Search results for: pectolytic enzymes.

Authors: Zh. B. Suleimenova, Zh. K. Rakhmetova, R. K. Blieva, A. E. Nurlybayeva

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The overall objective of this research is a strain improvement technology for efficient pectinase production. A novel cells cultivation technology by immobilization of fungal cells has been studied in long time continuous fermentations. Immobilization was achieved by using of new material for absorption of stores of immobilized cultures which was for the first time used for immobilization of microorganisms. Effects of various conditions of nitrogen and carbon nutrition on the biosynthesis of pectolytic enzymes in Aspergillus awamori 1-8 strain were studied. Proposed cultivation technology along with optimization of media components for pectinase overproduction led to increased pectinase productivity in Aspergillus awamori 1-8 from 7 to 8 times. Proposed technology can be applied successfully for production of major industrial enzymes such as α-amylase, protease, collagenase etc.

Keywords: Aspergillus awamori, immobilization, pectolytic enzymes.

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Authors: Justina I. R. Udotong

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Diagnostic enzymes like aspartate aminotransferase (AST), alanine aminotransferase (ALT) and alkaline phosphatase (ALP) were determined as indices of heavy metal pollution in Tilapia guinensis. Three different sets of fishes treated with lead (Pb), iron (Fe) and copper (Cu) were used for the study while a fourth group with no heavy metal served as a control. Fishes in each of the groups were exposed to 2.65mg/l of Pb, 0.85mg/l of Fe and 0.35 mg/l of Cu in aerated aquaria for 96 hours. Tissue fractionation of the liver tissues was carried out and the three diagnostic enzymes (AST, ALT, and ALP) were estimated. Serum levels of the same diagnostic enzymes were also measured. The mean values of the serum enzyme activity for ALP in each experimental group were 19.5±1.62, 29.67±2.17 and 1.15±0.27 IU/L for Pb, Fe and Cu groups compared with 9.99±1.34 IU/L enzyme activity in the control. This result showed that Pb and Fe caused increased release of the enzyme into the blood circulation indicating increased tissue damage while Cu caused a reduction in the serum level as compared with the level in the control group. The mean values of enzyme activity obtained in the liver were 102.14±6.12, 140.17±2.06 and 168.23±3.52 IU/L for Pb, Fe and Cu groups, respectively compared to 91.20±9.42 IU/L enzyme activity for the control group. The serum and liver AST and ALT activities obtained in Pb, Fe, Cu and control groups are reported. It was generally noted that the presence of the heavy metal caused liver tissues damage and consequent increased level of the diagnostic enzymes in the serum.

Keywords: Diagnostic enzymes, enzyme activity, heavy metals, tissues investigations.

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Authors: Parisa Farzaneh, Azade Harati

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Mushrooms or macro-fungi, as an important superfood, contain many bioactive compounds, particularly bio-peptides. In this research, mushroom proteins were extracted by buffer or buffer plus salt (0.15 M), along with ultrasound bath to extract the intercellular protein. As a result, the highest amount of proteins in mushrooms were categorized into albumin. Proteins were also hydrolyzed and changed into peptides through endogenous and exogenous proteases, including gastrointestinal enzymes. The potency of endogenous proteases was also higher in Agaricus bisporus than Terfezia claveryi, as their activity ended at 75 for 15 min. The blanching process, endogenous enzymes, the mixture of gastrointestinal enzymes (pepsin-trypsin-α-chymotrypsin or trypsin- α-chymotrypsin) produced the different antioxidant and antibacterial hydrolysates. The peptide fractions produced with different cut-off ultrafilters also had various levels of radical scavenging, lipid peroxidation inhibition, and antibacterial activities. The bio-peptides with the superior bio-activities (less than 3 kD of T. claveryi) were resistance to various environmental conditions (pH and temperatures). Therefore, they are good options to be added in nutraceutical and pharmaceutical preparations or functional foods, even during processing. 

Keywords: Bio-peptides, mushrooms, gastrointestinal enzymes, bioactivities.

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Authors: Seung Pil Pack

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Deaminated lesions were produced via nitrosative oxidation of natural nucleobases; uracul (Ura, U) from cytosine (Cyt, C), hypoxanthine (Hyp, H) from adenine (Ade, A), and xanthine (Xan, X) and oxanine (Oxa, O) from guanine (Gua, G). Such damaged nucleobases may induce mutagenic problems, so that much attentions and efforts have been poured on the revealing of their mechanisms in vivo or in vitro. In this study, we employed these deaminated lesions as useful probes for analysis of DNA-binding/recognizing proteins or enzymes. Since the pyrimidine lesions such as Hyp, Oxa and Xan are employed as analogues of guanine, their comparative uses are informative for analyzing the role of Gua in DNA sequence in DNA-protein interaction. Several DNA oligomers containing such Hyp, Oxa or Xan substituted for Gua were designed to reveal the molecular interaction between DNA and protein. From this approach, we have got useful information to understand the molecular mechanisms of the DNA-recognizing enzymes, which have not ever been observed using conventional DNA oligomer composed of just natural nucleobases.

Keywords: Deaminated lesion, DNA-protein interaction, DNA-recognizing enzymes

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Authors: Stella O. Olubodun, George E. Eriyamremu

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The study examined the effect of Bonny Light whole crude oil (WC) and its water soluble fraction (WSF) on the activities of antioxidant enzymes (catalase (CAT) and superoxide dismutase (SOD)) and crude mitochondria ATPases in the radicle of germinating bean (Vigna unguiculata). The percentage germination, level of lipid peroxidation, antioxidant enzyme and mitochondria Ca2+ and Mg2+ ATPase activities were measured in the radicle of bean after 7, 14 and 21 days post germination. Viable bean seeds were planted in soils contaminated with 10ml, 25ml and 50ml of whole crude oil (WC) and its water soluble fraction (WSF) to obtain 2, 5 and 10% v/w crude oil contamination. There was dose dependent reduction of the number of bean seeds that germinated in the contaminated soils compared with control (p<0.001). The activities of the antioxidant enzymes, as well as, adenosine triphosphatase enzymes, were also significantly (p<0.001) altered in the radicle of the plants grown in contaminated soil compared with the control. Generally, the level of lipid peroxidation was highest after 21 days post germination when compared with control. Stress to germinating bean caused by Bonny Light crude oil or its water soluble fraction resulted in adaptive changes in crude mitochondria ATPases in the radicle.

Keywords: Antioxidant enzymes, Bonny Light crude oil, Radicle, Mitochondria ATPases.

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Authors: E. Hosseini, M. Kadivar, M. Shahedi

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Malting is usually carried out on intact barley seed, while hull is still attached to it. In this study, oat grain with and without hull was subjected to controlled germination to optimize its enzymes activity, in such a way that lipase has the lowest and α- amylase and proteinase the highest activities. Since pH has a great impact on the activity of the enzymes, the pH of germination media was set up to 3 to 8. In dehulled oats, lipase and α-amylase had the lowest and highest activities in pHs 3 and 6, respectively whereas the highest proteinase activity was evidenced at pH 7 and 4 in the oats with and without hull respectively. While measurements indicated that the effect of hull on the enzyme activities particularly in lipase and amylase at each level of the pH are significantly different, the best results were obtained in those samples in which their hull had been removed. However, since the similar lipase activity in germinated dehulled oat were recorded at the pHs 4 and 5, therefore it was concluded that pH 5 in dehulled oat seed may provide the optimum enzyme activity for all the enzymes.

Keywords: Enzyme activity, malting, oat, optimization.

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Authors: D. Scalone, R. Palmeri, F. Licciardello, G. Muratore, A. Todaro, G. Spagna

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Prickly pear fruit (Opuntia ficus indica L. Miller) belongs to the Cactaceae family. This species is very sensitive to low storage temperatures (< 5°C) which cause damages. The fruits can be peeled, suitably packaged and successfully commercialized as a ready-to-eat product. The main limit to the extension of the shelf life is the production of off-flavors due to different factors, the growth of microorganisms and the action of endogenous enzymes. Lipoxygenase (LOX) and Pectinesterase (PE) are involved in fruit degradation. In particular, LOX pathway is directly responsible for lipid oxidation, and the subsequent production of off-flavours, while PE causes the softening of fruit during maturation. They act on the texture and shelf-life of post-harvest, packaged fruits, as a function of the the grown of microorganisms and packaging technologies used. The aim of this work is to compare the effect of different packaging technologies on the shelf life extension of ready-to-eat prickly pear fruits with regards for the enzymes activities.

Keywords: Enzymes, packaging, prickly pear, shelf life.

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Authors: Debora B. Moretti, Wiolene M. Nordi, Thaline Maira P. Cruz, José Eurico P. Cyrino, Raul Machado-Neto

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Enzyme activity was evaluated in the intestine of juvenile dourado (Salminus brasiliensis) fed with diets containing 0, 10 or 20% of lyophilized bovine colostrum (LBC) inclusion for either 30 or 60 days. The intestinal enzymes acid and alkaline phosphatase (ACP and ALP, respectively), non-specific esterase (NSE), lipase (LIP), dipeptidyl aminopeptidase IV (DAP IV) and leucine aminopeptidase (LAP) were studied using histochemistry in four intestinal segments (S1, S2, S3 and posterior intestine). Weak proteolitic activity was observed in all intestinal segments for DAP IV and LAP. The activity of NSE and LIP was also weak in all intestines, except for the moderate activity of NSE in the S2 of 20% LBC group after 30 days and in the S1 of 0% LBC group after 60 days. The ACP was detected only in the S2 and S3 of the 10% LBC group after 30 days. Moderate and strong staining was observed in the first three intestinal segments for ALP and weak activity in the posterior intestine. The activity of DAP IV, LAP and ALP were also present in the cytoplasm of the enterocytes. In the present results, bovine colostrum feeding did not cause alterations in activity of intestinal enzymes.

Keywords: Carnivorous fish, enterocyte, intestinal epithelium, teleost.

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Authors: Niraj A. Ghanwate, P V Thakare, P R Bhise, Ashish Dhanke, Shubhangi Apotikar

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Urinary Tract Infections (UTI) account for an estimated 25-40% nosocomial infection, out of which 90% are associated with urinary catheter, called Catheter associated urinary tract infection (CAUTI). The microbial populations within CAUTI frequently develop as biofilms. In the present study, microbial contamination of indwelling urinary catheters was investigated. Biofilm forming ability of the isolates was determined by tissue culture plate method. Prevention of biofilm formation in the urinary catheter by Pseudomonas aeruginosa was also determined by coating the catheter with some enzymes, gentamycin and EDTA. It was found that 64% of the urinary catheters get contaminated during the course of catheterization. Of the total 6 isolates, biofilm formation was seen in 100% Pseudomonas aeruginosa and E. coli, 90% in Enterococci, 80% in Klebsiella and 66% in S. aureus. It was noted that the biofilm production by Pseudomonas was prolonged by 7 days in amylase, 8 days in protease, 6 days in lysozyme, 7days in gentamycin and 5 days in EDTA treated catheter.

Keywords: CAUTI, biofilm, enzymes, EDTA, Pseudomonas.

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Authors: Sérgio F. Sousa, Nuno M. F. S. A. Cerqueira, Marta A. S. Perez, Irina S. Moreira, António J. M.Ribeiro, Ana R. A. P. Neves, Maria J. Ramos, Pedro A. Fernandes

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In this work we report the recent progresses that have been achieved by our group in the last half decade on the field of computational proteomics. Specifically, we discuss the application of Molecular Dynamics Simulations and Electronic Structure Calculations in drug design, in the clarification of the structural and dynamic properties of proteins and enzymes and in the understanding of the catalytic and inhibition mechanism of cancer-related enzymes. A set of examples illustrate the concepts and help to introduce the reader into this important and fast moving field.

Keywords: Enzyme, Molecular Dynamics, Protein, Quantum Mechanics.

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Authors: Simon Brown, Noorzaid Muhamad, David C Simcock

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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: A. Tomruk, G. Guler, B. Tandogan, E. Ozgur, N.N. Ulusu, N. Seyhan

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In the present study, we aimed to design the intrauterine and extrauterine exposure to 1800 MHz GSM-like RF radiation and investigate its possible bio-effects on infant female rabbits. Totally thirty-six New Zealand White female rabbits, onemonth old, were randomly divided into four groups which are composed of 9 rabbits; i. Group I [Intrauterine (IU) exposure(-); Extrauterine (EU) exposure (-)], Group II [IU exposure (-); EU exposure (+)], Group III [IU exposure(+);EU exposure(-)], Group IV [IU exposure (+);EU exposure(+)]. The master regulatory enzymes activities of pentose phosphate pathway (glucose-6-phosphate dehydrogenase, G-6PD; 6-phosphogluconate dehydrogenase, 6- PGDH) and glutathione-dependent metabolism (glutathione peroxidase, GSH-Px; glutathione reductase, GR; glutathione Stransferase, GST, thioredoxin reductase, TRx) were analyzed in liver tissues of young female rabbits. Decreased G-6PD, 6-PGD, GSH-Px, GR activities were found in Group III compared to Group I (p<0.05, Mann Whitney). Increased GSH-px and TRx activities were found in Group IV compared to Group I (p<0.05, Mann Whitney). It can be concluded that the intrauterine and extrauterine exposure to GSMlike RF radiation may influence the liver regulatory enzymes activities.

Keywords: Radiofrequency (RF), intrauterine (IU) andextrauterine (EU) exposure, infant female rabbits.

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Authors: A. Maitarad, P. Maitarad

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In order to find the particular interaction energy between cylcloguanil and the amino acids surrounding the pocket of wild type and quadruple mutant type PfDHFR enzymes, the MP2 method with basis set 6-31G(d,p) level of calculations was performed. The obtained interaction energies found that Asp54 has the strongest interaction energy to both wild type and mutant type of - 12.439 and -11.250 kcal/mol, respectively and three amino acids; Asp54, Ile164 and Ile14 formed the H-bonding with cycloguanil drug. Importantly, the mutation at Ser108Asn was the key important of cycloguanil resistant with showing repulsive interaction energy.

Keywords: Cycloguanil, DHFR, malaria disease, interactionenergy, quantum calculations

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Authors: Tudor Petreuş, Andrei Neamţu, Cristina Dascălu, Paul Dan Sîrbu, Carmen E. Cotrutz

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Matrix metalloproteinases (MMP) are a class of structural and functional related enzymes involved in altering the natural elements of the extracellular matrix. Most of the MMP structures are cristalographycally determined and published in WorldWide ProteinDataBank, isolated, in full structure or bound to natural or synthetic inhibitors. This study proposes an algorithm to replace missing crystallographic structures in PDB database. We have compared the results of a chosen docking algorithm with a known crystallographic structure in order to validate enzyme sites reconstruction there where crystallographic data are missing.

Keywords: matrix metalloproteinases, molecular docking, structure superposition, surface complementarity.

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Authors: Nasim Shaibani, Saba Ghazvini, Mohammad R. Andalibi, Soheila Yaghmaei

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Nowadays there is a growing interest in biofuel production in most countries because of the increasing concerns about hydrocarbon fuel shortage and global climate changes, also for enhancing agricultural economy and producing local needs for transportation fuel. Ethanol can be produced from biomass by the hydrolysis and sugar fermentation processes. In this study ethanol was produced without using expensive commercial enzymes from sugarcane bagasse. Alkali pretreatment was used to prepare biomass before enzymatic hydrolysis. The comparison between NaOH, KOH and Ca(OH)2 shows NaOH is more effective on bagasse. The required enzymes for biomass hydrolysis were produced from sugarcane solid state fermentation via two fungi: Trichoderma longibrachiatum and Aspergillus niger. The results show that the produced enzyme solution via A. niger has functioned better than T. longibrachiatum. Ethanol was produced by simultaneous saccharification and fermentation (SSF) with crude enzyme solution from T. longibrachiatum and Saccharomyces cerevisiae yeast. To evaluate this procedure, SSF of pretreated bagasse was also done using Celluclast 1.5L by Novozymes. The yield of ethanol production by commercial enzyme and produced enzyme solution via T. longibrachiatum was 81% and 50% respectively.

Keywords: Alkali pretreatment, bioethanol, cellulase, simultaneous saccharification and fermentation, solid statefermentation, sugarcane bagasse

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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, a significant amount 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 work group. 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: 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: 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.

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

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

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The hybrid layer (HL), 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 micro-retention. 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 metalloproteinases of dental origin occurs, enzymes which are responsible for the degradation of the HL. 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 HL. The study aim is to evaluate chlorhexidine (CHX) 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, smear layer.

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

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Beta-glucosidase, chitinase, leucine-aminopeptidase, acid phosphomonoesterase 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.

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Authors: Nirmal J.I. Kumar, Anubhuti A. Bora, Manmeet K. Amb

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Application of pesticides in the paddy fields has deleterious effects on non-target organisms including cyanobacteria which are photosynthesizing and nitrogen fixing micro-organisms contributing significantly towards soil fertility and crop yield. Pesticide contamination in the paddy fields has manifested into a serious global environmental concern. To study the effect of one such pesticide, three cyanobacterial strains; Anabaena fertilissima, Aulosira fertilissima and Westiellopsis prolifica were selected for their stress responses to an Organochlorine insecticide - 6, 7, 8, 9, 10, 10-hexachloro-1, 5, 5a, 6, 9, 9a-hexahydro-6, 9-methano-2, 4, 3- benzodioxathiepine-3-oxide, with reference to their photosynthesic pigments-chlorophyll-a and carotenoids as well as accessory pigments-phycobiliproteins (phycocyanin, allophycocyanin and phycoerythrin), stress induced biochemical metabolites like carbohydrates, proteins, amino acids, phenols and enzymes-nitrate reductase, glutamine synthetase and succinate dehydrogenase. All the three cyanobacterial strains were adversely affected by the insecticide doses and inhibition was dose dependent. Reduction in photosynthetic and accessory pigments, metabolites, nitrogen fixing and respiratory enzymes of the test organisms were accompanied with an initial increase in their total protein at lower Organochlorine doses. On the other hand, increased amount of phenols in all the insecticide treated concentrations was indicative of stressed activities of the organisms.

Keywords: biochemical metabolites, endosulfan, enzymes, pigments

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Authors: Ayuko Itsuki, Sachiyo Aburatani

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Soil enzyme activities in Kasuga-yama Hill Primeval Forest (Nara, Japan) were examined to determine levels of mineralization and metabolism. Samples were selected from the soil surrounding laurel-leaved (B_B-1) and Carpinus japonica (B_B-2 and P_w) trees for analysis. Cellulase, β-xylosidase, and protease activities were higher in B_B-1 samples those in B_B-2 samples. These activity levels corresponded to the distribution of cellulose and hemicellulose in the soil horizons. Cellulase, β-xylosidase, and chymotrypsin activities were higher in soil from the P_w forest than in that from the B_B-2 forest. The relationships between the soil enzymes calculated by Spearman’s rank correlation indicate that the interactions between enzymes in B_B-2 samples were more complex than those in P_w samples.

Keywords: Comparative analysis, enzyme activities, forest soil, Spearman’s rank correlation.

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Authors: K. Mahmoud, T. Shalahmetova, Sh. Deraz

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The objective of the present study was to examine the dose-response relationships between antioxidant parameters and liver contaminant levels of Kazakhstan light crude oil (KLCO) in albino rats. The animals were repeatedly exposed, by intraperitoneal injection, to low dosages (0.5–1.5 ml/kg) of KLCO. Rats exposed to these doses levels did not show any apparent symptoms of intoxication. Serum aminotransferases increased significantly (p<0.05) while after 8-d recovered to control level. hepatic superoxide dismutase (SOD) activity significantly increased after 1 and 3-d except with 0.5 ml while with increased exposure time it deceased insignificantly after 5-d and significantly after 8-d. Dose and time-dependent increases were observed in the levels of hepatic glutathione-S-transeferase (GST) and conjugated diene (CD) while malondialdehyde (MDA) was evolved after 8 with the dose of 1.5 ml. the data obtained indicate impaired liver function in the rats intoxicated with KLCO as manifested by elevated liver enzymes.

Keywords: Crude oil; superoxide dismutase, glutathione-Stranseferase, malondialdehyde, conjugated diene, aminotransferases.

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Authors: Yahia Massinissa, Kalla A, Yahia M, Benbia S

Abstract:

A lot of recent research have spoken on the relation between the increase of the homocysteinemia and some kinds of cancer . For that, our study was based on the research of a possible relation between the increase of the concentration of this amino-acid in the plasma and the appearance of the disease of the Acute Lymphoblastic Leukaemia in a part of Algerian children with Berber origin in the East of Algeria . The study has done on 47 ill persons with an average age of (09±06 ) years , with whom the disease has diagnosed by blood and marrow examination in the hospital of blood diseases in the CHU of Batna, and on 194 healthy witnesses of the same age. The two groups were benefited by a dosage of the concentration of the homocysteine vitamin B9 ,vitamin B12 , and also of the study of special polymorphisms of indispensable enzymes in the metabolism of this acid , and that by the use of the method ( Light cycler ) Real time PCR , on the following enzymes : MS ( C2756G ), MSR ( A66G ) ,MTHFR1 ( C677T ) and MTHFR2 (A1298C). The obtained results have revealed that the rate of the homozygote muted genotype is the less frequent in the two groups , and that exist at list one genotype of each enzyme in the ill group and in which the percentage exceed with remarkable way the same genotype in the healthy group and we notice specially the muted genotype GG of -the methionine synthetase-and the form TT of the enzyme – methyline tetra hydrofolate reductase – We notice the existence of considerable number of genotypes in the ill group lied with characteristic increase of this Amino-acid ,and that for the reduction of the biologic activity of these enzymes which become inefficient in the transfer of the homocysteine into the methionine and cause the diminution of the biologic activity of these enzymes and with consequence the reduction of the percentage of methylic radicals in the DNA of studied genes and that lead to the increase of the activity and the capacity of transcription , and it-s so probably that this last one is one of the factors of this disease especially if we know that the specific check-up of vitamins is normal and similar in the two groups , which ovoid the hypothesis of the reduction of vitamins . We notice also that the heterozygote genotype is the less in the sick category except the MTHFR2. Wild genotype is more frequent in the witness group except MSR. Even these results are partials; they open a new way in the genetic diagnosis of this malicious disease which allow a precocious diagnosis and the use of an effective and appropriated treatment in the same time.

Keywords: Genetic polymorphism, Acute Lymphoblastic Leukaemia, Biomarkers, Metabolism of homocystein

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Authors: Deepak Mohan, Sushma Sharma

Abstract:

Non-Steroidal Anti-Inflammatory Drugs (NSAIDs) are a group of widely used drugs for the treatment of rheumatoid diseases and to relieve pain and inflammation due to their analgesic anti-pyretic and anti-inflammatory properties. The therapeutic and many of the toxic effects of NSAIDs result from reversible inhibition of enzymes in the cyclooxygenase (COX) group. In the present investigation the effect of the drug on the concentration of lipids, and on the activity of the enzymes i.e. acid and alkaline phosphatase, GOT, GPT and lipid peroxidase were studied. There was a significant enhancement in the activities of both acid and alkaline phosphatase after 21 days of treatment. Proportionate increase in the MDA contents was observed after different days of diclofenac treatment. Cellular damage in the liver resulted in decrease in the activity of both GOT (Glutamate oxaloacetate transaminase) and GPT (Glutamate pyruvate transaminase) in both low and high dose groups. Significant decrease in the liver contents was also observed in both dose groups.

Keywords: Anti-inflammatory, cyclooxygenase, glutamate oxaloacetate transaminase, malondialdehyde.

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Authors: Sudarat Jiamyangyuen, Tipawan Thongsook, Riantong Singanusong, Chanida Saengtubtim

Abstract:

Rice bran is normally used as a raw material for rice bran oil production or sold as feed with a low price. Conventionally, the protein in defatted rice bran was extracted using alkaline extraction and acid precipitation, which involves in chemical usage and lowering some nutritious component. This study was conducted in order to extract of rice bran protein concentrate (RBPC) from defatted rice bran using enzymes and employing polysaccharides in a precipitating step. The properties of RBPC obtained will be compared to those of a control sample extracted using a conventional method. The results showed that extraction of protein from rice bran using enzymes exhibited the higher protein recovery compared to that extraction with alkaline. The extraction conditions using alcalase 2% (v/w) at 50 C, pH 9.5 gave the highest protein (2.44%) and yield (32.09%) in extracted solution compared to other enzymes. Rice bran protein concentrate powder prepared by a precipitation step using alginate (protein in solution: alginate 1:0.016) exhibited the highest protein (27.55%) and yield (6.84%). Precipitation using alginate was better than that of acid. RBPC extracted with alkaline (ALK) or enzyme alcalase (ALC), then precipitated with alginate (AL) (samples RBP-ALK-AL and RBP-ALC-AL) yielded the precipitation rate of 75% and 91.30%, respectively. Therefore, protein precipitation using alginate was then selected. Amino acid profile of control sample, and sample precipitated with alginate, as compared to casein and soy protein isolated, showed that control sample showed the highest content among all sample. Functional property study of RBP showed that the highest nitrogen solubility occurred in pH 8-10. There was no statically significant between emulsion capacity and emulsion stability of control and sample precipitated by alginate. However, control sample showed a higher of foaming capacity and foaming stability compared to those of sample precipitated with alginate. The finding was successful in terms of minimizing chemicals used in extraction and precipitation steps in preparation of rice bran protein concentrate. This research involves in a production of value-added product in which the double amount of protein (28%) compared to original amount (14%) contained in rice bran could be beneficial in terms of adding to food products e.g. healthy drink with high protein and fiber. In addition, the basic knowledge of functional property of rice bran protein concentrate was obtained, which can be used to appropriately select the application of this value-added product from rice bran.

Keywords: Alginate, carrageenan, rice bran, rice bran protein.

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

Abstract:

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: S. Benbia, A.Kalla, M. Yahia, K. Belhadi, A. Zidani

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Forty-five dairy cows were used to compare the enzyme activity of alkaline phosphatase (ALP), lactate dehydrogenase (LDH), α -amylase in the cervical mucus of cows during spontaneous and induced estrus using progestagen or PGF2 α and to determine whether these enzymes affect the fertility in cows with induced estrus, at the time of Al. The animals were assigned to 3 groups (no treatment, a Crestar® for 12 days, a double im injection of PGF2 α). The cows were artificially inseminated (AI). Cervical mucus samples were collected from all cows 3 to 5 min before the AI. The results are summarized as follows: ALP and α -amylase activity for spontaneous estrus were similar to those for induced estrus (P>0.05) . LDH activity levels during spontaneous and PGF2 α induced estrus was significantly lower (P < 0.001) than that in progestagene induced estrus groups. While no difference was found between the first and the third groups. Our result showed a significant difference in LDH activity levels between cows conceived with 2 or more AI and those conceived with 1 AI. The result of this study showed that the enzyme activity in cervical mucus is helpful for detection of ovulation and time of AI.

Keywords: cervical mucus, dairy cow, enzyme, induced, estrus, ovulation, AI

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Authors: P. Pasomboon, P. Chumnanpuen, T. E-kobon

Abstract:

Hyaluronic acid (HA) consists of linear heteropolysaccharides repeat of D-glucuronic acid and N-acetyl-D-glucosamine. HA has various useful properties to maintain skin elasticity and moisture, reduce inflammation, and lubricate the movement of various body parts without causing immunogenic allergy. HA can be found in several animal tissues as well as in the capsule component of some bacteria including Pasteurella multocida. This study aimed to modify a genome-scale metabolic model of Escherichia coli using computational simulation and flux analysis methods to predict HA productivity under different carbon sources and nitrogen supplement by the addition of two enzymes (GLMU2 and HYAD) from P. multocida to improve the HA production under the specified amount of carbon sources and nitrogen supplements. Result revealed that threonine and aspartate supplement raised the HA production by 12.186%. Our analyses proposed the genome-scale metabolic model is useful for improving the HA production and narrows the number of conditions to be tested further.

Keywords: Pasteurella multocida, Escherichia coli, hyaluronic acid, genome-scale metabolic model, bioinformatics.

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