Search results for: enzyme activity

Authors: Mustafa Erat

Abstract:

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

Abstract:

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

Abstract:

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: Seyed Azizollah Ghotb, Mohammad Chamani, Elmira Abdollahzadeh Esmaeili, Farhad Foroudi

Abstract:

the objective of this study is to measure the levels of cellulas activity of ostrich GI microorganisms, and comparing it with the levels of cellulas activity of rumen-s microorganisms, and also to estimate the probability of increasing enzyme activity with injecting different dosages (30%, 50% and 70%) of pure anaerobic goat rumen fungi. The experiment was conducted in laboratory and under a complete anaerobic condition (in vitro condition). 40 ml of “CaldWell" medium and 1.4g wheat straw were placed in incubator for an hour. The cellulase activity of ostrich microorganisms was compared with other treatments, and then different dosages (30%, 50% and 70%) of pure anaerobic goat rumen fungi were injected to ostrich microorganism-s media. Due to the results, cattle and goat with 2.13 and 2.08 I.U (international units) respectively showed the highest activity and ostrich with 0.91 (I.U) had the lowest cellulose activity (p < 0.05). Injecting 30% and 50% of anaerobic fungi had no significant incensement in enzyme activity, but with injecting 70% of rumen fungi to ostrich microorganisms culture a significant increase was observed 1.48 I.U. (p < 0.05).

Keywords: Cellulase enzyme, Microorganisms, Ostrich, Ruminants

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Authors: Justina I. R. Udotong

Abstract:

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

Abstract:

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

Abstract:

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: 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: 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: 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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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: Anna Yu. Muratova, Vera V. Kapitonova, Marina P. Chernyshova, Olga V. Turkovskaya

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This research was undertaken to study enzymatic activity in the shoots, roots, and rhizosphere of alfalfa (Medicago sativa L.) grown in quartz sand that was uncontaminated and contaminated with phenanthrene at concentrations of 10 and 100 mg kg-1. The higher concentration of phehanthrene had a distinct phytotoxic effect on alfalfa, inhibiting seed germination energy, plant survival, and biomass accumulation. The plant stress response to the environmental pollution was an increase in peroxidase activity. Peroxidases were the predominant enzymes in the alfalfa shoots and roots. The peroxidase profile in the shoots differed from that in the roots and had different isoenzyme numbers. 2,2'-Azinobis-(3-ethylbenzo-thiazoline-6-sulphonate) (ABTS) peroxidase was predominant in the shoots, and 2,7-diaminofluorene (2,2-DAF) peroxidase was predominant in the roots. Under the influence of phenanthrene, the activity of 2,7-DAF peroxidase increased in the shoots, and the activity of ABTS peroxidase increased in the roots. Alfalfa root peroxidases were the prevalent enzyme systems in the rhizosphere sand. Examination of the activity of alfalfa root peroxidase toward phenanthrene revealed the possibility of involvement of the plant enzyme in rhizosphere degradation of the PAH.

Keywords: Medicago sativa, enzymatic activity, peroxidase, phenanthrene.

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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: Surasak Siripornadulsil, Nutt Poomai, Wilailak Siripornadulsil

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Due to a high ethanol demand, the approach for  effective ethanol production is important and has been developed  rapidly worldwide. Several agricultural wastes are highly  abundant in celluloses and the effective cellulase enzymes do exist  widely among microorganisms. Accordingly, the cellulose  degradation using microbial cellulase to produce a low-cost substrate  for ethanol production has attracted more attention. In this  study, the cellulase producing bacterial strain has been isolated  from rich straw and identified by 16S rDNA sequence analysis as Acinetobacter sp. KKU44. This strain is able to grow and exhibit the cellulase activity. The optimal temperature for its growth and  cellulase production is 37°C. The optimal temperature of bacterial  cellulase activity is 60°C. The cellulase enzyme from  Acinetobacter sp. KKU44 is heat-tolerant enzyme. The bacterial culture of 36h. showed highest cellulase activity at 120U/mL when  grown in LB medium containing 2% (w/v). The capability of  Acinetobacter sp. KKU44 to grow in cellulosic agricultural wastes as a sole carbon source and exhibiting the high cellulase activity at high temperature suggested that this strain could be potentially developed further as a cellulose degrading strain for a production of low-cost substrate used in ethanol production. 

 

Keywords: Acinetobacter sp. KKU44, bagasse, cellulase enzyme, rice husk.

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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: 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: 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: 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: C. Zarcula, R. Croitoru, L. Corîci, C. Csunderlik, F. Peter

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Lipases are enzymes particularly amenable for immobilization by entrapment methods, as they can work equally well in aqueous or non-conventional media and long-time stability of enzyme activity and enantioselectivity is needed to elaborate more efficient bioprocesses. The improvement of Pseudomonas fluorescens (Amano AK) lipase characteristics was investigated by optimizing the immobilization procedure in hybrid organic-inorganic matrices using ionic liquids as additives. Ionic liquids containing a more hydrophobic alkyl group in the cationic moiety are beneficial for the activity of immobilized lipase. Silanes with alkyl- or aryl nonhydrolizable groups used as precursors in combination with tetramethoxysilane could generate composites with higher enantioselectivity compared to the native enzyme in acylation reactions of secondary alcohols. The optimal effect on both activity and enantioselectivity was achieved for the composite made from octyltrimethoxysilane and tetramethoxysilane at 1:1 molar ratio (60% increase of total activity following immobilization and enantiomeric ratio of 30). Ionic liquids also demonstrated valuable properties as reaction media for the studied reactions, comparable with the usual organic solvent, hexane.

Keywords: Ionic liquids, lipase, enantioselectivity, sol-gelimmobilization

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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: 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: Ratchadaporn Oonsivilai, Jutarat Manatwiyangkool, Anant Oonsivilai

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Theoptimal extraction condition of dried Phaseolus vulgaris powderwas studied. The three independent variables are raw material concentration, shaking and centrifugaltime. The dependent variables are both yield percentage of crude extract and alphaamylase enzyme inhibition activity. The experimental design was based on box-behnkendesign. Highest yield percentage of crude extract could get from extraction condition at concentration of 1, 0,1, concentration of 0.15 M ,extraction time for 2hour, and separationtime for60 min. Moreover, the crude extract with highest alpha-amylase enzyme inhibition activityoccurred by extraction condition at concentration of 0.10 M, extraction time for 2 min, and separation time for 45 min

Keywords: Extraction time, Optimal condition, Alpha-amylase enzymeinhibition activity

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Authors: A. Peter Amala Sujith, T.V. Hymavathi, P. Yasoda Devi

Abstract:

Lutein is a dietary oxycarotenoid which is found to reduce the risks of Age-related Macular Degeneration (AMD). Supercritical fluid extraction of lutein esters from marigold petals was carried out and was found to be much effective than conventional solvent extraction. The saponification of pre-concentrated lutein esters to produce free lutein was studied which showed a composition of about 88% total carotenoids (UV-VIS spectrophotometry) and 90.7% lutein (HPLC). The lipase catalyzed hydrolysis of lutein esters in conventional medium was investigated. The optimal temperature, pH, enzyme concentration and water activity were found to be 50°C, 7, 15% and 0.33 respectively and the activity loss of lipase was about 25% after 8 times re-use in at 50°C for 12 days. However, the lipase catalyzed hydrolysis of lutein esters in conventional media resulted in poor conversions (16.4%).

Keywords: lutein, preconcentration, saponification, lipase

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Authors: Katie Emerson, Sara Perez-Ojalvo, Jim Komorowski, Danielle Greenberg

Abstract:

The purpose of this study was to evaluate arginase activity levels in response to combinations of an inositol-stabilized arginine silicate (ASI; Nitrosigine®), L-arginine, and Inositol. Arginine acts as a vasodilator that promotes increased blood flow resulting in enhanced delivery of oxygen and nutrients to the brain and other tissues. Arginase, found in human serum, catalyzes the conversion of arginine to ornithine and urea, completing the last step in the urea cycle. Decreasing arginase levels maintains arginine and results in increased nitric oxide production. This study aimed to determine the most effective combination of ASI, L-arginine and inositol for minimizing arginase levels and therefore maximize ASI’s effect on cognition. Serum was taken from untreated healthy donors by separation from clotted factors. Arginase activity of serum in the presence or absence of test products was determined (QuantiChrom™, DARG-100, Bioassay Systems, Hayward CA). The remaining ultra-filtrated serum units were harvested and used as the source for the arginase enzyme. ASI alone or combined with varied levels of Inositol were tested as follows: ASI + inositol at 0.25 g, 0.5 g, 0.75 g, or 1.00 g. L-arginine was also tested as a positive control. All tests elicited changes in arginase activity demonstrating the efficacy of the method used. Adding L-arginine to serum from untreated subjects, with or without inositol only had a mild effect. Adding inositol at all levels reduced arginase activity. Adding 0.5 g to the standardized amount of ASI led to the lowest amount of arginase activity as compared to the 0.25 g, 0.75 g or 1.00g doses of inositol or to L-arginine alone. The outcome of this study demonstrates an interaction of the pairing of inositol with ASI on the activity of the enzyme arginase. We found that neither the maximum nor minimum amount of inositol tested in this study led to maximal arginase inhibition. Since the inhibition of arginase activity is desirable for product formulations looking to maintain arginine levels, the most effective amount of inositol was deemed preferred. Subsequent studies suggest this moderate level of inositol in combination with ASI leads to cognitive improvements including reaction time, executive function, and concentration.

Keywords: Arginine, blood flow, colorimetry, urea cycle.

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Authors: Ayuko Itsuki, Sachiyo Aburatani

Abstract:

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: 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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