Search results for: Enzymes
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 101

Search results for: Enzymes

71 Jamun Juice Extraction Using Commercial Enzymes and Optimization of the Treatment with the Help of Physicochemical, Nutritional and Sensory Properties

Authors: Payel Ghosh, Rama Chandra Pradhan, Sabyasachi Mishra

Abstract:

Jamun (Syzygium cuminii L.) is one of the important indigenous minor fruit with high medicinal value. The jamun cultivation is unorganized and there is huge loss of this fruit every year. The perishable nature of the fruit makes its postharvest management further difficult. Due to the strong cell wall structure of pectin-protein bonds and hard seeds, extraction of juice becomes difficult. Enzymatic treatment has been commercially used for improvement of juice quality with high yield. The objective of the study was to optimize the best treatment method for juice extraction. Enzymes (Pectinase and Tannase) from different stains had been used and for each enzyme, best result obtained by using response surface methodology. Optimization had been done on the basis of physicochemical property, nutritional property, sensory quality and cost estimation. According to quality aspect, cost analysis and sensory evaluation, the optimizing enzymatic treatment was obtained by Pectinase from Aspergillus aculeatus strain. The optimum condition for the treatment was 44 oC with 80 minute with a concentration of 0.05% (w/w). At these conditions, 75% of yield with turbidity of 32.21NTU, clarity of 74.39%T, polyphenol content of 115.31 mg GAE/g, protein content of 102.43 mg/g have been obtained with a significant difference in overall acceptability.

Keywords: Jamun, enzymatic treatment, physicochemical property, sensory analysis, optimization.

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70 Isolation and Screening of Fungal Strains for β-Galactosidase Production

Authors: Parmjit S. Panesar, Rupinder Kaur, Ram S. Singh

Abstract:

Enzymes are the biocatalysts which catalyze the biochemical processes and thus have a wide variety of applications in the industrial sector. β-Galactosidase (E.C. 3.2.1.23) also known as lactase, is one of the prime enzymes, which has significant potential in the dairy and food processing industries. It has the capability to catalyze both the hydrolytic reaction for the production of lactose hydrolyzed milk and transgalactosylation reaction for the synthesis of prebiotics such as lactulose and galactooligosaccharides. These prebiotics have various nutritional and technological benefits. Although, the enzyme is naturally present in almonds, peaches, apricots and other variety of fruits and animals, the extraction of enzyme from these sources increases the cost of enzyme. Therefore, focus has been shifted towards the production of low cost enzyme from the microorganisms such as bacteria, yeast and fungi. As compared to yeast and bacteria, fungal β-galactosidase is generally preferred as being extracellular and thermostable in nature. Keeping the above in view, the present study was carried out for the isolation of the β-galactosidase producing fungal strain from the food as well as the agricultural wastes. A total of more than 100 fungal cultures were examined for their potential in enzyme production. All the fungal strains were screened using X-gal and IPTG as inducers in the modified Czapek Dox Agar medium. Among the various isolated fungal strains, the strain exhibiting the highest enzyme activity was chosen for further phenotypic and genotypic characterization. The strain was identified as Rhizomucor pusillus on the basis of 5.8s RNA gene sequencing data.

Keywords: β-galactosidase, enzyme, fungus, isolation.

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69 Higher Plants Ability to Assimilate Explosives

Authors: G. Khatisashvili, M. Gordeziani, G. Adamia, E. Kvesitadze, T. Sadunishvili, G. Kvesitadze

Abstract:

The ability of agricultural and decorative plants to absorb and detoxify TNT and RDX has been studied. All tested 8 plants, grown hydroponically, were able to absorb these explosives from water solutions: Alfalfa > Soybean > Chickpea> Chikling vetch >Ryegrass > Mung bean> China bean > Maize. Differently from TNT, RDX did not exhibit negative influence on seed germination and plant growth. Moreover, some plants, exposed to RDX containing solution were increased in their biomass by 20%. Study of the fate of absorbed [1-14ðí]-TNT revealed the label distribution in low and high-molecular mass compounds, both in roots and above ground parts of plants, prevailing in the later. Content of 14ðí in lowmolecular compounds in plant roots are much higher than in above ground parts. On the contrary, high-molecular compounds are more intensively labeled in aboveground parts of soybean. Most part (up to 70%) of metabolites of TNT, formed either by enzymatic reduction or oxidation, is found in high molecular insoluble conjugates. Activation of enzymes, responsible for reduction, oxidation and conjugation of TNT, such as nitroreductase, peroxidase, phenoloxidase and glutathione S-transferase has been demonstrated. Among these enzymes, only nitroreductase was shown to be induced in alfalfa, exposed to RDX. The increase in malate dehydrogenase activities in plants, exposed to both explosives, indicates intensification of Tricarboxylic Acid Cycle, that generates reduced equivalents of NAD(P)H, necessary for functioning of the nitroreductase. The hypothetic scheme of TNT metabolism in plants is proposed.

Keywords: Higher plants, TNT, RDX, transformation.

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68 Utilization of Whey for the Production of β-Galactosidase Using Yeast and Fungal Culture

Authors: Rupinder Kaur, Parmjit S. Panesar, Ram S. Singh

Abstract:

Whey is the lactose rich by-product of the dairy industry, having good amount of nutrient reservoir. Most abundant nutrients are lactose, soluble proteins, lipids and mineral salts. Disposing of whey by most of milk plants which do not have proper pre-treatment system is the major issue. As a result of which, there can be significant loss of potential food and energy source. Thus, whey has been explored as the substrate for the synthesis of different value added products such as enzymes. β-galactosidase is one of the important enzymes and has become the major focus of research due to its ability to catalyze both hydrolytic as well as transgalactosylation reaction simultaneously. The enzyme is widely used in dairy industry as it catalyzes the transformation of lactose to glucose and galactose, making it suitable for the lactose intolerant people. The enzyme is intracellular in both bacteria and yeast, whereas for molds, it has an extracellular location. The present work was carried to utilize the whey for the production of β-galactosidase enzyme using both yeast and fungal cultures. The yeast isolate Kluyveromyces marxianus WIG2 and various fungal strains have been used in the present study. Different disruption techniques have also been investigated for the extraction of the enzyme produced intracellularly from yeast cells. Among the different methods tested for the disruption of yeast cells, SDS-chloroform showed the maximum β-galactosidase activity. In case of the tested fungal cultures, Aureobasidium pullulans NCIM 1050 was observed to be the maximum extracellular enzyme producer.

Keywords: β-galactosidase, fungus, yeast, whey.

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67 Analysis of a Lignocellulose Degrading Microbial Consortium to Enhance the Anaerobic Digestion of Rice Straws

Authors: Supanun Kangrang, Kraipat Cheenkachorn, Kittiphong Rattanaporn, Malinee Sriariyanun

Abstract:

Rice straw is lignocellulosic biomass which can be utilized as substrate for the biogas production. However, due to the property and composition of rice straw, it is difficult to be degraded by hydrolysis enzymes. One of the pretreatment methods that modify such properties of lignocellulosic biomass is the application of lignocellulose-degrading microbial consortia. The aim of this study is to investigate the effect of microbial consortia to enhance biogas production. To select the high efficient consortium, cellulase enzymes were extracted and their activities were analyzed. The results suggested that microbial consortium culture obtained from cattle manure is the best candidate compared to decomposed wood and horse manure. A microbial consortium isolated from cattle manure was then mixed with anaerobic sludge and used as inoculum for biogas production. The optimal conditions for biogas production were investigated using response surface methodology (RSM). The tested parameters were the ratio of amount of microbial consortium isolated and amount of anaerobic sludge (MI:AS), substrate to inoculum ratio (S:I) and temperature. Here, the value of the regression coefficient R2 = 0.7661 could be explained by the model which is high to advocate the significance of the model. The highest cumulative biogas yield was 104.6 ml/g-rice straw at optimum ratio of MI:AS, ratio of S:I, and temperature of 2.5:1, 15:1 and 44°C respectively.

Keywords: Lignocellulolytic biomass, microbial consortium, cellulase, biogas, Response Surface Methodology.

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66 Investigations of Metals and Metal-Antibrowning Agents Effects on Polyphenol Oxidase Activity from Red Poppy Leaf

Authors: G. Arabaci

Abstract:

Heavy metals are one of the major groups of contaminants in the environment and many of them are toxic even at very low concentration in plants and animals. However, some metals play important roles in the biological function of many enzymes in living organisms. Metals such as zinc, iron, and cooper are important for survival and activity of enzymes in plants, however heavy metals can inhibit enzyme which is responsible for defense system of plants. Polyphenol oxidase (PPO) is a copper-containing metalloenzyme which is responsible for enzymatic browning reaction of plants. Enzymatic browning is a major problem for the handling of vegetables and fruits in food industry. It can be increased and effected with many different futures such as metals in the nature and ground. In the present work, PPO was isolated and characterized from green leaves of red poppy plant (Papaverr hoeas). Then, the effect of some known antibrowning agents which can form complexes with metals and metals were investigated on the red poppy PPO activity. The results showed that glutathione was the most potent inhibitory effect on PPO activity. Cu(II) and Fe(II) metals increased the enzyme activities however, Sn(II) had the maximum inhibitory effect and Zn(II) and Pb(II) had no significant effect on the enzyme activity. In order to reduce the effect of heavy metals, the effects of metal-antibrowning agent complexes on the PPO activity were determined. EDTA and metal complexes had no significant effect on the enzyme. L-ascorbic acid and metal complexes decreased but L-ascorbic acid-Cu(II)-complex had no effect. Glutathione–metal complexes had the best inhibitory effect on Red poppy leaf PPO activity.

Keywords: Inhibition, metal, red poppy, Polyphenol oxidase (PPO).

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65 Biodegradation of Malathion by Acinetobacter baumannii Strain AFA Isolated from Domestic Sewage in Egypt

Authors: Ahmed F. Azmy , Amal E. Saafan, Tamer M. Essam, Magdy A. Amin, Shaban H. Ahmed

Abstract:

Bacterial strains capable of degradation of malathion from the domestic sewage were isolated by an enrichment culture technique. Three bacterial strains were screened and identified as Acinetobacter baumannii (AFA), Pseudomonas aeruginosa (PS1), and Pseudomonas mendocina (PS2) based on morphological, biochemical identification and 16S rRNA sequence analysis. Acinetobacter baumannii AFA was the most efficient malathion degrading bacterium, so used for further biodegradation study. AFA was able to grow in mineral salt medium (MSM) supplemented with malathion (100 mg/l) as a sole carbon source, and within 14 days, 84% of the initial dose was degraded by the isolate measured by high performance liquid chromatography. Strain AFA could also degrade other organophosphorus compounds including diazinon, chlorpyrifos and fenitrothion. The effect of different culture conditions on the degradation of malathion like inoculum density, other carbon or nitrogen sources, temperature and shaking were examined. Degradation of malathion and bacterial cell growth were accelerated when culture media were supplemented with yeast extract, glucose and citrate. The optimum conditions for malathion degradation by strain AFA were; an inoculum density of 1.5x 10^12CFU/ml at 30°C with shaking. A specific polymerase chain reaction primers were designed manually using multiple sequence alignment of the corresponding carboxylesterase enzymes of Acinetobacter species. Sequencing result of amplified PCR product and phylogenetic analysis showed low degree of homology with the other carboxylesterase enzymes of Acinetobacter strains, so we suggested that this enzyme is a novel esterase enzyme. Isolated bacterial strains may have potential role for use in bioremediation of malathion contaminated.

Keywords: Acinetobacter baumannii, biodegradation, Malathion, organophosphate pesticides.

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64 Forensic Medical Capacities of Research of Saliva Stains on Physical Evidence after Washing

Authors: Saule Mussabekova

Abstract:

Recent advances in genetics have allowed increasing acutely the capacities of the formation of reliable evidence in conducting forensic examinations. Thus, traces of biological origin are important sources of information about a crime. Currently, around the world, sexual offenses have increased, and among them are those in which the criminals use various detergents to remove traces of their crime. A feature of modern synthetic detergents is the presence of biological additives - enzymes. Enzymes purposefully destroy stains of biological origin. To study the nature and extent of the impact of modern washing powders on saliva stains on the physical evidence, specially prepared test specimens of different types of tissues to which saliva was applied have been examined. Materials and Methods: Washing machines of famous manufacturers of household appliances have been used with different production characteristics and advertised brands of washing powder for test washing. Over 3,500 experimental samples were tested. After washing, the traces of saliva were identified using modern research methods of forensic medicine. Results: The influence was tested and the dependence of the use of different washing programs, types of washing machines and washing powders in the process of establishing saliva trace and identify of the stains on the physical evidence while washing was revealed. The results of experimental and practical expert studies have shown that in most cases it is not possible to draw the conclusions in the identification of saliva traces on physical evidence after washing. This is a consequence of the effect of biological additives and other additional factors on traces of saliva during washing. Conclusions: On the basis of the results of the study, the feasibility of saliva traces of the stains on physical evidence after washing is established. The use of modern molecular genetic methods makes it possible to partially solve the problems arising in the study of unlaundered evidence. Additional study of physical evidence after washing facilitates detection and investigation of sexual offenses against women and children.

Keywords: Saliva research, modern synthetic detergents, laundry detergents, forensic medicine.

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63 Systematic Identification and Quantification of Substrate Specificity Determinants in Human Protein Kinases

Authors: Manuel A. Alonso-Tarajano, Roberto Mosca, Patrick Aloy

Abstract:

Protein kinases participate in a myriad of cellular processes of major biomedical interest. The in vivo substrate specificity of these enzymes is a process determined by several factors, and despite several years of research on the topic, is still far from being totally understood. In the present work, we have quantified the contributions to the kinase substrate specificity of i) the phosphorylation sites and their surrounding residues in the sequence and of ii) the association of kinases to adaptor or scaffold proteins. We have used position-specific scoring matrices (PSSMs), to represent the stretches of sequences phosphorylated by 93 families of kinases. We have found negative correlations between the number of sequences from which a PSSM is generated and the statistical significance and the performance of that PSSM. Using a subset of 22 statistically significant PSSMs, we have identified specificity determinant residues (SDRs) for 86% of the corresponding kinase families. Our results suggest that different SDRs can function as positive or negative elements of substrate recognition by the different families of kinases. Additionally, we have found that human proteins with known function as adaptors or scaffolds (kAS) tend to interact with a significantly large fraction of the substrates of the kinases to which they associate. Based on this characteristic we have identified a set of 279 potential adaptors/scaffolds (pAS) for human kinases, which is enriched in Pfam domains and functional terms tightly related to the proposed function. Moreover, our results show that for 74.6% of the kinase–pAS association found, the pAS colocalize with the substrates of the kinases they are associated to. Finally, we have found evidence suggesting that the association of kinases to adaptors and scaffolds, may contribute significantly to diminish the in vivo substrate crossed-specificity of protein kinases. In general, our results indicate the relevance of several SDRs for both the positive and negative selection of phosphorylation sites by kinase families and also suggest that the association of kinases to pAS proteins may be an important factor for the localization of the enzymes with their set of substrates.

Keywords: Kinase, phosphorylation, substrate specificity, adaptors, scaffolds, cellular colocalization.

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62 A Neural Computing-Based Approach for the Early Detection of Hepatocellular Carcinoma

Authors: Marina Gorunescu, Florin Gorunescu, Kenneth Revett

Abstract:

Hepatocellular carcinoma, also called hepatoma, most commonly appears in a patient with chronic viral hepatitis. In patients with a higher suspicion of HCC, such as small or subtle rising of serum enzymes levels, the best method of diagnosis involves a CT scan of the abdomen, but only at high cost. The aim of this study was to increase the ability of the physician to early detect HCC, using a probabilistic neural network-based approach, in order to save time and hospital resources.

Keywords: Early HCC diagnosis, probabilistic neural network.

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61 Influence of Hydrocarbons on Plant Cell Ultrastructure and Main Metabolic Enzymes

Authors: T. Sadunishvili, E. Kvesitadze, M. Betsiashvili, N. Kuprava, G. Zaalishvili, G. Kvesitadze

Abstract:

Influence of octane and benzene on plant cell ultrastructure and enzymes of basic metabolism, such as nitrogen assimilation and energy generation have been studied. Different plants: perennial ryegrass (Lolium perenne) and alfalfa (Medicago sativa); crops- maize (Zea mays L.) and bean (Phaseolus vulgaris); shrubs – privet (Ligustrum sempervirens) and trifoliate orange (Poncirus trifoliate); trees - poplar (Populus deltoides) and white mulberry (Morus alba L.) were exposed to hydrocarbons of different concentrations (1, 10 and 100 mM). Destructive changes in bean and maize leaves cells ultrastructure under the influence of benzene vapour were revealed at the level of photosynthetic and energy generation subcellular organells. Different deviations at the level of subcellular organelles structure and distribution were observed in alfalfa and ryegrass root cells under the influence of benzene and octane, absorbed through roots. The level of destructive changes is concentration dependent. Benzene at low 1 and 10 mM concentration caused the increase in glutamate dehydrogenase (GDH) activity in maize roots and leaves and in poplar and mulberry shoots, though to higher extent in case of lower, 1mM concentration. The induction was more intensive in plant roots. The highest tested 100mM concentration of benzene was inhibitory to the enzyme in all plants. Octane caused induction of GDH in all grassy plants at all tested concentrations; however the rate of induction decreased parallel to increase of the hydrocarbon concentration. Octane at concentration 1 mM caused induction of GDH in privet, trifoliate and white mulberry shoots. The highest, 100mM octane was characterized by inhibitory effect to GDH activity in all plants. Octane had inductive effect on malate dehydrogenase in almost all plants and tested concentrations, indicating the intensification of Trycarboxylic Acid Cycle. The data could be suggested for elaboration of criteria for plant selection for phytoremediation of oil hydrocarbons contaminated soils.

Keywords: Higher plants, hydrocarbons, cell ultrastructure, glutamate and malate dehydrogenases.

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60 Double Immobilized Lipase for the Kinetic Resolution of Secondary Alcohols

Authors: A. Ursoiu, C. Paul, C. Marcu, M. Ungurean, F. Péter

Abstract:

Sol-gel immobilization of enzymes, which can improve considerably their properties, is now one of the most used techniques. By deposition of the entrapped lipase on a solid support, a new and improved biocatalyst was obtained, which can be used with excellent results in acylation reactions. In this paper, lipase B from Candida antarctica was double immobilized on different adsorbents. These biocatalysts were employed in the kinetic resolution of several aliphatic secondary alcohols in organic medium. High total recovery yields of enzymatic activity, up to 560%, were obtained. For all the studied alcohols the enantiomeric ratios E were over 200. The influence of the reaction medium was studied for the kinetic resolution of 2-pentanol.

Keywords: Double immobilization, enantioselectivity, kineticresolution, lipase, racemates, sol-gel entrapment.

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59 The Effect of a Muscarinic Antagonist on the Lipase Activity

Authors: Zohreh Bayat, Dariush Minai-Tehrani

Abstract:

Lipases constitute one of the most important groups of industrial enzymes that catalyze the hydrolysis of triacylglycerol to glycerol and fatty acids. Muscarinic antagonist relieves smooth muscle spasm of the gastrointestinal tract and effect on the cardiovascular system. In this research the effect of a muscarinic antagonist on the lipase activity of Pseudomonas aeruginosa was studied. Lineweaver–Burk plot showed that the drug inhibited the enzyme by competitive inhibition. The IC50 value (0.16 mM) and Ki (0.03 mM) of the drug revealed the drug bound to enzyme with high affinity. Determination of enzyme activity in various pH and temperature showed that the maximum activity of lipase was at pH 8 and 60oC both in presence and absence of the drug.

Keywords: Bacteria, inhibition, kinetics, lipase.

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58 Determination of Alkaline Protease Production In Serratia Marcescens Sp7 Using Agro Wastes As Substrate Medium, Optimization Of Production Parameters And Purification Of The Enzyme

Authors: Baby Joseph, Sankarganesh Palaniyandi

Abstract:

The enzyme alkaline protease production was determined under solid state fermentation using the soil bacteria Serratia marcescens sp7. The maximum production was obtained from wheat bran medium than ground nut shell and chemically defined medium. The physiological fermentation factors such as pH of the medium (pH 8), Temperature (40oC) and incubation time (48 hrs) played a vital role in alkaline protease production in all the above. 100Mm NaCl has given better resolution during elution of the enzymes. The enzyme production was found to be associated with growth of the bacterial culture.

Keywords: Alkaline protease, Wheat bran, Ground nut shell, Serratia marcescens

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57 Bioactivity Evaluation of Cucurbitin Derived Enzymatic Hydrolysates

Authors: Ž. Vaštag, Lj. Popović, S. Popović

Abstract:

After cold pressing of pumpkin oil, the defatted oil cake (PUOC) was utilised as raw material for processing of bio-functional hydrolysates. In this study, the in vitro bioactivity of an alcalase (AH) and a pepsin hydrolysate (PH) prepared from the major pumpkin 12S globulin (cucurbitin) are compared. The hydrolysates were produced at optimum reaction conditions (temperature, pH) for the enzymes, during 60min. The bioactivity testing included antioxidant and angiotensin I converting enzyme inhibitory activity assays. The hydrolysates showed high potential as natural antioxidants and possibly antihypertensive agents in functional food or nutraceuticals. Additionally, preliminary studies have shown that both hydrolysates could exhibit modest α-amylase inhibitory activity, which indicates on their hypoglycemic potential.

Keywords: Cucurbitin, alcalase, pepsin, protein hydrolysates, in vitro bioactivity.

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56 Changes to Oxidative Stress Levels Following Exposure to Formaldehyde in Lymphocytes

Authors: Malinee Pongsavee

Abstract:

Formaldehyde is the illegal chemical substance used for food preservation in fish and vegetable. It can promote carcinogenesis. Superoxide dismutases are the important antioxidative enzymes that catalyze the dismutation of superoxide anion into oxygen and hydrogen peroxide. The resultant level of oxidative stress in formaldehyde-treated lymphocytes was investigated. The formaldehyde concentrations of 0, 20, 40, 60, 80 and 120μmol/L were treated in human lymphocytes for 12 hours. After 12 treated hours, the superoxide dismutase activity change was measured in formaldehyde-treated lymphocytes. The results showed that the formaldehyde concentrations of 60, 80 and 120μmol/L significantly decreased superoxide dismutase activities in lymphocytes (P < 0.05). The change of superoxide dismutase activity in formaldehyde-treated lymphocytes may be the biomarker for detect cellular injury, such as damage to DNA, due to formaldehyde exposure.

Keywords: Formaldehyde, lymphocytes, superoxide dismutase activity.

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55 Extracellular Laccase Production by Co-culture between Galactomyces reesii IFO 10823 and Filamentous Fungal Strains Isolated from Fungus Comb Using Natural Inducer

Authors: P. Chaijak, M. Lertworapreecha, C. Sukkasem

Abstract:

Extracellular laccases are copper-containing microbial enzymes with many industrial biotechnological applications. This study evaluated the ability of nutrients in coconut coir to enhance the yield of extracellular laccase of Galactomyces reesii IFO 10823 and develop a co-culture between this yeast and other filamentous fungi isolated from the fungus comb of Macrotermes sp. The co-culture between G. reesii IFO 10823 and M. indicus FJ-M-5 (G3) gave the highest activity at 580.20 U/mL. When grown in fermentation media prepared from coconut coir and distilled water at 70% of initial moisture without supplement addition, G3 produced extracellular laccase of 113.99 U/mL.

Keywords: Extracellular laccase, production, yeast, natural inducer.

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54 All Proteins Have a Basic Molecular Formula

Authors: Homa Torabizadeh

Abstract:

This study proposes a basic molecular formula for all proteins. A total of 10,739 proteins belonging to 9 different protein groups classified on the basis of their functions were selected randomly. They included enzymes, storage proteins, hormones, signalling proteins, structural proteins, transport proteins, immunoglobulins or antibodies, motor proteins and receptor proteins. After obtaining the protein molecular formula using the ProtParam tool, the H/C, N/C, O/C, and S/C ratios were determined for each randomly selected sample. In this case, H, N, O, and S coefficients were specified per carbon atom. Surprisingly, the results demonstrated that H, N, O, and S coefficients for all 10,739 proteins are similar and highly correlated. This study demonstrates that despite differences in the structure and function, all known proteins have a similar basic molecular formula CnH1.58 ± 0.015nN0.28 ± 0.005nO0.30 ± 0.007nS0.01 ± 0.002n. The total correlation between all coefficients was found to be 0.9999.

Keywords: Protein molecular formula, Basic unit formula, Protparam tool.

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53 The Oxidative Stress and the Antioxidant Defense of the Lower Vegetables towards an Environmental Pollution

Authors: Fadila Khaldi, Nedjoud Grara, Houria Berrebbah, Mohamed Réda Djebar

Abstract:

The use of bioindicators plants (lichens, bryophytes and Sphagnum....) in monitoring pollution by heavy metals has been the subject of several works. However, few studies have addressed the impact of specific type-s pollutants (fertilizers, pesticides.) on these organisms. We propose in this work to make the highlighting effect of NPKs (NPK: nitrogen-phosphate-potassium-sulfate (NP2O5K2O) (15,15,15), at concentrations of 10, 20, 30 , 40 and 50mM/L) on the activity of detoxification enzymes (GSH/GST, CAT, APX and MDA) of plant bioindicators (mosses and lichens) after treatment for 3 and 7 days. This study shows the important role of the defense system in the accumulation and tolerance to chemical pollutants through the activation of enzymatic (GST (glutathione-S-transferase, APX (ascorbat peroxidase), CAT (catalase)) and nonenzymatic biomarkers (GSH (glutathione), MDA (malondialdehyde)) against oxidative stress generated by the NPKs.

Keywords: NPKs, Bioindicators, lower plants, GSH / GST, CAT, APX and MDA.

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52 Biosynthesis of Silver-Phosphate Nanoparticles Using the Extracellular Polymeric Substance of Sporosarcina pasteurii

Authors: Mohammadhosein Rahimi, Mohammad Raouf Hosseini, Mehran Bakhshi, Alireza Baghbanan

Abstract:

Silver ions (Ag+) and their compounds are consequentially toxic to microorganisms, showing biocidal effects on many species of bacteria. Silver-phosphate (or silver orthophosphate) is one of these compounds, which is famous for its antimicrobial effect and catalysis application. In the present study, a green method was presented to synthesis silver-phosphate nanoparticles using Sporosarcina pasteurii. The composition of the biosynthesized nanoparticles was identified as Ag3PO4 using X-ray Diffraction (XRD) and Energy Dispersive Spectroscopy (EDS). Also, Fourier Transform Infrared (FTIR) spectroscopy showed that Ag3PO4 nanoparticles was synthesized in the presence of biosurfactants, enzymes, and proteins. In addition, UV-Vis adsorption of the produced colloidal suspension approved the results of XRD and FTIR analyses. Finally, Transmission Electron Microscope (TEM) images indicated that the size of the nanoparticles was about 20 nm.

Keywords: Bacteria, biosynthesis, silver-phosphate, Sporosarcina pasteurii, nanoparticle.

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51 An Algorithm of Regulation of Glucose-Insulin Concentration in the Blood

Authors: B. Selma, S. Chouraqui

Abstract:

The pancreas is an elongated organ that extends across the abdomen, below the stomach. In addition, it secretes certain enzymes that aid in food digestion. The pancreas also manufactures hormones responsible for regulating blood glucose levels. In the present paper, we propose a mathematical model to study the homeostasis of glucose and insulin in healthy human, and a simulation of this model, which depicts the physiological events after a meal, will be represented in ordinary humans. The aim of this paper is to design an algorithm which regulates the level of glucose in the blood. The algorithm applied the concept of expert system for performing an algorithm control in the form of an "active" used to prescribe the rate of insulin infusion. By decomposing the system into subsystems, we have developed parametric models of each subsystem by using a forcing function strategy. The results showed a performance of the control system.

Keywords: Modeling, algorithm, regulation, glucose-insulin, blood, control system.

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50 Plants and Microorganisms for Phytoremediation of Soils Polluted with Organochlorine Pesticides

Authors: Maritsa Kurashvili, George Adamia, Tamar Ananiashvili, Lia Amiranasvili, Tamar Varazi, Marina Pruidze, Marlen Gordeziani, Gia Khatisashvili

Abstract:

The goal of presented work is the development phytoremediation method targeted to cleaning environment polluted with organochlorine pesticides, based on joint application of plants and microorganisms. For this aim the selection of plants and microorganisms with corresponding capabilities towards three organochlorine pesticides (Lindane, DDT and PCP) has been carried out.

The tolerance of plants to tested pesticides and induction degree of plant detoxification enzymes by these compounds have been used as main criteria for estimating the applicability of plants in proposed technology. Obtained results show that alfalfa, maize and soybean among tested six plant species have highest tolerance to pesticides.

As a result of screening, more than 30 strains from genera Pseudomonas have been selected. As a result of GC analysis of incubation area, 11 active cultures for investigated pesticides are carefully chosen.

Keywords: DDT, Lindane, organochlorine pesticides, PCP, phytoremediation.

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49 Comparing Spontaneous Hydrolysis Rates of Activated Models of DNA and RNA

Authors: Mohamed S. Sasi, Adel M. Mlitan, Abdulfattah M. Alkherraz

Abstract:

This research project aims to investigate difference in relative rates concerning phosphoryl transfer relevant to biological catalysis of DNA and RNA in the pH-independent reactions. Activated Models of DNA and RNA for alkyl-aryl phosphate diesters (with 4-nitrophenyl as a good leaving group) have successfully been prepared to gather kinetic parameters. Eyring plots for the pH– independent hydrolysis of 1 and 2 were established at different temperatures in the range 100–160 °C. These measurements have been used to provide a better estimate for the difference in relative rates between the reactivity of DNA and RNA cleavage. Eyring plot gave an extrapolated rate of kH2O = 1 × 10-10 s -1 for 1 (RNA model) and 2 (DNA model) at 25°C. Comparing the reactivity of RNA model and DNA model shows that the difference in relative rates in the pH-independent reactions is surprisingly very similar at 25°. This allows us to obtain chemical insights into how biological catalysts such as enzymes may have evolved to perform their current functions.

Keywords: DNA & RNA Models, Relative Rates, Reactivity.

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48 Oxidantantioxidant Status in Calves Supplemented with Green Tea Extract

Authors: Ibrahim I. Elshahawy

Abstract:

The objective of the present study was to investigate the effect of green tea extract on serum oxidant and antioxidant profile, liver and kidney function. 40 Friesian calves are included in this study and allocated into two groups: Group I (n=20) clinically healthy calves showing no clinical abnormalities, not receiving any treatment and served as control; group II (n=20) received green tea extract (GTE) for 30 days. Non-significant changes in blood urea nitrogen (BUN) were detected between groups, on contrary, serum creatinine and activities of liver enzymes aspartate transaminase (AST) and alanine transaminase (ALT) were significantly different between two groups. There were significant increases in the mean values of serum antioxidative parameters (total antioxidant capacity, catalase, superoxide dismutase, reduced glutathione and glutathione peroxidase) in group II. Whereas, the activity of lipid peroxidase significantly decreased in GTE treated calves when compared to control.

Keywords: Green tea extract, antioxidants, oxidants, calves.

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47 Bioactive Component in Milk and Dairy Product

Authors: Bahareh Hajirostamloo

Abstract:

Recent research has shown that milk proteins can yield bioactive peptides with opioid, mineral binding, cytomodulatory, antihypertensive, immunostimulating, antimicrobial and antioxidative activity in the human body. Bioactive peptides are encrypted in milk proteins and are only released by enzymatic hydrolysis in vivo during gastrointestinal digestion, food processing or by microbial enzymes in fermented products. At present significant research is being undertaken on the health effects of bioactive peptides. A variety of naturally formed bioactive peptides have been found in fermented dairy products, such as yoghurt, sour milk and cheese. In particular, antihypertensive peptides have been identified in fermented milks, whey and ripened cheese. Some of these peptides have been commercialized in the form of fermented milks. Bioactive peptides have the potential to be used in the formulation of health-enhancing nutraceuticals, and as potent drugs with well defined pharmacological effects.

Keywords: Milk protein, Bioactive peptides, Health effects, Dairy product.

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46 Isolation and Characterization of Collagen from Chicken Feet

Authors: P. Hashim, M. S. Mohd Ridzwan, J. Bakar

Abstract:

Collagen was isolated from chicken feet by using papain and pepsin enzymes in acetic acid solution at 4°C for 24h with a yield of 18.16% and 22.94% by dry weight, respectively. Chemical composition and characteristics of chicken feet collagen such as amino acid composition, SDS-PAGE patterns, FTIR spectra and thermal properties were evaluated. The chicken feet collagen is rich in the amino acids glycine, glutamic acid, proline and hydroxyproline. Electrophoresis pattern demonstrated two distinct α-chains (α1 and α2) and β chain, indicating that type I collagen is a major component of chicken feet collagen. The thermal stability of collagen isolated by papain and pepsin revealed stable denaturation temperatures of 48.40 and 53.35°C, respectively. The FTIR spectra of both collagens were similar with amide regions in A, B, I, II and III. The study demonstrated that chicken feet collagen using papain isolation method is possible as commercial alternative ingredient. 

Keywords: Chicken feet, collagen, papain, pepsin.

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45 A Study of the Cyclic Variations of the Enzyme and the Electrolyte Activity in Uterine and Oviducal Secretions during an Estrous Cycle of the Ewe

Authors: Yahia M., Laanani I., Benbia S., Hachemi M., Massinissa Y.

Abstract:

Uterine and oviducal fluids are necessary for capacitation of the spermatozoa and early embryonic development. The aim of the present study was to determine the effects of estrous cycle phases (follicular and luteal) on some biological parameters (enzymes, electrolytes and total proteins) in uterine and oviducal secretions of ewes. Oviducal and uterine fluids were collected, diluted and centrifuged. According to our results, concentrations of GPT, G6PDH, total proteins, K and Na were significantly (P <0.05) higher at the luteal phase, however, the levels of aldolase, Mg, Ca and P were significantly (P <0.05) higher at the follicular phase in uterine secretions. While, only oviducal K and Ca were significantly (P<0.05) higher at the follicular phase. Our study revealed the existence of significant cyclic variations for some uterine and oviducal parameters which indicates the effect of ovarian hormones on the components of genital secretions.

Keywords: Biochemical parameters, estrous cycle, ewe, genital secretion.

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44 Vetiver Oil Production from Root Culture of Vetiveria zizanioides

Authors: Rizkita R. Esyanti, Iriawati, Olga Mardisadora

Abstract:

Vetiver oil is secondary metabolite that accumulates in Vetiveria zizanioides roots.  The aim of this study was to obtain best type of root culture which produce high amount of vetiver oil, and was similar to metabolites produce from its mother plant.  Protein analysis was also conducted to detect protein, related to putative enzymes, which have a role in terpenoids synthesis in the root culture. The results showed that root culture derived from crown explant produced the best root growth.   The root culture produced primary and lateral roots, ca. 40 branches.  The vetiver oil produced from root culture was analyzed by using GC-MS., and the highest content of terpenoids from roots of crown explant attained 19.024%.  The result of SDS PAGE showed proteins which were ±61 kD and ± 68 kD, each might be related to putative monoterpene synthase and sesquiterpenes complex, respectively. 

Keywords: Protein, Root culture, Terpenoids, Vetiver oil.

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43 Microfluidic Paper-Based Electrochemical Biosensor

Authors: Ahmad Manbohi, Seyyed Hamid Ahmadi

Abstract:

A low-cost paper-based microfluidic device (PAD) for the multiplex electrochemical determination of glucose, uric acid, and dopamine in biological fluids was developed. Using wax printing, PAD containing a central zone, six channels, and six detection zones was fabricated, and the electrodes were printed on detection zones using pre-made electrodes template. For each analyte, two detection zones were used. The carbon working electrode was coated with chitosan-BSA (and enzymes for glucose and uric acid). To detect glucose and uric acid, enzymatic reactions were employed. These reactions involve enzyme-catalyzed redox reactions of the analytes and produce free electrons for electrochemical measurement. Calibration curves were linear (R² > 0.980) in the range of 0-80 mM for glucose, 0.09–0.9 mM for dopamine, and 0–50 mM for uric acid, respectively. Blood samples were successfully analyzed by the proposed method.

Keywords: Multiplex, microfluidic paper-based electrochemical biosensors, biomarkers, biological fluids.

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42 The Effect of Selective Cyclooxygenase (COX) Inhibitors on Japanese Medaka (Oryzias latipes) Reproduction Parameters

Authors: Agata Kowalska, Radosław K. Kowalski, Zdzisław Zakęś

Abstract:

Our results showed that treatment with both cyclooxygenase (COX1 or COX2) inhibitors impair reproduction parameters of the medaka. Resveratrol (COX1 inhibitor) caused an decrease in the number of spawning females at the first week of feeding fish with experimental diets. In the group treated with NS- 398 (COX2 inhibitor) we found the lowest sperm velocity parameters and decreased linearity of movement. The ovaries of the medaka fed feed supplemented with Resveratrol or NS-398 were confirmed to have a lower share of matured oocytes however during the experiment (four weeks) the number of eggs spawned by females was similar. Both inhibitors in fish diet (20 mg/kg body weight/day) caused a decrease in the embryo survival. Our results revealed that for the medaka female reproduction, activity of both COX enzymes might be necessary whereas males reproduction competence, as expressed by sperm motility parameters, might be related to COX2 activity.

Keywords: COX innibitors, medaka, reproduction parameters

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