surface of different MnO₂ nanospheres was determined. The d values increase with increase in the size of the MnO₂ nanospheres. In summary, the synthesized Cdots@MnO₂ nanocomposites acted as a rapid, simple, economical as well as environmental-friendly nanosensor for the detection of GSH.

Keywords: carbon dots, fluorescence, glutathione, MnO₂ nanospheres, turn off-on

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Authors: Irina E. Sukovataya, Oleg S. Sutormin, Valentina A. Kratasyuk

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Effect of viscosity of media on kinetic parameters of the coupled enzyme system NADH:FMN-oxidoreductase–luciferase was investigated with addition of organic solvents (glycerol and sucrose), because bioluminescent enzyme systems based on bacterial luciferases offer a unique and general tool for analysis of the many analytes and enzymes in the environment, research, and clinical laboratories and other fields. The possibility of stabilization and increase of activity of the coupled enzyme system NADH:FMN-oxidoreductase–luciferase activity in vicious aqueous-organic mixtures have been shown.

Keywords: coupled enzyme system of bioluminescence bacteria NAD(P)H:FMN-oxidoreductase–luciferase, glycerol, stabilization of enzymes, sucrose

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Authors: Gehan A. Hegazy, Hesham N. Mustafa, Sally A. El Awdan, Marawan AbdelBaset

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Doxorubicin (DOX) is a chemotherapeutic agent used for the treatment of different cancers and its clinical usage is hindered by the oxidative injury-related cardiotoxicity. This work aims to declare if the harmful effects of DOX on the heart can be alleviated with the use of Coenzyme Q10 (CoQ10) or L-carnitine. The study was performed on seventy-two female Wistar albino rats divided into six groups, 12 animals each: Control group; DOX group (10 mg/kg); CoQ10 group (200 mg/kg); L-carnitine group (100 mg/kg); DOX + CoQ10 group; DOX + L-carnitine group. CoQ10 and L-carnitine treatment orally started five days before a single dose of 10 mg/kg DOX that injected intraperitoneally (IP) then the treatment continued for ten days. At the end of the study, serum biochemical parameters of cardiac damage, oxidative stress indices, and histopathological changes were investigated. CoQ10 or L-carnitine showed noticeable effects in improving cardiac functions evidenced reducing serum enzymes as serum interleukin-1 beta (IL-1), tumor necrosis factor alpha (TNF-), leptin, lactate dehydrogenase (LDH), Cardiotrophin-1, Troponin-I and Troponin-T. Also, alleviate oxidative stress, decrease of cardiac Malondialdehyde (MDA), Nitric oxide (NO) and restoring cardiac reduced glutathione levels to normal levels. Both corrected the cardiac alterations histologically and ultrastructurally. With visible improvements in -SMA, vimentin and eNOS immunohistochemical markers. CoQ10 or L-carnitine supplementation improves the functional and structural integrity of the myocardium.

Keywords: CoQ10, doxorubicin, L-Carnitine, cardiotoxicity

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Authors: Sonam Agarwal, Renu Bist

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Current study emphasizes the toxic effects of dichlorvos on serum in terms of oxidative stress. Meanwhile, a protective action of bacoside A and bromelain was investigated against the biochemical alterations in serum. The experimental design included six groups of mice: saline was given as a vehicle to the control mice (group I). Mice belonging to groups II, III and IV, were administered with dichlorvos (40 mg/kg b.w.), bromelain and bacoside A, respectively. The fifth group received a combination of bromelain and bacoside A. In group VI, Bacoside A, and bromelain were administered 20 minutes prior to exposure of dichlorvos. Thiobarbituric acid reactive substances (TBARS), protein carbonyl content (PCC), catalase (CAT), superoxide dismutase (SOD), glutathione peroxidase (GPx) and reduced glutathione (GSH) level were used as biochemical test of toxic action for dichlorvos intoxication. Significantly increased TBARS and PCC level in second group suggests that dichlorvos enhances the production of free radicals in serum of mice (p< 0.05). Antioxidants treatment significantly decreased the levels of TBARS and PCC (p< 0.05). Dichlorvos administration causes a significant reduction in the level of CAT, SOD, GPx and GSH (p< 0.05) which was restored significantly by co-administration of bromelain and Bacoside A in dichlorvos exposed mice (p< 0.05). Treatment of bromelain and Bacoside A in combination served as better scavengers of toxicity induced by dichlorvos.

Keywords: bacoside A, bromelain, dichlorvos, serum

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Authors: Surendra Bodakhe

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In this investigation, anticataract activity was determined using cataract formation in developing chick embryo by hydrocortisone. Lenses were evaluated firstly for the extent of opacity and secondly, for lens glutathione (GSH) levels. Betulinic acid was isolated from the chloroform fraction of the crude ethanolic extract of Bauhinia variegata bark (SBE). Fourteen days old Australorp fertilized eggs were divided into different groups of six eggs each. After 24 hrs incubation in a humidified incubator (37οC), at 15 days of age; hydrocortisone (0.25µM/0.2ml/egg) was administered to the chorioallantoic membrane of chick embryos through a small hole in the egg shell on the air sack. Ascorbic acid (standard) or Betulinic acid (test) were administered at 3, 10 and 20 hr after hydrocortisone administration at a specified dose. The puncture was sealed with a cellophane tape and eggs were incubated for 48 hrs in a humidified incubator at 37οC. After 48 hrs, the lenses were isolated for the determination of the extent of opacity and Glutathione level. The betulinic acid prevented the opacification of the chick embryo lenses induced by hydrocortisone. The betulinic acid also prevented the decline of GSH content caused by hydrocortisone. The results indicate that betulinic acid protect the cataract formation in chick embryo lenses induced by hydrocortisone.

Keywords: betulinic acid, cataract, cloudiness, ovine

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Authors: Ayse Ozge Demir, Nihat Mert

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This study has been carried out in order to determine the polymorphic traits of various biochemical parameters in goat breeds which are native to Turkey. For this purpose, Angora and Hair goats breeds were chosen as live materials. Two different herds for each breed were selected from Ankara and Antalya, respectively. Blood samples were taken from a total of 120 goats aged between 2 and 4 which was made up of 60 Angora goats and 60 Hair goats. All which derived equally from 4 lots of herds. Analyses were performed for the polymorphic determination of the Haemoglobin (Hb), Transferrine (Tf), Ceruloplasmin (Cp) and Glutathione (GSH). Hb types were determined by starch gel electrophoresis and Tf types were detected by SDS-PAGE electrophoresis. Furthermore, Cp and GSH analyses were performed by spectrophotometrically. Following the analysis, Hb types were found as 3 genotypes (AA, AB, BB) controlled by 2 allel genes. Tf types were found as 6 genotypes (AA, AB, AC, BB, BC, CC) controlled by 3 allele genes. Findings for Hb was in line with the Hardy-Weinberg Equilibrium (HWE) in Angora goats while the Hair goat was not found to be in line. Moreover, Tf was found in line with the HWE for 2 separate goat breeds. The levels of Cp and GSH of two breeds were significantly different from other (P<0.0001). The findings are recorded as a source of reference for prospective polymorphism studies.

Keywords: electrophoresis, genetic resources, goats, spectrophotometer

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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

Procedia PDF Downloads 260

Authors: Mads H. Clausen

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Plant cell walls are structurally complex and contain a larger number of diverse carbohydrate polymers. These plant fibers are a highly valuable bio-resource and the focus of food, energy and health research. We are interested in studying the interplay of plant cell wall carbohydrates with proteins such as enzymes, cell surface lectins and antibodies. However, detailed molecular level investigations of such interactions are hampered by the heterogeneity and diversity of the polymers of interest. To circumvent this, we target well-defined oligosaccharides with representative structures that can be used for characterizing protein-carbohydrate binding. The presentation will highlight chemical syntheses of plant cell wall oligosaccharides from our group and provide examples from studies of their interactions with proteins.

Keywords: oligosaccharides, carbohydrate chemistry, plant cell walls, carbohydrate-acting enzymes

Procedia PDF Downloads 285

Authors: Ayodeji Fasuyi

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Telfairia occidentalis is a leafy vegetable commonly grown in the tropics for nutritional benefits. The use of enzymes and probiotics is becoming prominent due to the ban on antibiotics as growth promoters in many parts of the world. It is conceived that with enzymes and probiotics additives, fibrous leafy vegetables can be incorporated into poultry feeds as protein source. However, certain antinutrients were also found in the leaves of Telfairia occidentalis. Four broiler starter and finisher diets were formulated for the two phases of the broiler experiments. A mixture of fiber degrading enzymes, Roxazyme G2 (combination of cellulase, glucanase and xylanase) and probiotics (Turbotox), a growth promoter, were used in broiler diets at 1:1. The Roxazyme G2/Turbotox mixtures were used in diets containing four varying levels of Telfairia occidentalis leaf meal (TOLM) at 0, 10, 20 and 30%. Diets 1 were standard broiler diets without TOLM and Roxazyme G2 and Turbotox additives. Diets 2, 3 and 4 had enzymes and probiotics additives. Certain mineral elements such as Ca, P, K, Na, Mg, Fe, Mn, Cu and Zn were found in notable quantities viz. 2.6 g/100 g, 1.2 g/100 g, 6.2 g/100 g, 5.1 g/100 g, 4.7 g/100 g, 5875 ppm, 182 ppm, 136 ppm and 1036 ppm, respectively. Phytin, phytin-P, oxalate, tannin and HCN were also found in ample quantities viz. 189.2 mg/100 g, 120.1 mg/100 g, 80.7 mg/100 g, 43.1 mg/100 g and 61.2 mg/100 g, respectively. The average weight gain was highest at 46.3 g/bird/day for birds on 10% TOLM diet but similar (P > 0.05) to 46.2 g/bird/day for birds on 20% TOLM. The feed conversion ratio (FCR) of 2.27 was the lowest and optimum for birds on 10% TOLM although similar (P > 0.05) to 2.29 obtained for birds on 20% TOLM. FCR of 2.61 was the highest at 2.61 for birds on 30% TOLM diet. The lowest FCR of 2.27 was obtained for birds on 10% TOLM diet although similar (P > 0.05) to 2.29 for birds on 20% TOLM diet. Most carcass characteristics and organ weights were similar (P > 0.05) for the experimental birds on the different diets except for kidney, gizzard and intestinal length. The values for kidney, gizzard and intestinal length were significantly higher (P < 0.05) for birds on the TOLM diets. The nitrogen retention had the highest value of 72.37 ± 0.10% for birds on 10% TOLM diet although similar (P > 0.05) to 71.54 ± 1.89 obtained for birds on the control diet without TOLM and enzymes/probiotics mixture. There was evidence of a better utilization of TOLM as a plant protein source. The carcass characteristics and organ weights all showed evidence of uniform tissue buildup and muscles development particularly in diets containing 10% of TOLM level. There was also better nitrogen utilization in birds on the 10% TOLM diet. Considering the cheap cost of TOLM, it is envisaged that its introduction into poultry feeds as a plant protein source will ultimately reduce the cost of poultry feeds.

Keywords: Telfairia occidentalis leaf meal, enzymes, probiotics, additives

Procedia PDF Downloads 116

Authors: Reshmi Vijayan

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Alkaline protease is one of the most important enzymes in the biological world. Microbial production of alkaline protease is getting more attention from researchers due to its unique properties and substantial activity. Microorganisms are the most common sources of commercial enzymes due to their physiological and biochemical properties. The study was conducted on Ayiramthenghu mangrove sediments to isolate protease producing bacteria. All the isolates were screened for proteolytic activity on a skim milk agar plate at 37˚C for 48hrs. Protease activities were determined by the formation of a clear zone around the colonies on Skim milk agar medium. The activity of the enzyme was measured by the tyrosine standard curve, and it was found to be 0.186285 U/ml/min.

Keywords: protease, protease assay, skim milk agar medium, mangrove ecosystem

Procedia PDF Downloads 74

Authors: T. H. Derdzyan, K. A. Ghazaryan, G. A. Gevorgyan

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Beta-glucosidase, chitinase, leucine-aminopeptidase, acid phosphomonoestearse and acetate-esterase enzyme activities in the soils under the impact of metallurgical industrial activity in Lori marz (district) were investigated. The results of the study showed that the activities of the investigated enzymes in the soils decreased with increasing distance from the Shamlugh copper mine, the Chochkan tailings storage facility and the ore transportation road. Statistical analysis revealed that the activities of the enzymes were positively correlated (significant) to each other according to the observation sites which indicated that enzyme activities were affected by the same anthropogenic factor. The investigations showed that the soils were polluted with heavy metals (Cu, Pb, As, Co, Ni, Zn) due to copper mining activity in this territory. The results of Pearson correlation analysis revealed a significant negative correlation between heavy metal pollution degree (Nemerow integrated pollution index) and soil enzyme activity. All of this indicated that copper mining activity in this territory causing the heavy metal pollution of the soils resulted in the inhabitation of the activities of the enzymes which are considered as biological catalysts to decompose organic materials and facilitate the cycling of nutrients.

Keywords: Armenia, metallurgical industrial activity, heavy metal pollutionl, soil enzyme activity

Procedia PDF Downloads 272

Authors: Ilma Robo, Saimir Heta, Edona Hasanaj, Vera Ostreni

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The hybrid layer, the way it is created and how it is protected against degradation over time, is the key to the clinical success of a composite restoration. The composite supports the dentinal structure exactly with the realized surface of microretension. Thus, this surface is in direct proportion to its size versus the duration of clinical use of composite dental restoration. Micro-retention occurs between dentin or acidified enamel and adhesive resin extensions versus pre-prepared spaces, such as hollow dentinal tubules. The way the adhesive resin binds to the acidified dentinal structure depends on the physical or chemical factors of this interrelationship between two structures with very different characteristics. During the acidification process, a precursor to the placement of the adhesive resin layer, activation of metaloproteinases of dental origin occurs, enzymes which are responsible for the degradation of the hybrid layer. These enzymes have expressed activity depending on the presence of Zn2 + or Ca2 + ions. There are several ways to inhibit these enzymes, and consequently, there are several ways to inhibit the degradation process of the hybrid layer. The study aims to evaluate chlorhexidine as a solution element, inhibitor of dentin activated metalloproteinases, as a result of the application of acidification. This study aims to look at this solution in advantage or contraindication theories, already published in the literature.

Keywords: hybrid layer, chlorhexidine, degradation, application

Procedia PDF Downloads 114

Authors: Njabe Christelle

Abstract:

Fungal products are essential building blocks for change towards a more sustainable future for our planet. In nature, fungi are special in breaking down plant material by means of a rich spectrum of plant cell wall degrading enzymes. Enzymes serve as catalysts in organic synthesis. Imagine the immense benefits that the known 250000 plant genes might provide in the future through scientific investigation. Plants are the primary basis for human sustenance, used directly for food, clothing, and shelter or indirectly in processed form and through animal feeding. Fungi are the only organisms known to extensively degrade lignin, a major component of wood. Although humans cannot digest cellulose and lignin, many fungi, through their assimilation of these substances, produce food in the form of edible mushrooms.

Keywords: plants, fungi, sustainable use, planet earth

Procedia PDF Downloads 56

Authors: Zakir Hossain, Arzu Pervin, Halima Jahan, Rabeya Akter, Abdel Omri

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Replacement of fish meal with soybean meal is an effective way to relieve the pressure on fish meal as the supply of this feed ingredient is dwindling and certainly is not sustainable in long term at present levels in commercial feeds. This study was designed to determine the effect of fishmeal (FM) replacement with soybean meal (SBM) in diet on growth, digestive enzyme activity, antioxidation and gut histomorphology of tilapia (Oreochromis niloticus). Five diets were formulated where SBM0 contained 100% FM, FM substituted with graded levels of a mix of SBM to replace 25% (SBM25), 50% (SBM50), 75% (SBM75) and 100% (SBM100) of FM. Juvenile tilapia having weight and length of 6.60±0.13 g and 5.42±0.17 cm were randomly divided into five treatment groups having 40 individual each group and fed to visual satiation for 90 days. Diet with SBM was increased significant in body weight gain and specific growth rate in fish compared to the fish fed with SBM100. Fish having the similar weight (74.34±5.41 g) fed the diets SBM50, SBM75 and SBM100 containing higher level of SBM showed significantly longer intestine compared to SBM0. Villus height of stomach and intestine were significantly greater in the fish fed with the diets SBM0, SBM25 and SBM50 compared to SBM100. Muscular thickness was inversely changed with the increasing villus height. Protease activity was increased significantly in stomach, anterior and posterior intestine of fish fed with SBM0 and SBM25 compared to SBM100. In anterior and posterior segment of intestine, significantly higher lipase activity was observed in fish fed with the diets SBM0 and SBM25 compared to diet SBM100. In stomach, amylase activity was also significantly greater in SBM0 compared to SBM100. The antioxidant enzymes including catalase and superoxide dismutase of liver were significantly (P < 0.05) higher in the O. niloticus fed SBM100 compared to the ones fed SBM0. These results suggest that the replacement of FM upto 75% with SBM could be possible considering the growth performances, gut health and activities digestive enzymes and antioxidant enzymes in O. niloticus.

Keywords: soybean meal, fish meal, digestive enzymes, anti-oxidant enzymes

Procedia PDF Downloads 143