Search results for: enzyme activities

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: 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: J. Kumchai, J. Z. Huang, C. Y. Lee, F. C. Chen, S. W. Chin

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Cabbage seedlings grown in vitro were exposed to excess levels of heavy metals, including Cd, Mo, and Zn. High metal levels affected plant growth at cotyledonary stage. Seedlings under Cd, Mo, and Zn treatments could not produce root hairs and true leaves. Under stress conditions, seedlings accumulated a higher amount of anthocyanins in their cotyledons than those in the control. The pigments isolated from Cd and Zn stressed seedling cotyledons appeared as pink, while under Mo stress, was dark pink or purple. Moreover, excess Mo stress increased antioxidant enzyme activities of APX, CAT, SOD. These results suggest that, under excess Mo stress, the induced antioxidant enzyme activity of cabbage seedlings may function as a protective mechanism to shield the plants from toxicity and exacerbated growth.

Keywords: Anthocyanin, antioxidant enzyme activity, heavy metal, growth inhibition.

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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: Dayana Amira R., Roshanida A.R., Rosli M.I.

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Empty Fruit Bunches (EFB) and Palm Oil Mill Effluent (POME) are two main wastes from oil palm industries which contain rich lignocellulose. Degradation of EFB and POME by microorganisms will produce hydrolytic enzyme which will degrade cellulose and hemicellulose during composting process. However, normal composting takes about four to six months to reach maturity. Hence, application of fungi into compost can shorten the period of composting. This study identifies the effect of xylanase and cellulase produced by Aspergillus niger and Trichoderma virens on composting process using EFB and POME. The degradation of EFB and POME indicates the lignocellulolytic capacity of Aspergillus niger and Trichoderma virens with more than 7% decrease in hemicellulose and more than 25% decrease in cellulose for both inoculated compost. Inoculation of Aspergillus niger and Trichoderma virens also increased the enzyme activities during the composting period compared to the control compost by 21% for both xylanase and cellulase. Rapid rise in the activities of cellulase and xylanase was observed by Aspergillus niger with the highest activities of 14.41 FPU/mg and 3.89 IU/mg, respectively. Increased activities of cellulase and xylanase also occurred in inoculation of Trichoderma virens with the highest activities obtained at 13.21 FPU/mg and 4.43 IU/mg, respectively. Therefore, it is evident that the inoculation of fungi can increase the enzyme activities hence effectively degrading the EFB and POME.

Keywords: EFB, cellulase, POME, xylanase

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Authors: Fu E. Tang, Chung W. Tong

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“Garbage enzyme", a fermentation product of kitchen waste, water and brown sugar, is claimed in the media as a multipurpose solution for household and agricultural uses. This study assesses the effects of dilutions (5% to 75%) of garbage enzyme in reducing pollutants in domestic wastewater. The pH of the garbage enzyme was found to be 3.5, BOD concentration about 150 mg/L. Test results showed that the garbage enzyme raised the wastewater-s BOD in proportion to its dilution due to its high organic content. For mixtures with more than 10% garbage enzyme, its pH remained acidic after the 5-day digestion period. However, it seems that ammonia nitrogen and phosphorus could be removed by the addition of the garbage enzyme. The most economic solution for removal of ammonia nitrogen and phosphorus was found to be 9%. Further tests are required to understand the removal mechanisms of the ammonia nitrogen and phosphorus.

Keywords: Wastewater treatment, garbage enzyme, wastewater additives, ammonia nitrogen, phosphorus.

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Authors: K. Vasupen, S. Wongsuthavas, S. Bureenok, B. Saenmahayak, K. Ampaporn, C. Yuangklang

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This study was designed to determine effect of supplemented tomato pomace and fobrolytic enzyme on egg production and egg quality. A total of 40 CP brown laying hens (95 week old) were used in completely randomized design in 2x2 factorial arrangement with or without enzyme supplementation. Four dietary treatments included: Control (C), Fibrolytic enzyme (FE), 10% Tomato pomace (TP), and Fibrolytic enzyme + 10 % Tomato pomace (FE+TP). Each of the four dietary treatments was fed up to 30 days (10 birds/treatment). Live performance, egg production, egg weight and quality were determined for whole period. Dietary treatments had no effect (P>0.05) on live performance, egg weight, yolk color, and egg production. Therefore, laying hens fed diets with fibrolytic enzyme were significantly (P<0.05) increased yolk weight (17.37 g) as compared to other treatments. Additional of dietary tomato pomace had reduced capital costs for egg production.

Keywords: Hen, Tomato Pomace, Fibrolytic Enzyme, Egg Quality.

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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: 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: Mustafa Erat

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Glutathione S-transferase was purified from human erythrocytes and effects of some polyphenols were investigated on the enzyme activity. The purification procedure was performed on Glutathione-Agarose affinity chromatography after preparation of erythrocytes hemolysate with a yield of 81%. The purified enzyme showed a single band on the SDS-PAGE. The effects of some poliphenolic compounds such as catechin, dopa, dopamine, progallol and catechol were examined on the in vitro GST activity. Catechin was determined to be inhibitor for the enzyme, but others were not effective on the enzyme as inhibitors or activators. IC50 value -the concentration of inhibitor which reduces enzyme activity by 50%- was estimated to be 10 mM. Ki constants were also calculated as 6.38 ± 0,70 mM with GSH substrate, and 3.86 ± 0,78 mM with CDNB substrate using the equations of graphs for the inhibitor, and its inhibition type was determined as non-competitive.

Keywords: Drug resistance, Glutathione S-transferase, Inhibition.

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Authors: Zhanar S. Kudiyarova, Zhanar K. Rakhmetova, L. K. Bekbayeva, N. Z. Omirbekova, M. K. Gilmanov

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Malate dehydrogenase-glutamate oxaloacetate aminotransferase (MDh-GOAT) enzyme complex (the EC) was isolated and purified from wheat and rise, their some main physicchemical properties were studied. Michael-s constants of the EC MDh-GOAT to malate, glutamate and NAD were investigated. This kinetic results show a high relationship to glutamate. Taking into account important role of the the EC in catabolism of glutamate – the central amino acid of a nitric exchange, there is a sharp necessity of deeper studying of this enzyme complex. Therefore the basic purpose of the work is studying the basic physical and chemical properties of this enzyme complex discovered by us, which would be very important for understanding the mechanisms of reaction catalyzed by the EC.

Keywords: Malate dehydrogenase-glutamate oxaloacetate aminotransferase, enzyme complex, glutamate.

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Authors: R. Majidzadeh Heravi, M. Sankian, H. Kermanshahi, M. R. Nassiri, A. Heravi Moussavi, S. A. Lari, A. R. Varasteh

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The use of probiotics engineered to express specific enzymes has been the subject of considerable attention in poultry industry because of increased nutrient availability and reduced cost of enzyme supplementation. Phytase enzyme is commonly added to poultry feed to improve digestibility and availability of phosphorus from plant sources. To construct a probiotic with potential of phytate degradation, phytase gene (appA) from E. coli was cloned and transformed into two probiotic bacteria Lactobacillus salivarius and Lactococcus lactis. L. salivarous showed plasmid instability, unable to express the gene. The expression of appA gene in L. lactis was analyzed by detecting specific RNA and zymography assay. Phytase enzyme was isolated from cellular extracts of recombinant L. lactis, showing a 46 kDa band upon the SDS-PAGE analysis. Zymogram also confirmed the phytase activity of the 46 kDa band corresponding to the enzyme. An enzyme activity of 4.9U/ml was obtained in cell extracts of L. lactis. The growth of native and recombinant L. lactis was similar in the presence of two concentrations of ox bile.

Keywords: Lactobacillus salivarus, Lactococcus lactis, recombinant, phytase, poultry.

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

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

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

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

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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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Authors: Fazlollah Moosavinasab, Zhila Motamedi

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

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

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

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

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

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Authors: Zohreh Bayat, Dariush Minai-Tehrani

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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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Authors: W. Wanmolee, W. Sornlake, N. Laosiripojana, V. Champreda

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Conversion of lignocellulosic biomass is the basis process for production of fuels, chemicals and materials in the sustainable biorefinery industry. Saccharification of lignocellulosic biomass is an essential step which produces sugars for further conversion to target value-added products e.g. bio-ethanol, bio-plastic, g-valerolactone (GVL), 5-hydroxymethylfuroic acid (HMF), levulinic acid, etc. The goal of this work was to develop an efficient enzyme for conversion of biomass to reducing sugar based on crude fungal enzyme from Chaetomium globosum BCC5776 produced by submerged fermentation and evaluate its activity comparing to a commercial Acremonium cellulase. Five local biomasses in Thailand: rice straw, sugarcane bagasse, corncobs, corn stovers, and palm empty fruit bunches were pretreated and hydrolyzed with varying enzyme loadings. Saccharification of the biomass led to different reducing sugar levels from 115 mg/g to 720 mg/g from different types of biomass using cellulase dosage of 9 FPU/g. The reducing sugar will be further employed as sugar feedstock for production of ethanol or commodity chemicals. This work demonstrated the use of promising enzyme candidate for conversion of local lignocellulosic biomass in biorefinery industry.

Keywords: Biomass, Cellulase, Chaetomiun glubosum, Saccharification.

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

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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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Authors: Baby Joseph, Sankarganesh Palaniyandi

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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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Authors: F. Bamdad, Sh. Dokhani, J. Keramat, R. Zareie

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Biologically active peptides are of particular interest in food science and human nutrition because they have been shown to play several physiological roles. In vitro gastrointestinal digestion of lentil and whey proteins in this study produced high angiotensin-I converting enzyme inhibitory activity with 75.5±1.9 and 91.4±2.3% inhibition, respectively. High ACE inhibitory activity was observed in lentil after 5 days of germination (84.3±1.2%). Fractionation by reverse phase chromatography gave inhibitory activities as high as 86.3±2.0 for lentil, 94.8±1.8% for whey and 93.7±1.7% at 5th day of germination. Further purification by HPLC resulted in several inhibitory peptides with IC50 values ranging from 0.064 to 0.164 mg/ml. These results demonstrate that lentil proteins are a good source of peptides with ACE inhibitory activity that can be released by germination or gastrointestinal digestion. Despite the lower bioactivity in comparison with whey proteins, incorporation of lentil proteins in functional food formulations and natural drugs look promising.

Keywords: ACE inhibitory peptides, digestion, germination, lentil proteins, whey proteins

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Authors: N. Hachemi, A. Nouani, A. Benchabane

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The influence of cultivation factors such as content of ammonium sulfate, glucose and water in the culture medium and particle size of dry orange waste, on their bioconversion for pectinase production was studied using complete factorial design. A polygalacturonase (PG) was isolated using ion exchange chromatography under gradient elution 0-0,5 m/l NaCl (column equilibrate with acetate buffer pH 4,5), subsequently by sephadex G75 column chromatography was applied and the molecular weight was obtained about 51,28 KDa. Purified PG enzyme exhibits a pH and temperature optima of activity at 5 and 35°C respectively. Treatment of apple juice by purified enzyme extract yielded a clear juice, which was competitive with juice yielded by pure Sigma Aldrich Aspergillus niger enzyme.

Keywords: Bioconversion, orange wastes, optimization, pectinase.

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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

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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Authors: J. Salaenoi, A. Thongpan, M. Mingmuang

Abstract:

The activities of alkaline phosphatase and Ca2+ATPase in mud crab (Scylla serrata) collected from a soft-shell crab farm in Chantaburi Province, Thailand, in several stages of molting cycle were observed. The results showed that the activity of alkaline phosphatase in gill after molting was highly significant (p<0.05) comparing to those at intermolt and premolt stages. The activity profiles of alkaline phosphatase in integument and haemolymph were similar showing a decrease from intermolt to 2- week premolt stage and increased during 2-day premolt to 6-h postmolt stage before dropping at 7-day postmolt stage, while this enzyme in the gill was quite low at intermolt and premolt stages. For Ca2+ATPase, the activity profiles in gill and integument corresponded to the molting variation, especially the activities increased during 5-7 day postmolt stage were at highly significant levels (p<0.05) comparing to those at premolt and early postmolt stages. The highest activity of Ca2+ATPase in haemolymph was found at 2-week premolt stage (p<0.05). Changes in alkaline phosphatase and Ca2+ATPase activities over the molting cycle clearly indicated their active functions on calcification.

Keywords: Scylla serrata, molting cycle, alkaline phosphatase, Ca2+ATPase

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Authors: Homa Torabizadeh, Mohaddeseh Mikani

Abstract:

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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