Search results for: cellulase enzyme

Authors: K. Ratanapongleka, J. Phetsom

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Extraction of laccase produced by L. polychrous in an aqueous two-phase system, composed of polyethylene glycol and phosphate salt at pH 7.0 and 250C was investigated. The effect of PEG molecular weight, PEG concentration and phosphate concentration was determined. Laccase preferentially partitioned to the top phase. Good extraction of laccase to the top phase was observed with PEG 4000. The optimum system was found in the system containing 12% w/w PEG 4000 and 16% w/w phosphate salt with KE of 88.3, purification factor of 3.0-fold and 99.1% yield. Some properties of the enzyme such as thermal stability, effect of heavy metal ions and kinetic constants were also presented in this work. The thermal stability decreased sharply with high temperature above 60 0C. The enzyme was inhibited by Cd2+, Pb2+, Zn2+ and Cu2+. The Vmax and Km values of the enzyme were 74.70 μmol/min/ml and 9.066 mM respectively.

Keywords: Aqueous Two Phase System, Laccase, Lentinuspolychrous,

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Authors: G. Baskar, C. Muthukumaran, S. Renganathan

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Enzymatic hydrolysis of starch from natural sources finds potential application in commercial production of alcoholic beverage and bioethanol. In this study the effect of starch concentration, temperature, time and enzyme concentration were studied and optimized for hydrolysis of cassava (Manihot esculenta) starch powder (of mesh 80/120) into glucose syrup by immobilized (using Polyacrylamide gel) a-amylase using central composite design. The experimental result on enzymatic hydrolysis of cassava starch was subjected to multiple linear regression analysis using MINITAB 14 software. Positive linear effect of starch concentration, enzyme concentration and time was observed on hydrolysis of cassava starch by a-amylase. The statistical significance of the model was validated by F-test for analysis of variance (p < 0.01). The optimum value of starch concentration temperature, time and enzyme concentration were found to be 4.5% (w/v), 45oC, 150 min, and 1% (w/v) enzyme. The maximum glucose yield at optimum condition was 5.17 mg/mL.

Keywords: Enzymatic hydrolysis, Alcoholic beverage, Centralcomposite design, Polynomial model, glucose yield.

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Authors: E. Ceyhan, A. Kahraman, M. Önder, M.K. Ateş, S. Karadaş, R. Topak, M.A. Avcı

Abstract:

The experimental design was 4 x 5 factorial with three replications in fully controlled research greenhouse in Department of Soil Sciences and Plant Nutrition, Faculty of Agriculture, University of Selcuk in the year of 2009. Determination of tolerant chickpea genotypes to drought was made in the research. Additionally, sophisticated effects of drought on plant growth and development, biochemical and physical properties or physical defense mechanisms were presented. According to the results, the primary genotypes were Ilgın YP (0.0063 g/gh) for leaf water capacity, 22235 70.44(%) for relative water content, 22159 (82.47%) for real water content, 22159 (5.03 mg/l) for chlorophyll a+b, Ilgın YP (125.89 nmol H2O2.dak-1/ mg protein-1) for peroxidase, Yunak YP (769.67 unit/ mg protein-1) for superoxide dismutase, Seydişehir YP (16.74 μg.TA-1) for proline, Gökçe (80.01 nmol H2O2.dak-1/ mg protein-1) for catalase. Consequently, all the genotypes increased their enzyme activity depending on the increasing of drought stress consider with the effects of drought stress on leaf enzyme activity. Chickpea genotypes are increasing enzyme activity against to drought stress.

Keywords: Chickpea, drought, enzyme, tolerance to drought

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

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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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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: Homa Torabizadeh, Mohaddeseh Mikani

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Inulinase from Aspergillus niger was covalently immobilized on magnetic nanoparticles (MNPs/Fe₃O₄) covered with soy protein isolate (SPI/Fe₃O₄) functionalized by bovine serum albumin (BSA) nanoparticles. MNPs are promising enzyme carriers because they separate easily under external magnetic fields and have enhanced immobilized enzyme reusability. As MNPs aggregate simply, surface coating strategy was employed. SPI functionalized by BSA was a suitable candidate for nanomagnetite coating due to its superior biocompatibility and hydrophilicity. Fe₃O₄@SPI-BSA nanoparticles were synthesized as a novel carrier with narrow particle size distribution. Step by step fabrication monitoring of Fe₃O₄@SPI-BSA nanoparticles was performed using field emission scanning electron microscopy and dynamic light scattering. The results illustrated that nanomagnetite with the spherical morphology was well monodispersed with the diameter of about 35 nm. The average size of the SPI-BSA nanoparticles was 80 to 90 nm, and their zeta potential was around −34 mV. Finally, the mean diameter of fabricated Fe₃O₄@SPI-BSA NPs was less than 120 nm. Inulinase enzyme from Aspergillus niger was covalently immobilized through gluteraldehyde on Fe₃O₄@SPI-BSA nanoparticles successfully. Fourier transform infrared spectra and field emission scanning electron microscopy images provided sufficient proof for the enzyme immobilization on the nanoparticles with 80% enzyme loading.

Keywords: High fructose syrup, inulinase immobilization, functionalized magnetic nanoparticles, soy protein isolate.

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Authors: Khalida Boutemak, Nasssima Benali, Nadji Moulai-Mostefa

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Effect of enzyme on the yield and chemical composition of Artemisia campestris essential oil is reported in the present study. It was demonstrated that enzyme facilitated the extraction of essential oil with increase in oil yield and did not affect any noticeable change in flavour profile of the  volatile oil. Essential oil was tested for antibacterial activity using Escherichia coli; which was extremely sensitive against control with the largest inhibition (29mm), whereas Staphylococcus aureus was the most sensitive against essential oil obtained from enzymatic pre-treatment with the largest inhibition zone (25mm). The antioxidant activity of the essential oil with hemicellulase pre-treatment (EO2) and control sample (EO1) was determined through reducing power. It was significantly lower than the standard drug (vitamin C) in this order: vitamin C˃EO2˃EO1.

Keywords: Artemisia campestris, enzyme pre-treatment, hemicellulase, antibacterial activity, antioxidant activity.

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Authors: M. Yaldagard, S.A. Mortazavi, F. Tabatabaie

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In the present study, the effects of ultrasound as emerging technology were investigated on germination stimulation, amount of alpha-amylase activity on dry barley seeds before steeping stage of malting process. All experiments were carried out at 20 KHz on the ultrasonic generator in 3 different ultrasonic intensities (20, 60 and 100% setting from total power of device) and time (5, 10 and 15 min) at constant temperature (30C). For determining the effects of these parameters on enzyme the Fuwa method assay based on the decreased staining value of blue starch–iodine complexes employed for measurement an activity. The results of these assays were analyzed by Qualitek4 software using the Taguchi statistical method to evaluate the factor-s effects on enzyme activity. It has been found that when malting barley is irradiated with an ultrasonic power, a stimulating effect occurs as to the enzyme activity.

Keywords: ultrasound, alpha-amylase activity, stimulationand Taguchi statistical method.

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

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

Keywords: Cheese, Papain, polyurethane, Staphylococcus aureus.

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

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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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Authors: P. Pinyaphong, S. Phutrakul

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

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

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Authors: Anna Trusek-Holownia, Andrzej Noworyta

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Using an enzyme of known specificity the hydrolysis of protein was carried out in a controlled manner. The aim was to obtain oligopeptides being the so-called active peptides or their direct precursors. An original way of expression of the protein hydrolysis kinetics was introduced. Peptide bonds contained in the protein were recognized as a diverse-quality substrate for hydrolysis by the applied protease. This assumption was positively verified taking as an example the hydrolysis of albumin by thermolysin. Peptide linkages for this system should be divided into at least four groups. One of them is a group of bonds non-hydrolyzable by this enzyme. These that are broken are hydrolyzed at a rate that differs even by tens of thousands of times. Designated kinetic constants were k'_F = 10991.4 L/g^.h, k'_M = 14.83L/g^.h, k'_S about 10^-1 L/g^.h for fast, medium and slow bonds, respectively. Moreover, a procedure for unfolding of the protein, conducive to the improved susceptibility to enzymatic hydrolysis (approximately three-fold increase in the rate) was proposed.

Keywords: Peptide bond hydrolysis, kinetics, enzyme specificity, biologically active peptides.

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

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

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

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Authors: Ndidi F. Amulu, Calistus N. Ude, Patrick E. Amulu, Nneka N. Uchegbu

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The effects of temperature and enzyme concentration on the quality of mixed pineapple and pawpaw blended fruits juice were studied. Extracts of the two fruit juices were separately treated at 70  for 15 min each so as to inactivate micro-organisms. They were analyzed and blended in different proportions of 70% pawpaw and 30% pineapple, 60% pawpaw and 40% pineapple, 50% pineapple and 50% pawpaw, 40% pawpaw and 60% pineapple. The characterization of the fresh pawpaw and pineapple juice before blending showed that the juices have good quality. The high water content of the product may have affected the viscosity, vitamin C content and total soluble solid of the blended juice to be low. The effects of the process parameters on the quality showed that better quality of the blended juice can be obtained within the optimum temperature range of (50-70 °C) and enzyme concentration range (0.12-0.18 w/v). The ratio of mix 60% pineapple juice: 40% pawpaw juice has better quality. This showed that pawpaw and pineapple juices can blend effectively to produce a quality juice.

Keywords: Clarification, pawpaw, pineapple, viscosity, vitamin C.

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Authors: P. Pinyaphong, P. Sriburi, S. Phutrakul

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This paper studied the synthesis of monoacylglycerol (monolaurin) by glycerolysis of coconut oil and crude glycerol, catalyzed by Carica papaya lipase. Coconut oil obtained from cold pressed extraction method and crude glycerol obtained from the biodiesel plant in Department of Chemistry, Uttaradit Rajabhat University, Thailand which used oils were used as raw materials for biodiesel production through transesterification process catalyzed by sodium hydroxide. The influences of the following variables were studied: (i) type of organic solvent, (ii) molar ratio of substrate, (iii) reaction temperature, (iv) reaction time, (v) lipase dosage, and (vi) initial water activity of enzyme. High yields in monoacylglycerol (58.35%) were obtained with molar ratio of glycerol to oil at 8:1 in ethanol, temperature was controlled at 45oC for 36 hours, the amount of enzyme used was 20 wt% of oil and initial water activity of enzyme at 0.53.

Keywords: Monoacylglycerol, crude glycerol, coconut oil, Carica papaya lipase.

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Authors: Shweta Kumari, Parmjit S. Panesar, Manab B. Bera

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The hydrolysis of lactose using β-galactosidase is one of the most promising biotechnological applications, which has wide range of potential applications in food processing industries. However, due to intracellular location of the yeast enzyme, and expensive extraction methods, the industrial applications of enzymatic hydrolysis processes are being hampered. The use of permeabilization technique can help to overcome the problems associated with enzyme extraction and purification of yeast cells and to develop the economically viable process for the utilization of whole cell biocatalysts in food industries. In the present investigation, standardization of permeabilization process of novel yeast isolate was carried out using a statistical model approach known as Response Surface Methodology (RSM) to achieve maximal b-galactosidase activity. The optimum operating conditions for permeabilization process for optimal β-galactosidase activity obtained by RSM were 1:1 ratio of toluene (25%, v/v) and ethanol (50%, v/v), 25.0 ^oC temperature and treatment time of 12 min, which displayed enzyme activity of 1.71 IU /mg DW.

Keywords: β-galactosidase, optimization, permeabilization, response surface methodology, yeast.

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Authors: K. Manikandan, T. Viruthagiri

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Studies on Simultaneous Saccharification and Fermentation (SSF) of corn flour, a major agricultural product as the substrate using starch digesting glucoamylase enzyme derived from Aspergillus niger and non starch digesting and sugar fermenting Saccharomyces cerevisiae in a batch fermentation. Experiments based on Central Composite Design (CCD) were conducted to study the effect of substrate concentration, pH, temperature, enzyme concentration on Ethanol Concentration and the above parameters were optimized using Response Surface Methodology (RSM). The optimum values of substrate concentration, pH, temperature and enzyme concentration were found to be 160 g/l, 5.5, 30°C and 50 IU respectively. The effect of inoculums age on ethanol concentration was also investigated. The corn flour solution equivalent to 16% initial starch concentration gave the highest ethanol concentration of 63.04 g/l after 48 h of fermentation at optimum conditions of pH and temperature. Monod model and Logistic model were used for growth kinetics and Leudeking – Piret model was used for product formation kinetics.

Keywords: Simultaneous Saccharification and Fermentation(SSF), Corn Starch, Ethanol, Logisitic Model.

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Authors: Govindaraju.M, Ganeshkumar.R.S, Suganthi.P, Muthukumaran.V.R, Visvanathan.P

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The present study was conducted to investigate the response of plants exposed to lignite-based thermal power plant emission. For this purpose, five plant species were collected from 1.0 km distance (polluted site) and control plants were collected from 20.0 km distance (control site) to thermal power plant. The common tree species Cassia siamea Lamk., Polyalthia longifolia. Sonn, Acacia longifolia (Andrews) Wild., Azadirachta indica A.Juss, Ficus religiosa L. were selected as test plants. Photosynthetic pigments changes (chlorophyll a, chlorophyll b and carotenoids) and rubisco enzyme modifications were studied. Reduction was observed in the photosynthetic pigments of plants growing in polluted site and also large sub unit of the rubisco enzyme was degraded in Azadirachta indica A. Juss collected from polluted site.

Keywords: Air pollution, Lignite-based thermal power plant, Photosynthetic pigments, Rubisco enzyme.

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Authors: E. Danso-Boateng

Abstract:

The effects of enzyme action and heat pretreatment on oil extraction yield from sunflower kernels were analysed using hexane extraction with Soxhlet, and aqueous extraction with incubator shaker. Ground kernels of raw and heat treated kernels, each with and without Viscozyme treatment were used. Microscopic images of the kernels were taken to analyse the visible effects of each treatment on the cotyledon cell structure of the kernels. Heat pretreated kernels before both extraction processes produced enhanced oil extraction yields than the control, with steam explosion the most efficient. In hexane extraction, applying a combination of steam explosion and Viscozyme treatments to the kernels before the extraction gave the maximum oil extractable in 1 hour; while for aqueous extraction, raw kernels treated with Viscozyme gave the highest oil extraction yield. Remarkable cotyledon cell disruption was evident in kernels treated with Viscozyme; whereas steam explosion and conventional heat treated kernels had similar effects.

Keywords: Enzyme-assisted aqueous and hexane extraction, heatpretreatment, sunflower cotyledon structure, sunflower oil extraction

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Authors: Namsoo Kim, So-Hee Son, Jin-Soo Maeng, Yong-Jin Cho, Chul-Jin Kim, Chong-Tai Kim

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Wheat gluten hydrolyzates (WGHs) and anchovy fine powder hydrolyzates (AFPHs) were produced at 300 MPa using combinations of Flavourzyme 500MG (F), Alcalase 2.4L (A), Marugoto E (M) and Protamex (P), and then were compared to those produced at ambient pressure concerning the contents of soluble solid (SS), soluble nitrogen and electrophoretic profiles. The contents of SS in the WGHs and AFPHs increased up to 87.2% according to the increase in enzyme number both at high and ambient pressure. Based on SS content, the optimum enzyme combinations for one-, two-, three- and four-enzyme hydrolysis were determined as F, FA, FAM and FAMP, respectively. Similar trends were found for the contents of total soluble nitrogen (TSN) and TCA-soluble nitrogen (TCASN). The contents of SS, TSN and TCASN in the hydrolyzates together with electrophoretic mobility maps indicates that the high-pressure treatment of this study accelerated protein hydrolysis compared to ambient-pressure treatment.

Keywords: Production, Wheat gluten hydrolyzates, Anchovy fine powder hydrolyzates, Protease combinations.

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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: Pantip Kayee

Abstract:

17α-ethinylestradiol (EE₂) is a recalcitrant micropollutant which is found in small amounts in municipal wastewater. But these small amounts still adversely affect for the reproductive function of aquatic organisms. Evidence in the past suggested that full-scale WWTPs equipped with nitrification process enhanced the removal of EE₂ in the municipal wastewater. EE₂ has been proven to be able to be transformed by ammonia oxidizing bacteria (AOB) via co-metabolism. This research aims to clarify the EE₂ degradation pattern by different consortium of ammonia oxidizing microorganism (AOM) including AOA (ammonia oxidizing archaea) and investigate contribution between the existing ammonia monooxygenase (AMO) and new synthesized AOM. The result showed that AOA or AOB of N. oligotropha cluster in enriched nitrifying activated sludge (NAS) from 2mM and 5mM, commonly found in municipal WWTPs, could degrade EE₂ in wastewater via co-metabolism. Moreover, the investigation of the contribution between the existing ammonia monooxygenase (AMO) and new synthesized AOM demonstrated that the new synthesized AMO enzyme may perform ammonia oxidation rather than the existing AMO enzyme or the existing AMO enzyme may has a small amount to oxidize ammonia.

Keywords: 17α-ethinylestradiol, nitrification, ammonia oxidizing bacteria, ammonia oxidizing archaea.

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Authors: N.Peatciyammal, B.Balachandar, M.Dinesh Kumar, K.Tamilarasan, C.Muthukumaran

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Enzymatic hydrolysis of starch from natural sources finds potential application in commercial production of alcoholic beverage and bioethanol. In this study the effect of starch concentration, temperature, time and enzyme concentration were studied and optimized for hydrolysis of Potato starch powder (of mesh 80/120) into glucose syrup by immobilized (using Sodium arginate) α-amylase using central composite design. The experimental result on enzymatic hydrolysis of Potato starch was subjected to multiple linear regression analysis using MINITAB 14 software. Positive linear effect of starch concentration, enzyme concentration and time was observed on hydrolysis of Potato starch by α-amylase. The statistical significance of the model was validated by F-test for analysis of variance (p ≤ 0.01). The optimum value of starch concentration, enzyme concentration, temperature, time and were found to be 6% (w/v), 2% (w/v), 40°C and 80min respectively. The maximum glucose yield at optimum condition was 2.34 mg/mL.

Keywords: Alcoholic beverage, Central Composite Design, Enzymatic hydrolysis, Glucose yield, Potato Starch.

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Authors: G. Arabaci

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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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Authors: Seung Pil Pack

Abstract:

Carbonic anhydrases (CAs) has been focused as biological catalysis for CO2 sequestration process because it can catalyze the conversion of CO2 to bicarbonate. Here, codon-optimized sequence of α type-CA cloned from Duneliala species. (DsCAopt) was constructed, expressed, and characterized. The expression level in E. coli BL21(DE3) was better for codon-optimized DsCAopt than intact sequence of DsCAopt. DsCAopt enzyme shows high-stability at pH 7.6/10.0. In final, we demonstrated that in the Ca2+ solution, DsCAopt enzyme can catalyze well the conversion of CO2 to CaCO3, as the calcite form.

Keywords: Carbonic anhydrase, Codon-optimization, Duneliala species, CO2 sequestration

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Authors: F. Yilmaz, Y. Saylan, A. Derazshamshir, S. Atay, A. Denizli

Abstract:

A quartz crystal microbalance (QCM) nanosensor was developed to detect lysozyme enzyme by functionalizing its gold surface with the attachment of poly(methacroyl-L-phenylalanine) (PMAPA) nanoparticles. PMAPA was chosen as a hydrophobic matrix. The hydrophobic nanoparticles were synthesized by micro-emulsion polymerization method. Hydrophobic QCM nanosensor was tested for real time detection of lysozyme enzyme from aqueous solution. The kinetic and affinity studies were determined by using lysozyme solutions with different concentrations. The responses related with mass (Δm) and frequency (Δf) shifts were used to evaluate adsorption properties. 

 

Keywords: HIC, lysozyme, nanosensor, QCM.

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Authors: S. S. Rana, C. Janveja, S. K. Soni

Abstract:

This study is concerned with the optimization of fermentation parameters for the hyper production of mannanase from Fusarium oxysporum SS-25 employing two step statistical strategy and kinetic characterization of crude enzyme preparation. The Plackett-Burman design used to screen out the important factors in the culture medium revealed 20% (w/w) wheat bran, 2% (w/w) each of potato peels, soyabean meal and malt extract, 1% tryptone, 0.14% NH4SO4, 0.2% KH2PO4, 0.0002% ZnSO4, 0.0005% FeSO4, 0.01% MnSO4, 0.012% SDS, 0.03% NH4Cl, 0.1% NaNO3 in brewer’s spent grain based medium with 50% moisture content, inoculated with 2.8×107 spores and incubated at 30oC for 6 days to be the main parameters influencing the enzyme production. Of these factors, four variables including soyabean meal, FeSO4, MnSO4 and NaNO3 were chosen to study the interactive effects and their optimum levels in central composite design of response surface methodology with the final mannanase yield of 193 IU/gds. The kinetic characterization revealed the crude enzyme to be active over broader temperature and pH range. This could result in 26.6% reduction in kappa number with 4.93% higher tear index and 1% increase in brightness when used to treat the wheat straw based kraft pulp. The hydrolytic potential of enzyme was also demonstrated on both locust bean gum and guar gum.

Keywords: Brewer’s Spent Grain, Fusarium oxysporum, Mannanase, Response Surface Methodology.

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Authors: Dyah Iswantini, Trivadila, Novik Nurhidayat, Waras Nurcholis

Abstract:

The antioxidant compounds are needed for the food, beverages, and pharmaceuticals industry. For this purpose, an appropriate method is required to measure the antioxidant properties in various types of samples. Spectrophotometric method usually used has some weaknesses, including the high price, long sample preparation time, and less sensitivity. Among the alternative methods developed to overcome these weaknesses is antioxidant biosensor based on superoxide dismutase (SOD) enzyme. Therefore, this study was carried out to measure the SOD activity originating from Deinococcus radiodurans and to determine its kinetics properties. Carbon paste electrode modified with ferrocene and immobilized SOD exhibited anode and cathode current peak at potential of +400 and +300mv respectively, in both pure SOD and SOD of D. radiodurans. This indicated that the current generated was from superoxide catalytic dismutation reaction by SOD. Optimum conditions for SOD activity was at pH 9 and temperature of 27.50C for D. radiodurans SOD, and pH 11 and temperature of 200C for pure SOD. Dismutation reaction kinetics of superoxide catalyzed by SOD followed the Lineweaver-Burk kinetics with D. radiodurans SOD KMapp value was smaller than pure SOD. The result showed that D. radiodurans SOD had higher enzyme-substrate affinity and specificity than pure SOD. It concluded that D. radiodurans SOD had a great potential as biological recognition component for antioxidant biosensor.

Keywords: Antioxidant biosensor, Deinococcus radiodurans, enzyme kinetic, superoxide dismutase (SOD).

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Authors: Fatima Zohra Ibn Majdoub Hassani, Ivan Lavandera, Joseph Kreit

Abstract:

This study explored the effects of different organic solvents, temperature, and the amount of glycerol on the alcohol dehydrogenase (ADH)-catalysed stereoselective reduction of different ketones. These conversions were then analyzed by gas chromatography. It was found that when the amount of deep eutectic solvents (DES) increases, it can improve the stereoselectivity of the enzyme although reducing its ability to convert the substrate into the corresponding alcohol. Moreover, glycerol was found to have a strong stabilizing effect on the ADH from Ralstonia sp. (E. coli/ RasADH). In the case of organic solvents, it was observed that the best conversions into the alcohols were achieved with DMSO and hexane. It was also observed that temperature decreased the ability of the enzyme to convert the substrates into the products and also affected the selectivity. In addition to that, the recycling of DES up to three times gave good conversions and enantiomeric excess results and glycerol showed a positive effect in the stability of various ADHs. Using RasADH, a good conversion and enantiomeric excess into the S-alcohol were obtained. It was found that an enhancement of the temperature disabled the stabilizing effect of glycerol and decreased the stereoselectivity of the enzyme. However, for other ADHs a temperature increase had an opposite positive effect, especially with ADH-T from Thermoanaerobium sp. One of the objectives of this study was to see the effect of cofactors such as NAD(P) on the biocatlysis activities of ADHs.

Keywords: Alcohol dehydrogenases, DES, gas chromatography, RasADH.

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