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Sample records for plasma amylase activity

  1. Radiation effects on the parotid gland of mammals. Pt. 2. Modifications of plasma and parotid amylase activity

    Energy Technology Data Exchange (ETDEWEB)

    Balzi, M; Cremonini, D; Tomassi, I; Becciolini, A; Giannardi, G [Florence Univ. (Italy). Istituto di Radiologia; Pelu, G [I.N.R.C.A., Florence (Italy). Inst. of Radiology

    1979-08-01

    The early modifications of the activity of plasma and parotid amylase have been evaluated in rats after whole-body irradiation in different experimental conditions. The modifications observed in the rat are less evident than in humans. The results have shown an initial decrease and a subsequent increase appearing significant 74 hours after irradiation. This could be due to a direct action of ionizing radiation on the parotid glands. However we cannot exclude that the increase of ..cap alpha..-amylase could be determined by the intestinal radiation syndrome. In fact, this syndrome leads to a reduced food uptake, and consequently ..cap alpha..-amylase could accumulate in parotids.

  2. Plasma amylase activity as a biochemical indicator of radiation injury to salivary glands

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    Becciolini, A; Giannardi, G; Cionini, L; Porciani, S; Fallai, C; Pirtoli, L [Florence Univ. (Italy). Ist. di Radiologia

    1984-01-01

    Irradiation of the salivary glands produces a rapid increase of salivary amylase in serum, released by the highly radiation sensitive serous cells of the glands. Serial assays of salivary amylase in serum were performed in patients treated by radiation to the upper neck region. The changes observed were compared with the amount of salivary gland mass irradiated and with the dose fractionation modality used. The irradiated volume included either the entire salivary gland mass or less than 50 per cent of the gland. Two fractionation modalities were used: a conventional fractionation of 2 Gy per day, 5 times a week, or a multiple daily fractionation of 2 Gy, 3 times a day in two series of 4 days with a 4-day interval. Both parameters (salivary gland mass irradiated and fractionation modality used) significantly influenced the shape of the amylase curve in the serum. Serum amylase may therefore be considered a reliable biologic indicator of early injury to the salivary glands.

  3. Potency of Amylase-producing Bacteria and Optimization Amylase Activities

    Science.gov (United States)

    Indriati, G.; Megahati, R. R. P.; Rosba, E.

    2018-04-01

    Enzymes are capable to act as biocatalyst for a wide variety of chemical reactions. Amylase have potential biotechnological applications in a wide range of industrial processes and account for nearly 30% of the world’s enzyme market. Amylase are extracellular enzymes that catalyze the hydrolysis of internal α-1,4-glycosidic linkages in starch to dextrin, and other small carbohydrate molecules constituted of glucose units. Although enzymes are produced from animal and plant sources, the microbial sources are generally the most suitable for commercial applications. Bacteria from hot springs is widely used as a source of various enzymes, such as amylase. But the amount of amylase-producing bacteria is still very limited. Therefore it is necessary to search sources of amylase-producing bacteria new, such as from hot springs Pariangan. The purpose of this study was to isolation of amylase-producing bacteria from Pariangan hot spring, West Sumatera and amylase activity optimization. The results were obtained 12 isolates of thermophilic bacteria and 5 isolates of amyalse-producing bacteria with the largest amylolytic index of 3.38 mm. The highest amylase activity was obtained at 50°C and pH 7.5.

  4. Activity of secreted amylases in Aspergillus

    Energy Technology Data Exchange (ETDEWEB)

    Bocheva, S.S.; Kurnitskaya, L.N.

    1981-01-01

    When A. oryzae was cultivated in a synthetic liquid medium containing maltose as a sole source of C, the activity of extracellular amylase was 8.43-11.92 units/100 mL. Addition of 1.0% and 2.0% NaCl to the medium increased the amylase activity approximately 5- and 10-fold, respectively.

  5. Determination of amylase activity of crude extract from partially ...

    African Journals Online (AJOL)

    Amylase activity of crude extract from partially germinated mango seeds ( Mangifera oraphila) was determined using Caraway-Somogyi iodine/potassium iodide (IKI) method. The effects of varied pH and temperature were also investigated. The amylase was extracted with 0.1 M acetate buffer (pH 4.2). Amylase activity of the ...

  6. Factors influencing beta-amylase activity in sorghum malt

    CSIR Research Space (South Africa)

    Taylor, JRN

    1993-09-01

    Full Text Available isozyme of pI approximately 4.4-4.5, unlike the many isozymes all of higher pI in barley. However, like barley, sorghum beta-amylase was more temperature-labile than its alpha-amylase. Beta-amylase activity in sorghum malt was increased by germination time...

  7. Clinical Performance of a Salivary Amylase Activity Monitor During Hemodialysis Treatment

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    Masaru Shimazaki

    2008-01-01

    Full Text Available The hemodialysis procedure is thought to be a physical stressor in the majority of hemodialyzed patients. Previous studies suggest that elevated salivary amylase level may correlate with increased plasma norepinephrine level under psychological and physical stress conditions. In this study, we investigated biological stress reactivity during hemodialysis treatment using salivary amylase activity as a biomarker. Seven patients (male/female = 5/2, age:67.7+ /− 5.9 years who had been receiving regular 4 h hemodialysis were recruited. Salivary amylase activity was measured using a portable analyzer every hour during the hemodialysis session. Salivary amylase activity was shown to be relatively stable and constant throughout hemodialysis, whereas there were significant changes in systolic blood pressure and pulse rate associated with blood volume reduction. Our results show that hemodialysis treatment per se dose not affect salivary amylase activity.

  8. Optimisation of amylase and xylanase addition in dependance of white flour amylase activity

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    Lončar Davor M.

    2016-01-01

    Full Text Available In this study the effect of different quantities of added amylase to white wheat flours characterized with different activities of naturally existing amylases is tested. Response surface methodology is chosen to test the effects of main applied technological parameters on bread quality responses. Independent variables are chosen to be: quantity of added amylase and bulk fermentation time, while analysed responses are: specific volume, grain structure, bulk fermentation. Bread quality responses are statistically significant, while predicted and observed responses correspond very well, which allows good prediction of bread quality parameters based on applied technological parameters and flour characteristics. Score analysis shows that optimum quantity of amylase addition regarding bread quality depends on the activity of naturally existing amylases. Optimal quantity of added xylanase in bread samples made from both flour types is 0.004%. Xylanase improved properties of white wheat bread and higher effect is experienced with flour that has more active naturally existing amylases. Addition of amylase has statistically significantly increased a* values of crust. Addition of xylanase has statistically significantly decreased values of b* in comparison to the respective bread sample with only added amylase.

  9. The effect of carbohydrates on alpha-amylase activity measurements

    NARCIS (Netherlands)

    Baks, T.; Janssen, A.E.M.; Boom, R.M.

    2006-01-01

    The Ceralpha method can be used for ¿-amylase activity measurements during the hydrolysis of starch at high substrate concentrations (>40 wt.%). However, the results are affected by the carbohydrates present in the samples. The effect of carbohydrates on the Ceralpha ¿-amylase activity

  10. Activity and storage of commercial amylases in the 2013 Louisiana grinding season

    Science.gov (United States)

    A current problem in the application of amylases at sugarcane factories is the existence of a wide variation in the activities and activity per unit cost of commercial amylases. The efficiency of amylase action to break down starch in the factory is related to the activity of the amylase used. Until...

  11. Effect of radioactive isotope 32P upon alpha amylase activity and glucose concentration in chickens

    International Nuclear Information System (INIS)

    Kraljevic, P.; Emanovic, D.; Simpraga, M.; Nejedli, S.; Stojevic, Z.

    1996-01-01

    An attempt has been made to investigate whether alpha amylase activity and glucose concentration in blood plasma can serve as the help in establishing on early diagnosis of organic or functional damage caused by ionizing radiation in chickens. Fifty day old hybrid chickens of heavy 'Jata' breeds of both sexes, were treated by 32 P administered intramusculary as sodium orthophosphate in a single dose of 333 MBq per kilogram of body weight. Blood samples was taken from the wing vein on day 1, 3, 5, 7 and 10 after administration of 32 P. Alpha amylase activity and glucose concentration were determined spectrophotometrically using kits produced by 'Radonja', Sisak. Alpha amylase activity was decreased and glucose concentration was increased during investigated period. Yet, the further investigations are needed to find out whether these two parameters can be used for early diagnosis of injury in chicken organism by ionizing radiation. (author)

  12. THE DYNAMICS OF TOTAL AMYLASE\\'S ACTIVITY IN PANICUM MILIACEUM AND SETARIA GLAUCA DURING THE GERMINATION PERIOD

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    Elena Ciornea

    2006-08-01

    Full Text Available : It was studied the dynamics of total amylase\\'s activity in millet (Panicum miliaceum and bristle grass (Setaria glauca during the germination period. The enzymatic activity was determined by the Noelting - Brenfeld method, the results obtained being expressed in M maltose / g. In both millet and bristle grass, it was evidenced that both parameters taken into study (the species and the germination time do influence the enzymatic activity, although to a different extent.

  13. The relationship between the level of salivary alpha amylase activity and pain severity in patients with symptomatic irreversible pulpitis

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    Fatemeh Ahmadi-Motamayel

    2013-08-01

    Full Text Available Objectives Assessment of dental pain severity is very challenging in dentistry. Previous studies have suggested that elevated salivary alpha amylase may contribute to increased physical stresses. There is a close association between salivary alpha amylase and plasma norepinephrine under stressful physical conditions. The aim of this study was to evaluate the relationship between pain severity and salivary alpha amylase levels in patients with symptomatic irreversible pulpitis. Materials and Methods Thirty-six patients (20 females and 16 males with severe tooth pain due to symptomatic irreversible pulpitis were selected. The visual analogue scale (VAS score was used to assess the pain severity in each patient. Unstimulated whole saliva was collected, and the level of alpha amylase activity was assessed by the spectrophotometric method. Statistical analysis was performed using SPSS 13. Results The level of alpha amylase was significantly increased in the saliva in association with pain severity assessed by VAS. The salivary alpha amylase was also elevated with increased age and in males. Conclusions There was a significant correlation between the VAS pain scale and salivary alpha amylase level, which indicates this biomarker may be a good index for the objective assessment of pain intensity.

  14. Alternative method for quantification of alfa-amylase activity.

    Science.gov (United States)

    Farias, D F; Carvalho, A F U; Oliveira, C C; Sousa, N M; Rocha-Bezerrra, L C B; Ferreira, P M P; Lima, G P G; Hissa, D C

    2010-05-01

    A modification of the sensitive agar diffusion method was developed for macro-scale determination of alfa-amylase. The proposed modifications lower costs with the utilisation of starch as substrate and agar as supporting medium. Thus, a standard curve was built using alfa-amylase solution from Aspergillus oryzae, with concentrations ranging from 2.4 to 7,500 U.mL-1. Clear radial diffusion zones were measured after 4 hours of incubation at 20 A degrees C. A linear relationship between the logarithm of enzyme activities and the area of clear zones was obtained. The method was validated by testing alpha-amylase from barley at the concentrations of 2.4; 60; 300 and 1,500 U.mL-1. The proposed method turned out to be simpler, faster, less expensive and able to determine on a macro-scale alpha-amylase over a wide range (2.4 to 7,500 U.mL-1) in scientific investigation as well as in teaching laboratory activities.

  15. Alternative method for quantification of alfa-amylase activity

    Directory of Open Access Journals (Sweden)

    DF. Farias

    Full Text Available A modification of the sensitive agar diffusion method was developed for macro-scale determination of alfa-amylase. The proposed modifications lower costs with the utilisation of starch as substrate and agar as supporting medium. Thus, a standard curve was built using alfa-amylase solution from Aspergillus oryzae, with concentrations ranging from 2.4 to 7,500 U.mL-1. Clear radial diffusion zones were measured after 4 hours of incubation at 20 °C. A linear relationship between the logarithm of enzyme activities and the area of clear zones was obtained. The method was validated by testing α-amylase from barley at the concentrations of 2.4; 60; 300 and 1,500 U.mL-1. The proposed method turned out to be simpler, faster, less expensive and able to determine on a macro-scale α-amylase over a wide range (2.4 to 7,500 U.mL-1 in scientific investigation as well as in teaching laboratory activities.

  16. Salivary alpha-amylase: More than an enzyme Investigating confounders of stress-induced and basal amylase activity

    OpenAIRE

    Strahler, Jana

    2010-01-01

    Summary: Salivary alpha-amylase: More than an enzyme - Investigating confounders of stress-induced and basal amylase activity (Dipl.-Psych. Jana Strahler) The hypothalamus-pituitary-adrenal (HPA) axis and the autonomic nervous system (ANS) are two of the major systems playing a role in the adaptation of organisms to developmental changes that threaten homeostasis. The HPA system involves the secretion of glucocorticoids, including cortisol, into the circulatory system. Numerous studies hav...

  17. Characterization of the Activity and Stability of Amylase from Saliva and Detergent: Laboratory Practicals for Studying the Activity and Stability of Amylase from Saliva and Various Commercial Detergents

    Science.gov (United States)

    Valls, Cristina; Rojas, Cristina; Pujadas, Gerard; Garcia-Vallve, Santi; Mulero, Miquel

    2012-01-01

    This article presents two integrated laboratory exercises intended to show students the role of [alpha]-amylases (AAMYs) in saliva and detergents. These laboratory practicals are based on the determination of the enzymatic activity of amylase from saliva and different detergents using the Phadebas test (quantitative) and the Lugol test…

  18. Serum amylase and lipase activities after exploratory laparotomy in dogs.

    Science.gov (United States)

    Bellah, J R; Bell, G

    1989-09-01

    Serum amylase and lipase activities and creatinine concentration were determined before surgery, and at 1 and 2 days after exploratory laparotomy in 24 dogs. Examination of all viscera was done during each laparotomy, but a surgical procedure was not performed. The mean serum activities for lipase were: before surgery, 0.71 (0.0 to 2.0) Cherry Crandall units (CCU)/L; 1 day after surgery, 2.1 (0.0 to 4.5) CCU/L; and 2 days after surgery, 1.19 (0.0 to 3.9) CCU/L. The mean serum activities for amylase were: before surgery, 1,958 (1,027 to 3,426) IU/L; 1 day after surgery, 1,538 (937 to 2,659) IU/L; and 2 days after surgery, 1,663 (1,066 to 2,274) IU/L. Serum creatinine concentrations before surgery, 1 day after surgery, and 2 days after surgery were 0.88 (0.2 to 1.7) mg/dl, 0.78 (0.4 to 1.3) mg/dl, and 0.78 (0.3 to 1.3) mg/dl, respectively. Mean preoperative, day-1, and day-2 serum amylase activities and serum creatinine concentrations did not differ significantly from each other. Mean preoperative and day-2 serum lipase activities did not differ significantly; however, mean serum lipase activity was significantly greater when day 1 activities were compared with preoperative activities (P = 0.0002). Post-mortem examinations revealed no gross or histologic evidence of pancreatitis in any dog. The results of this study show that a 3 or more fold increase in serum lipase activity may occur after routine exploratory laparotomy in dogs without clinical signs or gross evidence of pancreatitis. Histologic evidence of pancreatitis was not found in the right pancreatic lobes in any dog.

  19. Heat activation and stability of amylases from Bacillus species

    African Journals Online (AJOL)

    Administrator

    2007-05-16

    May 16, 2007 ... as Bacillus macerans, Bacillus coagulans Bacillus licheniformis, Bacillus circulans, Bacillus megaterium, Bacillus polymyxa and Bacillus subtilis. Heat treatment at 70oC denatured the β-amylase component of the amylase source while α-amylase retained its potency at this temperature. Calcium.

  20. Salivary type alpha-amylase activity in serum and in urine of patients with lung adenocarcinoma

    International Nuclear Information System (INIS)

    Zakrzewska, I.; Wolska, K.; Koput, A.

    1993-01-01

    Total alpha-amylase activity in sera and urine of 30 patients with lung adenocarcinoma has been tested. The results were compared with control group of 30 healthy voluntaries. The activity of pancreatic type was differentiated from salivary alpha amylase. Salivary type was inhibited selectively by Triticum aestivum. Higher levels of total and salivary type amylase were noted in patients with lung adenocarcinoma in comparison to healthy control. The increase was significant (p<0.005). Correlation was observed between the activity of salivary type amylase and the stage of adenocarcinoma. (author)

  1. Effects of Pulsed Electric Field (PEF) Treatment on Enhancing Activity and Conformation of α-Amylase.

    Science.gov (United States)

    Tian, Mei-ling; Fang, Ting; Du, Mu-ying; Zhang, Fu-sheng

    2016-04-01

    To explore an efficient, safe, and speedy application of pulsed electric field (PEF) technology for enzymatic modification, effects of PEF treatment on the enzymatic activity, property and kinetic parameters of α-amylase were investigated. Conformational transitions were also studied with the aid of circular dichroism (CD) and fluorescence spectra. The maximum enzymatic activity of α-amylase was obtained under 15 kV/cm electric field intensity and 100 mL/min flow velocity PEF treatment, in which the enzymatic activity increased by 22.13 ± 1.14% compared with control. The activation effect could last for 18 h at 4 °C. PEF treatment could widen the range of optimum temperature for α-amylase, however, it barely exerted any effect on the optimum pH. On the other hand, α-amylase treated by PEF showed an increase of Vmax, t1/2 and ΔG, whereas a decrease of Km and k were observed. Furthermore, it can be observed from fluorescence and CD spectra that PEF treatment had increased the number of amino acid residues, especially that of tryptophan, on α-amylase surface with enhanced α-helices by 34.76% and decreased random coil by 12.04% on α-amylase when compared with that of untreated. These changes in structure had positive effect on enhancing α-amylase activity and property.

  2. Alpha-Amylase Activity in Blood Increases after Pharmacological, But Not Psychological, Activation of the Adrenergic System.

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    Urs M Nater

    Full Text Available Alpha-amylase in both blood and saliva has been used as a diagnostic parameter. While studies examining alpha-amylase activity in saliva have shown that it is sensitive to physiological and psychological challenge of the adrenergic system, no challenge studies have attempted to elucidate the role of the adrenergic system in alpha-amylase activity in blood. We set out to examine the impact of psychological and pharmacological challenge on alpha-amylase in blood in two separate studies.In study 1, healthy subjects were examined in a placebo-controlled, double-blind paradigm using yohimbine, an alpha2-adrenergic antagonist. In study 2, subjects were examined in a standardized rest-controlled psychosocial stress protocol. Alpha-amylase activity in blood was repeatedly measured in both studies.Results of study 1 showed that alpha-amylase in blood is subject to stronger increases after injection of yohimbine compared to placebo. In study 2, results showed that there was no significant effect of psychological stress compared to rest.Alpha-amylase in blood increases after pharmacological activation of the adrenergic pathways suggesting that sympathetic receptors are responsible for these changes. Psychological stress, however, does not seem to have an impact on alpha-amylase in blood. Our findings provide insight into the mechanisms underlying activity changes in alpha-amylase in blood in healthy individuals.

  3. Thermostable 𝜶-Amylase Activity from Thermophilic Bacteria Isolated from Bora Hot Spring, Central Sulawesi

    Science.gov (United States)

    Gazali, F. M.; Suwastika, I. N.

    2018-03-01

    α-Amylase is one of the most important enzyme in biotechnology field, especially in industrial application. Thermostability of α-Amylase produced by thermophilic bacteria improves industrial process of starch degradation in starch industry. The present study were concerned to the characterization of α-Amylase activity from indigenous thermophilic bacteria isolated from Bora hot spring, Central Sulawesi. There were 18 isolates which had successfully isolated from 90°C sediment samples of Bora hot spring and 13 of them showed amylolytic activity. The α-Amylase activity was measured qualitatively at starch agar and quantitatively based on DNS (3,5-Dinitrosalicylic acid) methods, using maltose as standard solution. Two isolates (out of 13 amylolytic bacteria), BR 002 and BR 015 showed amylolytic index of 0.8 mm and 0.5 mm respectively, after being incubated at 55°C in the 0.002% Starch Agar Medium. The α-Amylase activity was further characterized quantitatively which includes the optimum condition of pH and temperature of α-Amylase crude enzyme from each isolate. To our knowledge, this is the first report on isolation and characterization of a thermostable α-Amylase from thermophilic bacteria isolated from Central Sulawesi particularly from Bora hot spring.

  4. Copy number variation of human AMY1 is a minor contributor to variation in salivary amylase expression and activity.

    Science.gov (United States)

    Carpenter, Danielle; Mitchell, Laura M; Armour, John A L

    2017-02-20

    Salivary amylase in humans is encoded by the copy variable gene AMY1 in the amylase gene cluster on chromosome 1. Although the role of salivary amylase is well established, the consequences of the copy number variation (CNV) at AMY1 on salivary amylase protein production are less well understood. The amylase gene cluster is highly structured with a fundamental difference between odd and even AMY1 copy number haplotypes. In this study, we aimed to explore, in samples from 119 unrelated individuals, not only the effects of AMY1 CNV on salivary amylase protein expression and amylase enzyme activity but also whether there is any evidence for underlying difference between the common haplotypes containing odd numbers of AMY1 and even copy number haplotypes. AMY1 copy number was significantly correlated with the variation observed in salivary amylase production (11.7% of variance, P structure may affect expression, but this was not significant in our data.

  5. Amylase activity of a starch degrading bacteria isolated from soil ...

    African Journals Online (AJOL)

    STORAGESEVER

    2008-09-17

    Sep 17, 2008 ... Key words: Amylase, Bacillus, kitchen waste, starch degrading. INTRODUCTION ... several sources such as plant, animal and microbes. (Kathiresan .... Test. Response of the strain. Gram's staining. +. Shape. Bacilli. Motility. +.

  6. Effect of culture conditions on the activity of amylase used for alcoholic fermentation

    Energy Technology Data Exchange (ETDEWEB)

    Lee, S.D.; Ryu, Y.H.

    1973-01-01

    A wheat bran culture was used for media for the mold strains: Aspergillus oryzae, A. kawachii, A. usamii, and Rhizopus javanicus, to determine in which strain the amylase activity could be increased the most. The wheat bran media provided 47, 51, and 55% starch for each strain. To the media were added 3 nitrogen sources, viz.: (NH/sub 4/)/sub 2/SO/sub 4/, casein, and (NH/sub 4/)/sub 2/SO/sub 4/ - casein mixture. Each nitrogen source was made available at 2, 4, and 6% levels except only 2% (NH/sub 4/)/sub 2/SO/sub 4/ was used. The results obtained were as follows: (1) The ..cap alpha..-amylase activity was highest in media with 47% starch and 6% casein. (2) The ..beta..-amylase activity was highest in media with 51% starch and 2% (NH/sub 4/)/sub 2/SO/sub 4/ - casein. (3) Both ..cap alpha..- and ..beta..-amylase activities in A. usamii were highest in the media with 47% starch and no additional nitrogen source. (4) Of the 4 strains examined the ..cap alpha..- and ..beta..-amylase activities in R. javanicus were both relatively the highest. (5) The ..cap alpha..- and ..beta..-amylase activities of the strains examined decreased as the percentage of starch was increased except in R. javanicus.

  7. Alpha-amylase activity in wheat flour and breadmaking properties in relation to different climatic conditions

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    Rakita Slađana M.

    2015-01-01

    Full Text Available The aim of the present paper was to evaluate the influence of different climatic conditions on the activity of alpha-amylase in wheat samples and bread quality parameters. Three wheat varieties grown in three different localities in three years were chosen for this study. Commonly used methods for estimation of alpha-amylase activity in wheat grain were employed. The obtained results indicated that harvest year 2013, which was characterized with the excessive amount of rainfall, exhibited the highest level of alpha-amylase activity and the lowest values of the peak viscosity. The lowest alpha-amylase level and the highest peak viscosity and FN value were observed for samples harvested in 2012 which was characterized with the greatest number of days with an average daily temperature above 30 and 35°C. In addition, a decrease in Mixolab parameter torque C3 and specific bread loaf volume, as well as increase in the breakdown torque (C3-C4 of samples harvested in 2013 were observed, which could be attributed to rainy weather influencing increase in alpha-amylase activity. It is found that specific bread loaf volume of wheat samples is highest in 2012. Moreover, a negative correlation between alpha-amylase activity and specific bread volume for all the samples grown in three years was determined.

  8. Determination of amylase activity of crude extract from partially ...

    African Journals Online (AJOL)

    STORAGESEVER

    2009-07-20

    Jul 20, 2009 ... by its ability to gradually decrease the concentration of the starch solution used as substrate. The optimum ... synthesized during seed germination to mobilize stored food (starch .... basic property of all enzymes and is probably due to concomitant ... Use of potato starch as substrate for β-amylase by Bacillus ...

  9. Activity and cellular localization of amylases of rabbit cecal bacteria

    Czech Academy of Sciences Publication Activity Database

    Sirotek, Kamil; Marounek, Milan; Suchorská, O.

    2006-01-01

    Roč. 51, č. 4 (2006), s. 309-312 ISSN 0015-5632 R&D Projects: GA MŠk OC 848.001 Institutional research plan: CEZ:AV0Z50450515 Keywords : amylases Subject RIV: GH - Livestock Nutrition Impact factor: 0.963, year: 2006

  10. Cholecystokinin receptors: disparity between phosphoinositide breakdown and amylase releasing activity of CCK analogues in pancreas

    International Nuclear Information System (INIS)

    Lin, C.W.; Grant, D.; Bianchi, B.; Miller, T.; Witte, D.; Shue, Y.K.; Nadzan, A.

    1986-01-01

    Cholecystokinin (CCK) peptides are a family of hormones which also occur in brain. In pancreas CCK stimulates the release of amylase, a process that is dependent on the mobilization of intracellular Ca 2+ . Recent evidence suggests that inositol 1,4,5-trisphosphate, the breakdown product of phosphatidylinositol 4,5-bisphosphate, is responsible for the rise in intracellular Ca 2+ . Their laboratory has developed assays to study synthetic CCK analogues using radioligand binding, PI breakdown and amylase release. They have shown that there are good correlations among these three assay systems for the carboxy terminal fragments of CCK 8 . Recently, they have discovered synthetic analogues of CCK 4 that are full agonists in amylase release but are ineffective in causing PI breakdown. In particular, A-61576, Boc-5-amino-2-indolemethylene-pent-2-ene-1-oyl-Leu-Asp-Phe-NH 2 , is a full agonist in the amylase releasing assay, but is devoid of PI stimulating activity. A-61576 completely reverses the stimulation of PI response induced by CCK 8 , indicative of an antagonist. Since a mechanism other than the PI breakdown is responsible for amylase release by A-61576, they suggest that separate receptors are responsible for PI breakdown and amylase release

  11. Inhibitory Effect of Capparis spinosa Extract on Pancreatic Alpha-Amylase Activity

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    Mostafa Selfayan

    2016-04-01

    Full Text Available Background Diabetes mellitus is a metabolic disorder characterized by high blood glucose level caused due to deficiency of insulin secretion or insulin function. The inhibition of carbohydrate hydrolyzing enzymes such as α-amylase can be an important strategy for decrease postprandial blood glucose level in patients with type II diabetes. Plants contains different chemical constituents with potential for inhibition of α-amylase and hence maybe used as therapeutic. Objectives The aim of the present study is to investigate the effect of the ethanolic extract of Capparis spinosa on pancreatic α-amylase activities to find out the relevance of the plant in controlling blood sugar. Materials and Methods In this experimental study, root and leaves of C. spinosa were tested for α-amylase inhibition. Different concentrations (1.56, 3.12, 6.25, 12.5 and 25 mg/mL of extracts were incubated with enzyme substrate solution and the spectrometric method used for measure enzyme activity. Also acarbose was used as the standard inhibitor. Results Both root and leaves extracts showed inhibition of α-amylase (root = 97.31% and leaves = 98.92%. The root and leaves extracts of C. spinosa exhibited appreciable α-amylase inhibitory activity with an IC50 values 5.93 mg/mL and 3.89 mg/mL respectively, when compared with acarbose (IC50 value 0.038 mg/mL. Conclusions This study supports that root and leaves extracts of C. spinosa exhibit considerable α-amylase inhibitory activities. These results could be useful for developing functional foods by combination of plant-based foods for treatment of diabetes mellitus.

  12. IN VITRO ANTIOXIDANT AND α-AMYLASE INHIBITION ACTIVITIES OF PANCHSAKAR CHURNA

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    Ashok Kumar B.S.

    2013-12-01

    Full Text Available Panchsakar Churna is the composition of Cassia angustifolia, Terminalia chebula, Zingiber officinale, Foeniculum vulgare and Saindhava lavana. Aqueous extract of churna was used to investigate antioxidant activity by ferrous ion chelating assay and ferric reducing power and alpha amylase inhibition activity by dinitrosalicylic acid method (DNSA. Aqueous extract of churna showed maximum ferrous chelating activity - 42.01 and ferric reducing power - 1.5 and 83.33 % of inhibition protein denaturation at 1000 µg/ml. Panchsakar churna showed significant antioxidant and alpha amylase inhibition activities.

  13. Salinity Inhibits Rice Seed Germination by Reducing α-Amylase Activity via Decreased Bioactive Gibberellin Content

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    Li Liu

    2018-03-01

    Full Text Available Seed germination plays important roles in the establishment of seedlings and their subsequent growth; however, seed germination is inhibited by salinity, and the inhibitory mechanism remains elusive. Our results indicate that NaCl treatment inhibits rice seed germination by decreasing the contents of bioactive gibberellins (GAs, such as GA1 and GA4, and that this inhibition can be rescued by exogenous bioactive GA application. To explore the mechanism of bioactive GA deficiency, the effect of NaCl on GA metabolic gene expression was investigated, revealing that expression of both GA biosynthetic genes and GA-inactivated genes was up-regulated by NaCl treatment. These results suggest that NaCl-induced bioactive GA deficiency is caused by up-regulated expression of GA-inactivated genes, and the up-regulated expression of GA biosynthetic genes might be a consequence of negative feedback regulation of the bioactive GA deficiency. Moreover, we provide evidence that NaCl-induced bioactive GA deficiency inhibits rice seed germination by decreasing α-amylase activity via down-regulation of α-amylase gene expression. Additionally, exogenous bioactive GA rescues NaCl-inhibited seed germination by enhancing α-amylase activity. Thus, NaCl treatment reduces bioactive GA content through promotion of bioactive GA inactivation, which in turn inhibits rice seed germination by decreasing α-amylase activity via down-regulation of α-amylase gene expression.

  14. Traditionally used plants in diabetes therapy: phytotherapeutics as inhibitors of alpha-amylase activity

    Directory of Open Access Journals (Sweden)

    Ingrid Funke

    Full Text Available Diabetes mellitus is a metabolic disorder characterized by chronic hyperglycaemia. There are many and diverse therapeutic strategies in the management of Type 2 diabetes. The inhibition of alpha-amylase activity is only one possibility to lower postprandial blood glucose levels. In our in-vitro studies we could demonstrate that different plants, mostly traditionally used in common diabetic therapy in Africa or Europe, are able to inhibit alpha-amylase, which is responsible for the breakdown of oligosaccharides into monosaccharides which are absorbed. An inhibition of alpha-amylase activity of 90% was seen with the extract of the leaves of Tamarindus indica. To quantify inhibtion rates, acarbose was used (IC50: 23.2 µM. Highest inhibition level of acarbose in our testmodel was about 85%. Additionally tests with pure polyphenolic compounds might explain the biological activity of the selected plants.

  15. Amylase Zymography.

    Science.gov (United States)

    Andrades, Adarelys; Contreras, Lellys M

    2017-01-01

    Amylase zymography was carried out for the detection of amylases produced by a Geobacillus stearothermophilus strain isolated from the Thermal Center "Las Trincheras" in Venezuela. Zymography is an electrophoretic technique used to study hydrolases by means of thin gels containing copolymerized-specific substrates, under nonreducing conditions. In this study, 0.1% starch was incorporated into the gel as substrate. The formation of clear zones against a dark background in the gel stained with iodine indicated the presence of amylolytic activity. The thermophilic bacteria released several extracellular amylases to a selective growth medium supplemented with 1% soluble starch at 55 °C after 40 h incubation. The amylolytic enzymes showed an optimum temperature of 60 °C and an optimum pH at 6.0. The amylases were partially purified by cold acetone precipitation followed by two chromatographic techniques. These purified amylases showed different molecular masses which were determined by sodium dodecyl sulfate gel electrophoresis and confirmed by zymography.

  16. Smart phone: a popular device supports amylase activity assay in fisheries research.

    Science.gov (United States)

    Thongprajukaew, Karun; Choodum, Aree; Sa-E, Barunee; Hayee, Ummah

    2014-11-15

    Colourimetric determinations of amylase activity were developed based on a standard dinitrosalicylic acid (DNS) staining method, using maltose as the analyte. Intensities and absorbances of red, green and blue (RGB) were obtained with iPhone imaging and Adobe Photoshop image analysis. Correlation of green and analyte concentrations was highly significant, and the accuracy of the developed method was excellent in analytical performance. The common iPhone has sufficient imaging ability for accurate quantification of maltose concentrations. Detection limits, sensitivity and linearity were comparable to a spectrophotometric method, but provided better inter-day precision. In quantifying amylase specific activity from a commercial source (P>0.02) and fish samples (P>0.05), differences compared with spectrophotometric measurements were not significant. We have demonstrated that iPhone imaging with image analysis in Adobe Photoshop has potential for field and laboratory studies of amylase. Copyright © 2014 Elsevier Ltd. All rights reserved.

  17. A Kinetic Model to Explain the Maximum in alpha-Amylase Activity Measurements in the Presence of Small Carbohydrates

    NARCIS (Netherlands)

    Baks, T.; Janssen, A.E.M.; Boom, R.M.

    2006-01-01

    The effect of the presence of several small carbohydrates on the measurement of the -amylase activity was determined over a broad concentration range. At low carbohydrate concentrations, a distinct maximum in the -amylase activity versus concentration curves was observed in several cases. At higher

  18. ON THE ACTIVITY OF α-AMYLASE IN SOME CULTURE AND SPONTANEOUS GRAMINACEAE, DURING THEIR GERMINATION PERIOD

    Directory of Open Access Journals (Sweden)

    Elena Ciornea

    2006-08-01

    amylase (595.871 M maltose / g has been evidenced 12 hours after the beginning of germination, while, in the case of bristle grass, -amylase attains the maximum level of its activity after 144 hours of germination (429.824 M maltose / g.

  19. Halotolerant ability and α-amylase activity of some saltwater fungal isolates

    NARCIS (Netherlands)

    Niknejad, F.; Moshfegh, M.; Najafzadeh, M.J.; Houbraken, J.; Rezaei, S.; Zarrini, G.; Faramarzi, M.A.; Nafissi-Varcheh, N.

    2013-01-01

    Four halotolerant fungal isolates originating from the saltwater Lake Urmia in Iran were selected during a screening program for salt resistance and α-amylase activity. The isolates were identified based on sequencing the ITS region and a part of the β-tubulin gene, as Penicillium chrysogenum

  20. Changes in the composition of the pulp, alpha-amylase activity and ...

    African Journals Online (AJOL)

    The laboratory rotting of Egusi Fruits was completed in 120 hours. At this stage pulp became soft and the seeds were extracted easily with the fingers. The changes in the composition of the pulp, alpha-amylase activity and titratable acidity during the controlled rotting of egusi fruit (Colocynthis citrullus L.) for the harvesting of ...

  1. Enhancing Maritime Education and Training: Measuring a Ship Navigator's Stress Based on Salivary Amylase Activity

    Science.gov (United States)

    Murai, Koji; Wakida, Shin-Ichi; Miyado, Takashi; Fukushi, Keiichi; Hayashi, Yuji; Stone, Laurie C.

    2009-01-01

    Purpose: The purpose of this paper is to propose that the measurement of salivary amylase activity is an effective index to evaluate the stress of a ship navigator for safe navigation training and education. Design/methodology/approach: Evaluation comes from the simulator and actual on-board experiments. The subjects are real captains who have…

  2. In vitro antioxidant and α-amylase inhibition activities of spiced red ...

    African Journals Online (AJOL)

    Spiced chili paste (green or red), locally known as Datta, is a traditional popular spicy paste consumed in Ethiopia. This study investigated the total phenolic contents (TPC), total flavonoid contents (TFC), in vitro antioxidant, and α-amylase inhibition activities of water, acetone, petroleum ether, methanol, and 80% methanol ...

  3. Partial characterization of cold active amylases and proteases of Streptomyces sp. from Antarctica

    Directory of Open Access Journals (Sweden)

    Mihaela Cotârleţ

    2011-09-01

    Full Text Available The aim of this study was to isolate novel enzyme-producing bacteria from vegetation samples from East Antarctica and also to characterize them genetically and biochemically in order to establish their phylogeny. The ability to grow at low temperature and to produce amylases and proteases cold-active was also tested. The results of the 16S rRNA gene sequence analysis showed that the 4 Alga rRNA was 100% identical to the sequences of Streptomyces sp. rRNA from Norway and from the Solomon Islands. The Streptomyces grew well in submerged system at 20ºC, cells multiplication up to stationary phase being drastically increased after 120 h of submerged cultivation. The beta-amylase production reached a maximum peak after seven days, while alpha-amylase and proteases were performing biosynthesis after nine days of submerged cultivation at 20ºC. Newly Streptomyces were able to produce amylase and proteases in a cold environment. The ability to adapt to low temperature of these enzymes could make them valuable ingredients for detergents, the food industry and bioremediation processes which require low temperatures.

  4. Effect of the glass transition temperature on alpha-amylase activity in a starch matrix.

    Science.gov (United States)

    Chaudhary, Vinita; Panyoyai, Naksit; Small, Darryl M; Shanks, Robert A; Kasapis, Stefan

    2017-02-10

    This study optimises a protocol for the estimation of α-amylase activity in a condensed starch matrix in the vicinity of the glass transition region. Enzymatic activity on the vitrified starch system was compared with that of a reference substrate, maltodextrin. The activity was assayed as the rate of release of reducing sugar using a dinitrosalicylic acid procedure. The condensed carbohydrate matrices served the dual purpose of acting as a substrate as well as producing a pronounced effect on the ability to enzymatic hydrolysis. Activation energies were estimated throughout the glass transition region of condensed carbohydrate preparations based on the concept of the spectroscopic shift factor. Results were used to demonstrate a considerable moderation by the mechanical glass transition temperature, beyond the expected linear effect of the temperature dependence, on the reaction rate of starch hydrolysis by α-amylase in comparison with the low-molecular weight chain of maltodextrin. Copyright © 2016. Published by Elsevier Ltd.

  5. [Activity of alpha-amylase and concentration of protein in saliva of pregnant women].

    Science.gov (United States)

    Ciejak, Magdalena; Olszewska, Maria; Jakubowska, Katarzyna; Zebiełowicz, Dariusz; Safranow, Krzysztof; Chlubek, Dariusz

    2007-01-01

    One of the hypothetical reasons of the increased incidence of caries in women during the pregnancy may be the increased activity of alpha-amylase, which can be found in their saliva. The enzyme takes part in the process of decomposition of simple sugars, which make basic substrate for caries-causing bacteria. The aim of the paper was the evaluation of the influence of pregnancy and gestational age on the activity of alpha-amylase and the concentration of protein in women's saliva. The examined group consisted of 64 pregnant women at age 17-39, between 21st and 40th week of pregnancy. The control group consisted of 44 healthy women at age 20-35, who were not pregnant. In saliva, which was taken before morning meal, without stimulation, protein concentration was determined by Bradford method and the activity of amylase was determined by kinetic method. The activity of amylase correlated strongly and positively with protein concentration in saliva of both the pregnant (RS = +0.65; p women. There were no significant differences between examined parameters in the examined and the control group. It has been observed in the examined group, that there is the significant negative correlation between protein concentration in saliva and the week of pregnancy (RS = -0.35; p increased caries incidence of pregnant women. However, the observed changes of total protein concentration in saliva during pregnancy, suggest that the exact cognition of proteins in pregnant women's saliva may reveal new mechanisms, which lead to an increase of caries risk.

  6. Chemical modification, antioxidant and α-amylase inhibitory activities of corn silk polysaccharides.

    Science.gov (United States)

    Chen, Shuhan; Chen, Haixia; Tian, Jingge; Wang, Yanwei; Xing, Lisha; Wang, Jia

    2013-10-15

    Water-soluble corn silk polysaccharides (CSPS) were chemically modified to obtain their sulfated, acetylated and carboxymethylated derivatives. Chemical characterization and bioactivities of CSPS and its derivatives were comparatively investigated by chemical methods, gas chromatography, gel filtration chromatography, scanning electron microscope, infrared spectroscopy and circular dichroism spectroscopy, scavenging DPPH free radical assay, scavenging hydroxyl radical assay, ferric reducing power assay, lipid peroxidation inhibition assay and α-amylase activity inhibitory assay, respectively. Among the three derivatives, carboxylmethylated polysaccharide (C-CSPS) demonstrated higher solubility, narrower molecular weight distribution, lower intrinsic viscosity, a hyperbranched conformation, significantly higher antioxidant and α-amylase inhibitory abilities compared with the native polysaccharide and other derivatives. C-CSPS might be used as a novel nutraceutical agent for human consumption. Copyright © 2013 Elsevier Ltd. All rights reserved.

  7. Inhibitory effects of edible seaweeds, polyphenolics and alginates on the activities of porcine pancreatic α-amylase

    DEFF Research Database (Denmark)

    Zaharudin, Nazikussabah Binti; Asunción Salmeán, Armando; Dragsted, Lars Ove

    2018-01-01

    Edible seaweeds are valuable because of their organoleptic properties and complex polysaccharide content. A study was conducted to investigate the potential of dried edible seaweed extracts, its potential phenolic compounds and alginates for α-amylase inhibitory effects. The kinetics of inhibition......,5-dihydroxybenzoic acid was found to be a potent inhibitor of α-amylase with an IC50 value of 0.046 ± 0.004 mg/ml. Alginates found in brown seaweeds appeared to be potent inhibitors of α-amylase activity with an IC50 of (0.075 ± 0.010–0.103 ± 0.017) mg/ml, also a mixed-type inhibition. Overall, the findings provide...... information that crude extracts of brown edible seaweeds, phenolic compounds and alginates are potent α-amylase inhibitors, thereby potentially retarding glucose liberation from starches and alleviation of postprandial hyperglycaemia....

  8. Chloride Activated Halophilic α-Amylase from Marinobacter sp. EMB8: Production Optimization and Nanoimmobilization for Efficient Starch Hydrolysis

    Directory of Open Access Journals (Sweden)

    Sumit Kumar

    2015-01-01

    Full Text Available Halophiles have been perceived as potential source of novel enzymes in recent years. The interest emanates from their ability to catalyze efficiently under high salt and organic solvents. Present work encompasses production optimization and nanoimmobilization of an α-amylase from moderately halophilic Marinobacter sp. EMB8. Media ingredients and culture conditions were optimized by “one-at-a-time approach.” Starch was found to be the best carbon source at 5% (w/v concentration. Glucose acted as catabolic repressor for amylase production. Salt proved critical for amylase production and maximum production was attained at 5% (w/v NaCl. Optimization of various culture parameters resulted in 48.0 IU/mL amylase production, a 12-fold increase over that of unoptimized condition (4.0 IU/mL. α-Amylase was immobilized on 3-aminopropyl functionalized silica nanoparticles using glutaraldehyde as cross-linking agent. Optimization of various parameters resulted in 96% immobilization efficiency. Starch hydrolyzing efficiency of immobilized enzyme was comparatively better. Immobilized α-amylase retained 75% of its activity after 5th cycle of repeated use.

  9. Heat activation and stability of amylases from Bacillus species | Ajayi ...

    African Journals Online (AJOL)

    Leitch and Collier sporulating Bacillus medium was used to isolate some strains of Bacillus species from soil, wastewater and food sources in Ibadan, Oyo State, Nigeria, by heat activation method. Heat treatment at 80oC allowed the growth of sporulating Bacillus species, in the culture sample source without other bacteria ...

  10. Activities of amylase, trypsin and chymotrypsin of pancreas and ...

    African Journals Online (AJOL)

    The digestive enzyme activities of the pancreas and small intestinal segments were examined in two breeds of chickens that differ in growth rate over the period of 1 day (1-d) to 4-months (120-d) of age. The total body weight (BW) of the red jungle fowl (RJF) increased slowly during the experiment, in contrast to the ...

  11. Aqueous extracts of Roselle (Hibiscus sabdariffa Linn.) varieties inhibit α-amylase and α-glucosidase activities in vitro.

    Science.gov (United States)

    Ademiluyi, Adedayo O; Oboh, Ganiyu

    2013-01-01

    This study sought to investigate the inhibitory effect of aqueous extracts of two varieties (red and white) of Hibiscus sabdariffa (Roselle) calyces on carbohydrate hydrolyzing enzymes (α-amylase and α-glucosidase), with the aim of providing the possible mechanism for their antidiabetes properties. Aqueous extracts were prepared (1:100 w/v) and the supernatant used for the analysis. The extracts caused inhibition of α-amylase and α-glucosidase activities in vitro.The IC(50) revealed that the red variety (25.2 μg/mL) exhibited higher α-glucosidase inhibitory activity than the white variety (47.4 μg/mL), while the white variety (90.5 μg/mL) exhibited higher α-amylase inhibitory activity than the red variety (187.9 μg/mL). However, the α-glucosidase inhibitory activities of both calyces were higher than that of their α-amylase. In addition, the red variety possessed higher antioxidant capacity as exemplified by the (•)OH scavenging abilities, Fe(2+) chelating ability, and inhibition of Fe(2+)-induced pancreatic lipid peroxidation in vitro. The enzyme inhibitory activities and antioxidant properties of the roselle extracts agreed with their phenolic content. Hence, inhibition of α-amylase and α-glucosidase, coupled with strong antioxidant properties could be the possible underlying mechanism for the antidiabetes properties of H. sabdariffa calyces; however, the red variety appeared to be more potent.

  12. Modification of the activity of an alpha-amylase from Bacillus licheniformis by several surfactants

    OpenAIRE

    Bravo Rodríguez,Vicente; Jurado Alameda,Encarnación; Martínez Gallegos,Juan Francisco; Reyes Requena,Antonia; García López,Ana Isabel; Sampaio Cabral,Joaquim Manuel; Fernandes,Pedro; Pina da Fonseca,Luis Joaquim

    2006-01-01

    The influence of different commercial surfactants on the enzymatic activity of a commercial alpha-amylase from Bacillus licheniformis (Termamyl 300 L) has been studied. As non-ionic surfactants, alkyl polyglycosides (Glucopon® 215, Glucopon® 600 and Glucopon® 650) were studied, as were fatty alcohol ethoxylates (Findet 1214N/23 and Findet 10/15), and nonyl phenol ethoxylate (Findet 9Q/21.5NF). Also, an anionic surfactant, linear alkyl benzene sulfonate (LAS) was assayed. In general, none of t...

  13. Effects of Cadmium Stress on Seed Germination, Seedling Growth and Seed Amylase Activities in Rice (Oryza sativa

    Directory of Open Access Journals (Sweden)

    Jun-yu HE

    2008-12-01

    Full Text Available Two rice varieties, Xiushui 110 with high cadmium (Cd tolerance and Xiushui 11 with low Cd tolerance were used to study the effects of Cd stress on seed germination, seedling growth and amylase activities. The low cadmium concentration had little effect on seed germination rate. However, cadmium stress could significantly inhibit plumule and radicle growth, especially for radicle growth. Germination index, vigour index, radicle length and amylase activities of Xiushui 11 decreased more significantly with the increasing cadmium level compared with Xiushui 110. The cadmium content in seedlings of Xiushui 11 was higher than that in Xiushui 110 when the cadmium concentration exceeded 5 μmol/L, which caused lower mitotic index in root tips and amylase activities, and more serious cadmium toxicity in Xiushui 11.

  14. α-/β-Glucosidase and α-Amylase Inhibitory Activities of Roselle (Hibiscus sabdariffa L. Ethanol Extract

    Directory of Open Access Journals (Sweden)

    Marisca Evalina Gondokesumo

    2017-03-01

    Full Text Available Background: Diabetes mellitus is a metabolic disease, characterized by hyperglycemia due to disturbance in both insulin secretion and function. One of theurapeutic approaches is to reduce blood glucose levels by inhbiting α-/β-glucosidase and α-amylase involved in carbohydrate digestion. Thus, inhibition of these enzymes play important role in the treatment of diabetes mellitus. Roselle (Hibiscus sabdariffa L. has been known to have several medicinal properties and potency as an antidiabetics agents. This reseacrh aimed to observe antidiabetic properties of roselle ethanol extract (REE towards α-glucosidase, β-glucosidase and α-amylase. Materials and Methods: REE was done with maceration technique using diluent of 70% ethanol. Antidiabetic properties were measured by inhibitory activity of α-amylase, α-glucosidase and β-glucosidase. Results: REE was able to inhibit α-/β-glucosidase and α-amylase in the highest concentration with inhibition percentage of 72.68, 47.34 and 73.08% respectively, and were comparable with Acarbose of 81.49, 50.97, 73.08%. The median inhibitory concentration (IC50 of α-/β-glucosidase and α-amylase of REE were 15.81, 41.77, 18.09 μg/mL respectively, and Acarbose were 9.45, 22.57, 3.64 μg/mL respectively. Conclusions: REE inhibits α-/β-glucosidase and α-amylase. Keywords: Roselle, Acarbose, α-glucosidase, β-glucosidase, α-amylase, antidiabetic

  15. Nutrient Content, Phytonutrient Composition, Alpha Amylase, Alpha Glucosidase Inhibition Activity and Antioxidant Activity of the Stoechospermum Marginatum Collected in Pre Monsoon Season

    OpenAIRE

    Reka Palanivel; Thahira Banu Azeez; Seethalakshmi Muthaya

    2017-01-01

    The objective of this study was to investigate the nutrient content, phytonutrient composition, physicochemical properties, alpha amylase and alpha glucosidase inhibition activity and antioxidant activity of the brown algae Stoechospermum marginatum collected from Gulf of Mannar, Tamil Nadu, India in pre monsoon season (June- September, 2015). Six and eight hours of ethanol and aqueous extract of Stoechospermum marginatum were used for phytonutrient screening, alpha amylase, alpha glucosidase...

  16. In-vitro alpha amylase inhibitory activity of the leaf extracts of Adenanthera pavonina.

    Science.gov (United States)

    Wickramaratne, M Nirmali; Punchihewa, J C; Wickramaratne, D B M

    2016-11-15

    Diabetes has caused a major burden to the health sector in the developing countries and has shown an increasing trend among the urban population. It is estimated that most patients are with type II diabetes which could be easily treated with dietary changes, exercise, and medication. Sri Lanka carries a long history ayurvedic medicine where it uses the plant for treating many diseases. Therefore it is important to screen medicinal plants scientifically so they could be used safely and effectively in the traditional medical system and also be used for further investigations. Adenanthera pavonina is a plant used in the Ayurvedic medical system in Sri Lanka for treating many diseases including diabetics. We evaluated the anti-diabetic properties and the antioxidant properties of Adenanthera pavonina leaves. The methanol extract of the leaves was sequentially extracted with petroleum ether and thereafter was partitioned between EtOAc, and water. The α-amylase inhibition assay was performed using the 3,5- dinitrosalicylic acid method. The antioxidant activities were measured using the DPPH free radical scavenging activity and the total phenolic content using Folin-Ciocalteu's reagent. The cytotoxicity of the extract was evaluated using the Brine shrimp bioassay. The IC 50 values of α amylase inhibitory activity of MeOH, EtOAc, petroleum ether, and water were 16.16 ± 2.23, 59.93 ± 0.25, 145.49 ± 4.86 and 214.85 ± 9.72 μg/ml respectively and was similar to that of Acarbose (18.63 ± 1.21 (μg/ml). Antioxidant activities were also determined and the EtOAc fraction showed the highest total phenolic content (34. 62 ± 1.14 mg/g extract) and the highest DPPH scavenging activity with an IC 50 of 249.92 ± 3.35 μg/ml. The leaf extracts of Adenanthera pavonina exhibit remarkable α-amylase inhibitory activity in the crude methanolic extract. Hence leaves of Adenanthera pavonina has a potential to be used as a regular green vegetable and

  17. Irradiation Effect on Secreting Function, Amylase Activity and Nucleic Acid Contents of Rat Parotid Gland

    International Nuclear Information System (INIS)

    Cho, Young Jin; Park, Tae Won

    1990-01-01

    This experiment was performed to clarify the effects of 60 Co gamma irradiation on secretory function, amylase activity and contents of nucleic acids of parotid gland in rat. Experimental animals were divided into 6th hours, 3rd, 7th, 14th and 28th days after irradiation and control. The experimental animals are singly irradiated with 20 Gy (2,000 rad) through protective lead block. Secretory function of parotid gland was evaluated by uptake and clearance of 99m TcO 4 . 99m TcO 4 , 0.2μ ci/gm, was injected into peritonium in uptake groups. Rats were sacrificed with cervical dislocation after 30 minutes and gland was excised. In the clearance group, pilocarpine nitrate (8 mg/kg) was intraperitoneally injected at 30 minutes after 99m TcO 4 injection and rats were sacrificed 30 minutes after pilocarpine infection. Radioactivity of excised parotid gland was measured by using of gamma counter and stimulation-secretion coefficient, uptake radioactivity divided by clearance radioactivity, was calculated. Amylase activity and contents of DNA and RNA were determined by spectrophotometry. The results obtained were as follows: 1. In the uptake test, the radioactivity of 99m TcO 4 per unit weight increase in experimental group except 6th hours group, compared with control groups and showed a peak at 3rd days after irradiation. 2. In the clearance test, the radioactivity of 99m TcO 4 per unit weight rose to a peak at 3rd days after irradiation and gradually recovered thereafter. 3. Stimulation-secretion coefficient of parotid gland decreased at 6th hours, 3rd and 7th days after irradiation, and gradually increased. 4. Amylase activity of parotid gland decreased in 3rd and 7th days group, and especially lowest in 3rd days after irradiation. 5. The contents of DNA showed no definite difference in all the experimental groups, but RNA was seemed to decrease with time after irradiation.

  18. Effect of neohesperidin dihydrochalcone on the activity and stability of alpha-amylase: a comparative study on bacterial, fungal, and mammalian enzymes.

    Science.gov (United States)

    Kashani-Amin, Elaheh; Ebrahim-Habibi, Azadeh; Larijani, Bagher; Moosavi-Movahedi, Ali Akbar

    2015-10-01

    Neohesperidin dihydrochalcone (NHDC) was recently introduced as an activator of mammalian alpha-amylase. In the current study, the effect of NHDC has been investigated on bacterial and fungal alpha-amylases. Enzyme assays and kinetic analysis demonstrated the capability of NHDC to significantly activate both tested alpha-amylases. The ligand activation pattern was found to be more similar between the fungal and mammalian enzyme in comparison with the bacterial one. Further, thermostability experiments indicated a stability increase in the presence of NHDC for the bacterial enzyme. In silico (docking) test locates a putative binding site for NHDC on alpha-amylase surface in domain B. This domain shows differences in various alpha-amylase types, and the different behavior of the ligand toward the studied enzymes may be attributed to this fact. Copyright © 2015 John Wiley & Sons, Ltd.

  19. Relationships between falling number, a-Amylase activity, milling, and sponge cake quality of soft white wheat

    Science.gov (United States)

    Falling Number of wheat is an important quality predictor and carries with it significant economic impact. Lower Falling Numbers are associated with higher a-amylase activity and poorer soft wheat end-use quality, especially sponge cake. In the present study two sample sets were examined, the first ...

  20. alfa-Amylase as a reliable and convenient measure of sympathetic activity: don’t start salivating just yet!

    NARCIS (Netherlands)

    Bosch, JA; Veerman, J.W.; de Geus, E.J.C.; Proctor, J.

    2011-01-01

    Recent years have seen a growing interest in salivary α-amylase (sAA) as a non-invasive marker for sympathetic nervous system (SNS) activity. Saliva offers many advantages as a biomarker fluid and sAA is one of its most plentiful components. sAA is a digestive enzyme that breaks down starch, which

  1. Effects of exogenous gamma-aminobutyric acid on α-amylase activity in the aleurone of barley seeds.

    Science.gov (United States)

    Sheng, Yidi; Xiao, Huiyuan; Guo, Chunli; Wu, Hong; Wang, Xiaojing

    2018-03-03

    Gamma-aminobutyric acid (GABA), a nonprotein amino acid, often accumulates in plants exposed to certain environmental stimuli. Previous studies indicated that a closed relationship existed between endogenous GABA and seed germination. However, there are few studies on the effect of exogenous GABA on seed germination. The objective of this study was to explore whether exogenous GABA affected α-amylase activity which the activation is an important stage in seed germination. The level of endogenous GABA in barley seeds rose gradually during germination, suggesting that endogenous GABA was involved in germination. We measured starch degradation under application of various concentration GABA and found that GABA promoted seed starch degradation with a dose-responsive effect. The relationship between GABA and α-amylase activity was investigated by treating barley aleurone with exogenous GABA. The result showed that α-amylase activity began to rise after about 24 h and reached a peak at 48 h. Molecular evidence suggested that GABA increased α-amylase gene expression. We explore the possible roles played by GABA in signal transduction. Copyright © 2018 Elsevier Masson SAS. All rights reserved.

  2. Enzymatic activity of α-amylase in alimentary tract Spodoptera littoralis (Boisduval (Lepidoptera: Noctuidae: Characterization and Compartmentalization

    Directory of Open Access Journals (Sweden)

    Ali Darvishzadeh

    2014-09-01

    Full Text Available The Egyptian cotton leafworm, Spodoptera littoralis (Boisduval (Lepidoptera: Noctuidae damages a wide variety of crops in Middle East. Their hosts include cotton, alfalfa, eggplant, tomato, lettuce, bean and some ornamental crops. The intensive use of broad-spectrum insecticides against S. littoralis has led to the development of resistance to many registered pesticides use for its control. The purpose of the present study is biochemical characterization of digestive enzymes of this pest to gain a better understanding of the digestive physiology. The physiology and biochemistry of the insect digestive enzyme had an important role in the study of novel insecticidal strategies. The Egyptian cotton leafworm alimentary canal consists of a short foregut, a long midgut and a short hindgut. Application of pH indicators showed that alimentary canal was alkaline. Our results showed that activities of gut α-amylase were different in three parts of the insect gut. Also shown the greatest activity of α-amylase observed in the midgut followed by hindgut and foregut, respectively. However, there were not significant differences in activity of the enzyme in the midgut and hindgut. The optimal pH α-amylase in foregut, midgut and hindgut were 10.0. Zymogram analysis of different part of gut showed four bands in midgut, hind gut and two bands in foregut. Therefore, in midgut of S. littoralis, four isoenzymes were present. These results explain why more amylase activity was seen in these regions in the spectrophotometric assay.

  3. Chemical composition and α-amylase inhibitory activity of the essential oil from Sabina chinensis cv. Kaizuca leaves.

    Science.gov (United States)

    Gu, Dongyu; Fang, Chen; Yang, Jiao; Li, Minjing; Liu, Hengming; Yang, Yi

    2018-03-01

    Sabina chinensis cv. Kaizuca (SCK) is a variant of S. chinensis L. The essential oil from its leaves exhibited α-amylase inhibitory activity in vitro and the IC 50 value was 187.08 ± 0.56 μg/mL. Nineteen compounds were identified from this essential oil by gas chromatography-mass spectrometry (GC-MS) analysis. The major compounds identified were bornyl acetate (42.6%), elemol (20.5%), β-myrcene (13.7%) and β-linalool (4.0%). In order to study the reason of the α-amylase inhibitory activity of this essential oil, the identified compounds were docked with α-amylase by molecular docking individually. Among these compounds, γ-eudesmol exhibited the lowest binding energy (-6.73 kcal/mol), followed by α-copaen-11-ol (-6.66 kcal/mol), cubedol (-6.39 kcal/mol) and α-acorenol (-6.12 kcal/mol). The results indicated that these compounds were the active ingredients responsible for the α-amylase inhibitory activity of essential oil from SCK.

  4. α-Amylase as a reliable and convenient measure of sympathetic activity: don't start salivating just yet!

    NARCIS (Netherlands)

    Bosch, J.A.; Veerman, E.C.I.; de Geus, E.J.; Proctor, G.B.

    2011-01-01

    Recent years have seen a growing interest in salivary α-amylase (sAA) as a non-invasive marker for sympathetic nervous system (SNS) activity. Saliva offers many advantages as a biomarker fluid and sAA is one of its most plentiful components. sAA is a digestive enzyme that breaks down starch, which

  5. Determination of antioxidant capacity and a-amylase inhibitory activity of the essential oils from citronella grass and lemongrass

    Science.gov (United States)

    The objective of the present study was to determine the antioxidant capacity of and in vitro a-amylase inhibitory activity of the essential oils extracted from citronella grass and lemongrass. The chemical composition of the extracted essential oils was determined by GC-MS. The antioxidant capacity ...

  6. The Effect of Trans-Chalcone on Amylase Activity, Blood Glucose and Lipid Levels in Diabetic and Non Diabetic Rats

    Directory of Open Access Journals (Sweden)

    M Najafian

    2011-04-01

    Full Text Available Introduction & Objective: Alpha amylase is the most important decomposing enzyme in starch. Digestion and absorption of starch in the intestine can be prevented and also the blood sugar levels can be controlled by restrain and control of alpha amylase. The aim of this study was to evaluate the effect of trans-chalcone on amylase activity, blood glucose and lipid levels in diabetic and non diabetic rats. Materials & Methods: This experimental study was conducted in 1388 at Tehran University of Medical Sciences. Sixty rats were randomly divided to ten equal groups: non diabetic control, diabetic control, four non diabetic experiments and four diabetic experiments. Control groups received grape seed oil and experimental groups received 2, 8,16 and 32 mg/kg of body weight in a period of 24 days with a gastric cannula. Blood sugar, every two days, serum insulin levels in days 0,12, and 24 and at the end of the experiment, lipoproteins and alpha amylase activity were measured.The data were analyzed by one way analysis of variance, ANOVA, followed by Turkey,s test with SPSS soft ware . Results: On average Chalcone reduced 25.5% of blood sugar in normal and diabetic rats. IT also decreased the serum insulin level. On average, chalcone decreased 34.9% of alpha amylase activity in normal and diabetic rats. Following disturbances in lipids metabolism caused by diabetes, this drug improved lipoproteins metabolism and reduced water, food and urine volume. Conclusion: This study shows that trans-Chalcone reduces blood sugar and body weight via inhibition of alpha amylas. Moreover, improvement of lipoprotein metabolism may happen via the inhibitory effect of this drug on hydroxyl methyl glutaryl -COA reductase and phosphodiesterase.

  7. Studies on activity, distribution, and zymogram of protease, α-amylase, and lipase in the paddlefish Polyodon spathula.

    Science.gov (United States)

    Ji, H; Sun, H T; Xiong, D M

    2012-06-01

    A series of biochemical determination and electrophoretic observations have been conducted to analyze the activities and characteristics of protease, α-amylase, and lipase of paddlefish Polyodon spathula. The results obtained have been compared with those of bighead carp (Aristichthys nobilis) and hybrid sturgeon (Huso dauricus ♀ × Acipenser schrenki Brandt ♂), in order to increase available knowledge of the physiological characteristics of this sturgeon species and to gain information with regard to its nutrition. Further, a comparative study of enzymatic activity, distribution, and characterization between commercial feed-reared paddlefish (CG) and natural live food-reared (NG) paddlefish was conducted. Results showed that higher proteolytic activity was observed in the pH range 2.5-3.0 and at a pH of 7.0 for paddlefish. Levels of acid protease activity of paddlefish were similar to that of hybrid sturgeon, and significantly higher than that of bighead carp. The inhibition assay of paddlefish showed that the rate of inhibition of tosyl-phenylalanine chloromethyl ketone was approximately 2.6-fold that of tosyl-lysine chloromethyl ketone. There was no significant difference observed for acid protease activity between PG and CG groups, whereas the activity of alkaline protease, α-amylase, and lipase in the PG group were significantly lower than those in the CG group. The substrate sodium dodecyl sulfate polyacrylamide gel electrophoresis analysis further showed that there were certain types of enzymes, especially α-amylase, with similar molecular mass in the paddlefish and hybrid sturgeon. It can be inferred that acid digestion was main mechanism for protein hydrolysis in paddlefish, as reported for other fishes with a stomach. This indicates that the paddlefish requires higher alkaline protease, α-amylase, and lipase activity to digest natural live food.

  8. 2-Aryl benzimidazoles: Synthesis, In vitro α-amylase inhibitory activity, and molecular docking study.

    Science.gov (United States)

    Adegboye, Akande Akinsola; Khan, Khalid Mohammed; Salar, Uzma; Aboaba, Sherifat Adeyinka; Kanwal; Chigurupati, Sridevi; Fatima, Itrat; Taha, Mohammad; Wadood, Abdul; Mohammad, Jahidul Isalm; Khan, Huma; Perveen, Shahnaz

    2018-04-25

    Despite of many diverse biological activities exhibited by benzimidazole scaffold, it is rarely explored for the α-amylase inhibitory activity. For that purpose, 2-aryl benzimidazole derivatives 1-45 were synthesized and screened for in vitro α-amylase inhibitory activity. Structures of all synthetic compounds were deduced by various spectroscopic techniques. All compounds revealed inhibition potential with IC 50 values of 1.48 ± 0.38-2.99 ± 0.14 μM, when compared to the standard acarbose (IC 50  = 1.46 ± 0.26 μM). Limited SAR suggested that the variation in the inhibitory activities of the compounds are the result of different substitutions on aryl ring. In order to rationalize the binding interactions of most active compounds with the active site of α-amylase enzyme, in silico study was conducted. Copyright © 2018 Elsevier Masson SAS. All rights reserved.

  9. Detergent-compatible bacterial amylases.

    Science.gov (United States)

    Niyonzima, Francois N; More, Sunil S

    2014-10-01

    Proteases, lipases, amylases, and cellulases are enzymes used in detergent formulation to improve the detergency. The amylases are specifically supplemented to the detergent to digest starchy stains. Most of the solid and liquid detergents that are currently manufactured contain alkaline enzymes. The advantages of using alkaline enzymes in the detergent formulation are that they aid in removing tough stains and the process is environmentally friendly since they reduce the use of toxic detergent ingredients. Amylases active at low temperature are preferred as the energy consumption gets reduced, and the whole process becomes cost-effective. Most microbial alkaline amylases are used as detergent ingredients. Various reviews report on the production, purification, characterization, and application of amylases in different industry sectors, but there is no specific review on bacterial or fungal alkaline amylases or detergent-compatible amylases. In this mini-review, an overview on the production and property studies of the detergent bacterial amylases is given, and the stability and compatibility of the alkaline bacterial amylases in the presence of the detergents and the detergent components are highlighted.

  10. LaaA, a novel high-active alkalophilic alpha-amylase from deep-sea bacterium Luteimonas abyssi XH031(T).

    Science.gov (United States)

    Song, Qinghao; Wang, Yan; Yin, Chong; Zhang, Xiao-Hua

    2016-08-01

    Alpha-amylase is a kind of broadly used industrial enzymes, most of which have been exploited from terrestrial organism. Comparatively, alpha-amylase from marine environment was largely undeveloped. In this study, a novel alkalophilic alpha-amylase with high activity, Luteimonas abyssi alpha-amylase (LaaA), was cloned from deep-sea bacterium L. abyssi XH031(T) and expressed in Escherichia coli BL21. The gene has a length of 1428bp and encodes 475 amino acids with a 35-residue signal peptide. The specific activity of LaaA reached 8881U/mg at the optimum pH 9.0, which is obvious higher than other reported alpha-amylase. This enzyme can remain active at pH levels ranging from 6.0 to 11.0 and temperatures below 45°C, retaining high activity even at low temperatures (almost 38% residual activity at 10°C). In addition, 1mM Na(+), K(+), and Mn(2+) enhanced the activity of LaaA. To investigate the function of potential active sites, R227G, D229K, E256Q/H, H327V and D328V mutants were generated, and the results suggested that Arg227, Asp229, Glu256 and Asp328 were total conserved and essential for the activity of alpha-amylase LaaA. This study shows that the alpha-amylase LaaA is an alkali-tolerant and high-active amylase with strong potential for use in detergent industry. Copyright © 2016 Elsevier Inc. All rights reserved.

  11. Polysaccharide with antioxidant, α-amylase inhibitory and ACE inhibitory activities from Momordica charantia.

    Science.gov (United States)

    Tan, Hwee-Feng; Gan, Chee-Yuen

    2016-04-01

    Functional polysaccharide was isolated from Momordica charantia, with a yield of 36% (w/w). M. charantia bioactive polysaccharide (MCBP) was an acidic and branched heteropolysaccharide with a molecular weight of 92 kDa. Fourier transform infrared spectroscopic analysis indicated that MCBP was a pectin-like polysaccharide with an esterification degree of 53% and it contains numerous monosaccharides, predominantly glucose, galactose, and galaturonic acid. The results also showed that MCBP exhibited free radical scavenging activity (31.9%), ferric reducing antioxidant power (0.95 mM), α-amylase inhibition (89.1%), and angiotensin-converting enzyme inhibition (94.1%). In the terms of functionality, MCBP showed a lower water-holding capacity but higher in oil-holding capacity, emulsifying activity and foaming capacity compared to citrus pectin. Scanning electron microscopy images demonstrated that MCBP formed gels with a porous structure, and flow analysis showed that the gel solution exhibited pseudoplastic shear-thinning behavior. These findings indicated that MCBP is a promising functional macromolecular carbohydrate for the food and nutraceutical industries. Copyright © 2016 Elsevier B.V. All rights reserved.

  12. Employing in vitro directed molecular evolution for the selection of α-amylase variant inhibitors with activity toward cotton boll weevil enzyme.

    Science.gov (United States)

    da Silva, Maria Cristina Mattar; Del Sarto, Rafael Perseghini; Lucena, Wagner Alexandre; Rigden, Daniel John; Teixeira, Fabíola Rodrigues; Bezerra, Caroline de Andrade; Albuquerque, Erika Valéria Saliba; Grossi-de-Sa, Maria Fatima

    2013-09-20

    Numerous species of insect pests attack cotton plants, out of which the cotton boll weevil (Anthonomus grandis) is the main insect in Brazil and must be controlled to avert large economic losses. Like other insect pests, A. grandis secretes a high level of α-amylases in the midgut lumen, which are required for digestion of carbohydrates. Thus, α-amylase inhibitors (α-AIs) represent a powerful tool to apply in the control of insect pests. Here, we applied DNA shuffling and phage display techniques and obtained a combinatorial library containing 10⁸ α-AI variant forms. From this library, variants were selected exhibiting in vitro affinity for cotton boll weevil α-amylases. Twenty-six variant sequences were cloned into plant expression vectors and expressed in Arabidopsis thaliana. Transformed plant extracts were assayed in vitro to select specific and potent α-amylase inhibitors against boll weevil amylases. While the wild type inhibitors, used to create the shuffled library, did not inhibit the A. grandis α-amylases, three α-AI mutants, named α-AIC3, α-AIA11 and α-AIG4 revealed high inhibitory activities against A. grandis α-amylases in an in vitro assay. In summary, data reported here shown the potential biotechnology of new α-AI variant genes for cotton boll weevil control. Copyright © 2013 The Authors. Published by Elsevier B.V. All rights reserved.

  13. Salivary alpha amylase activity in human beings of different age groups subjected to psychological stress.

    Science.gov (United States)

    Sahu, Gopal K; Upadhyay, Seema; Panna, Shradha M

    2014-10-01

    Salivary alpha-amylase (sAA) has been proposed as a sensitive non-invasive biomarker for stress-induced changes in the body that reflect the activity of the sympathetic nervous system. Though several experiments have been conducted to determine the validity of this salivary component as a reliable stress marker in human subjects, the effect of stress induced changes on sAA level in different age groups is least studied. This article reports the activity of sAA in human subjects of different age groups subjected to psychological stress induced through stressful video clip. Differences in sAA level based on sex of different age groups under stress have also been studied. A total of 112 subjects consisting of both the male and female subjects, divided into two groups on basis of age were viewed a video clip of corneal transplant surgery as stressor. Activity of sAA from saliva samples of the stressed subjects were measured and compared with the activity of the samples collected from the subjects before viewing the clip. The age ranges of subjects were 18-25 and 40-60 years. The sAA level increased significantly in both the groups after viewing the stressful video. The increase was more pronounced in the younger subjects. The level of sAA was comparatively more in males than females in the respective groups. No significant change in sAA activity was observed after viewing the soothed video clip. Significant increase of sAA level in response to psychological stress suggests that it might act as a reliable sympathetic activity biochemical marker in different stages of human beings.

  14. Nutrient Content, Phytonutrient Composition, Alpha Amylase, Alpha Glucosidase Inhibition Activity and Antioxidant Activity of the Stoechospermum Marginatum Collected in Pre Monsoon Season

    Directory of Open Access Journals (Sweden)

    Reka Palanivel

    2017-03-01

    Full Text Available The objective of this study was to investigate the nutrient content, phytonutrient composition, physicochemical properties, alpha amylase and alpha glucosidase inhibition activity and antioxidant activity of the brown algae Stoechospermum marginatum collected from Gulf of Mannar, Tamil Nadu, India in pre monsoon season (June- September, 2015. Six and eight hours of ethanol and aqueous extract of Stoechospermum marginatum were used for phytonutrient screening, alpha amylase, alpha glucosidase inhibition activity and antioxidant activity. From the results of the study it is understood that Stoechospermum marginatum contain a high amount of carbohydrate, protein, crude fiber and phytonutrients like tannin, flavonoid, saponin, alkaloid, terpenoids, steroid and total phenolic content. The physicochemical properties namely Water absorption and Swelling power were very promising. Alpha amylase and alpha glucosidase inhibition activity was recorded to be high in both aqueous and ethanol extracts of eight hour extraction than in extracts taken from six hours extraction. Antioxidant activity was detected using DPPH, FRAP, beta carotene scavenging and H2O2 assay and found to have a high radical scavenging activity. Stoechospermum marginatum possess a valuable amount of total phenolic content and other phytonutrients and physicochemical properties, it may the reason for the potential inhibition of alpha amylase, alpha glucosidase and antioxidant activity. It is concluded from the study that the brown algae may be incorporated into foods to enhance their nutritional and therapeutic value.

  15. Transcription Factor AREB2 Is Involved in Soluble Sugar Accumulation by Activating Sugar Transporter and Amylase Genes.

    Science.gov (United States)

    Ma, Qi-Jun; Sun, Mei-Hong; Lu, Jing; Liu, Ya-Jing; Hu, Da-Gang; Hao, Yu-Jin

    2017-08-01

    Sugars play important roles in plant growth and development, crop yield and quality, as well as responses to abiotic stresses. Abscisic acid (ABA) is a multifunctional hormone. However, the exact mechanism by which ABA regulates sugar accumulation is largely unknown in plants. Here, we tested the expression profile of several sugar transporter and amylase genes in response to ABA treatment. MdSUT2 and MdAREB2 were isolated and genetically transformed into apple ( Malus domestica ) to investigate their roles in ABA-induced sugar accumulation. The MdAREB2 transcription factor was found to bind to the promoters of the sugar transporter and amylase genes and activate their expression. Both MdAREB2 and MdSUT2 transgenic plants produced more soluble sugars than controls. Furthermore, MdAREB2 promoted the accumulation of sucrose and soluble sugars in an MdSUT2 -dependent manner. Our results demonstrate that the ABA-responsive transcription factor MdAREB2 directly activates the expression of amylase and sugar transporter genes to promote soluble sugar accumulation, suggesting a mechanism by which ABA regulates sugar accumulation in plants. © 2017 American Society of Plant Biologists. All Rights Reserved.

  16. Evaluation of the alpha-amylase activity as an indicator of pasteurization efficiency and microbiological quality of liquid whole eggs.

    Science.gov (United States)

    Silva, Guilherme Resende da; Menezes, Liliane Denize Miranda; Lanza, Isabela Pereira; Oliveira, Daniela Duarte de; Silva, Carla Aparecida; Klein, Roger Wilker Tavares; Assis, Débora Cristina Sampaio de; Cançado, Silvana de Vasconcelos

    2017-09-01

    In order to evaluate the efficiency of the pasteurization process in liquid whole eggs, an UV/visible spectrophotometric method was developed and validated for the assessment of alpha-amylase activity. Samples were collected from 30 lots of raw eggs (n = 30) and divided into three groups: one was reserved for analysis of the raw eggs, the second group was pasteurized at 61.1°C for 3.5 minutes (n = 30), and the third group was pasteurized at 64.4°C for 2.5 minutes (n = 30). In addition to assessing alpha-amylase activity, the microbiological quality of the samples was also evaluated by counting total and thermotolerant coliforms, mesophilic aerobic microorganisms, Staphylococcus spp., and Salmonella spp. The validated spectrophotometric method demonstrated linearity, with a coefficient of determination (R2) greater than 0.99, limits of detection (LOD) and quantification (LOQ) of 0.48 mg kg-1 and 1.16 mg kg-1, respectively, and acceptable precision and accuracy with relative standard deviation (RSD) values of less than 10% and recovery rates between 98.81% and 105.40%. The results for alpha-amylase activity in the raw egg samples showed high enzyme activity due to near-complete hydrolysis of the starch, while in the eggs pasteurized at 61.1°C, partial inactivation of the enzyme was observed. In the samples of whole eggs pasteurized at 64.4°C, starch hydrolysis did not occur due to enzyme inactivation. The results of the microbiological analyses showed a decrease (P pasteurized egg samples according to the two binomials under investigation, compared to the raw egg samples, which showed high rates of contamination (P pasteurization, only one sample (3.33%) was positive for Salmonella spp., indicating failure in the pasteurization process, which was confirmed by the alpha-amylase test. It was concluded that the validated methodology for testing alpha-amylase activity is adequate for assessing the efficiency of the pasteurization process, and that the time

  17. Stilbene Glucoside, a Putative Sleep Promoting Constituent from Polygonum multiflorum Affects Sleep Homeostasis by Affecting the Activities of Lactate Dehydrogenase and Salivary Alpha Amylase.

    Science.gov (United States)

    Wei, Qian; Ta, Guang; He, Wenjing; Wang, Wei; Wu, Qiucheng

    2017-01-01

    Chinese herbal medicine (CHM) has been used for treating insomnia for centuries. The most used CHM for insomnia was Polygonum multiflorum. However, the molecular mechanism for CHM preventing insomnia is unknown. Stilbene glucoside (THSG), an important active component of P. multiflorum, may play an important role for treating insomnia. To test the hypothesis, Kunming mice were treated with different dosages of THSG. To examine the sleep duration, a computer-controlled sleep-wake detection system was implemented. Electroencephalogram (EEG) and electromyogram (EMG) electrodes were implanted to determine sleep-wake state. RT-PCR and Western blot was used to measure the levels of lactate dehydrogenase (LDH) and saliva alpha amylase. Spearman's rank correlation coefficient was used to identify the strength of correlation between the variables. The results showed that THSG significantly prolonged the sleep time of the mice (palpha amylase (palpha amylase (pamylase were negatively associated with sleep duration (palpha amylase.

  18. α--AMYLASES OF Aspergillus flavus var. oryzae AND Bacillus subtilis: THE SUBSTRATE SPECIFICITY AND RESISTANCE TO A NUMBER OF CHEMICALLY ACTIVE SUBSTANCES

    Directory of Open Access Journals (Sweden)

    K. V. Avdiyuk

    2013-06-01

    Full Text Available The ability of Aspergillus flavus var. oryzae 80428 and Bacillus subtilis 147 α-amylases to split different carbohydrate-containing substrates, such as maltose, sucrose, trehalose, dextrin, α- and β-cyclodextrin, amylose, amylopectin, glycogen, pullulan, soluble starch, insoluble starch, corn starch, wheat starch, dextran 500 has been studied. It was shown that investigated enzymes differ by substrate specificity. α-Amylase of A. flavus var. oryzae 80428 rapidly hydrolysed soluble potato and wheat starch, while the α-amylase of B. subtilis 147 — only wheat starch. Both enzymes don’t cleave maltose, α-cyclodextrin and dextran 500. A. flavus var. oryzae 80428 α-amylase display very small ability to hydrolyze pullulan, while α-amylase of B. subtilis 147 it does not act in general. The lowest values of Michaelis constant for both enzymes at splitting of glycogen have been obtained, indicating that enzymes have the greatest affinity to this substrate. The studies of influence of chemically active substances on activity of A. flavus var. oryzae 80428 and B. subtilis 147 ?-amylases show there are resistant to urea, deoxycholic acid, Tween-80, Triton X-100 and hydrogen peroxide. It’s indicate the enzymes tested may be competitive in compare with earlier described in literature enzymes. The obtained results give a possibility to propose in future usage these enzymes in different fields of industry, foremost in detergent industry.

  19. Proteinase, amylase, and proteinase-inhibitor activities in the gut of six cockroach species

    Czech Academy of Sciences Publication Activity Database

    Vinokurov, Konstantin; Taranushenko, Yuliya; Krishnan, Natraj; Sehnal, František

    2007-01-01

    Roč. 53, - (2007), s. 794-802 ISSN 0022-1910 R&D Projects: GA ČR(CZ) GA522/06/1591; GA MŠk 1M06030 Institutional research plan: CEZ:AV0Z50070508 Keywords : amylases * Blattodea * gut pH Subject RIV: CE - Biochemistry Impact factor: 2.294, year: 2007

  20. Production of Active Bacillus licheniformis Alpha-Amylase in Tobacco and its Application in Starch Liquefaction

    NARCIS (Netherlands)

    Pen, J; MOLENDIJK, L; Quax, Wim J.; SIJMONS, PC; VANOOYEN, AJJ; VANDENELZEN, PJM; RIETVELD, K; HOEKEMA, A

    As a first example of the feasibility of producing industrial bulk enzymes in plants, we have expressed Bacillus licheniformis alpha-amylase in transgenic tobacco, and applied the seeds directly in starch liquification. The enzyme was properly secreted into the intercellular space, and maximum

  1. Adipokinetic hormones control amylase activity in the cockroach (Periplaneta americana) gut

    Czech Academy of Sciences Publication Activity Database

    Bodláková, K.; Jedlička, Pavel; Kodrík, Dalibor

    2017-01-01

    Roč. 24, č. 2 (2017), s. 259-269 ISSN 1672-9609 R&D Projects: GA ČR GA14-07172S Institutional support: RVO:61388963 ; RVO:60077344 Keywords : AKH * AKH receptor * amylase * enzyme * gene expression * midgut Subject RIV: ED - Physiology OBOR OECD: Entomology; Biochemistry and molecular biology (BC-A) Impact factor: 2.026, year: 2016

  2. Effects of a dietary Aspergillus oryzae extract containing alpha-amylase activity on performance and carcass characteristics of finishing beef cattle.

    Science.gov (United States)

    Tricarico, J M; Abney, M D; Galyean, M L; Rivera, J D; Hanson, K C; McLeod, K R; Harmon, D L

    2007-03-01

    Three experiments were conducted to examine the effects of an Aspergillus oryzae extract containing alpha-amylase activity on performance and carcass characteristics of finishing beef cattle. In Exp. 1, 120 crossbred steers were used in a randomized complete block design to evaluate the effects of roughage source (alfalfa hay vs. cottonseed hulls) and supplemental alpha-amylase at 950 dextrinizing units (DU)/kg of DM. Significant roughage source x alpha-amylase interactions (P < 0.05) were observed for performance. In steers fed cottonseed hulls, supplemental alpha-amylase increased ADG through d 28 and 112 and tended (P < 0.15) to increase ADG in all other periods. The increases in ADG were related to increased DMI and efficiency of gain during the initial 28-d period but were primarily related to increased DMI as the feeding period progressed. Supplemental alpha-amylase increased (P = 0.02) the LM area across both roughage sources. In Exp. 2, 96 crossbred heifers were used in a randomized complete block design with a 2 x 3 factorial arrangement of treatments to evaluate the effects of corn processing (dry cracked vs. high moisture) and supplemental alpha-amylase concentration (0, 580, or 1,160 DU/kg of DM). Alpha-amylase supplementation increased DMI (P = 0.05) and ADG (P = 0.03) during the initial 28 d on feed and carcass-adjusted ADG (P = 0.04) across corn processing methods. Longissimus muscle area was greatest (quadratic effect, P = 0.04), and yield grade was least (quadratic effect, P = 0.02) in heifers fed 580 DU of alpha-amylase/kg of DM across corn processing methods. In Exp. 3, 56 crossbred steers were used in a randomized complete block design to evaluate the effects of supplemental alpha-amylase (930 DU/kg of DM) on performance when DMI was restricted to yield a programmed ADG. Alpha-amylase supplementation did not affect performance when DMI was restricted. We conclude that dietary alpha-amylase supplementation of finishing beef diets may result in

  3. Indole-3-acetic acid (IAA) producing Pseudomonas isolates inhibit seed germination and α-amylase activity in durum wheat (Triticum turgidum L.)

    Energy Technology Data Exchange (ETDEWEB)

    Tabatabaei, S.; Ehsanzadeh, P.; Etesami, H.; Alikhani, H.A.; Glick, B.R.

    2016-11-01

    The role of plant-associated bacteria in plant physiology and metabolism is well documented, but little has been known about the roles played by Pseudomonas in durum wheat (Triticum turgidum L. var durum) growth and development. An in vitroexperiment was conducted to observe the effect of the inoculation of four indole-3-acetic acid (IAA)-producing Pseudomonas isolates and exogenous IAA on seed germination traits and α-amylase activity of durum wheat. The results showed inoculation with all bacterial isolates led to a decrease in the germination percent, although the extent of the depression varied with the isolate. A significant relationship between concentrations of bacterial IAA and the germination inhibition percent in durum wheat seeds by different bacteria strains was observed. The results of this assay showed the effect of bacterial isolates on α-amylase activity after six and 8 days of inoculation was significant, while effect of these isolates on α-amylase activity after two and 4 days of inoculation was not meaningful. In addition, the exogenously applied IAA displayed a concentration-dependent effect on seed germination attributes and α-amylase activity, consistent with the possibility that the inhibitory effect of bacterial inoculation on seed germination was in consequence of bacteria-produced IAA. Therefore, it may suggested that the inhibitory role of IAA in seed germination and α-amylase activity should be taken into account during the screening of IAA-producing Pseudomonas isolates for durum wheat growth promoting agents. (Author)

  4. Salivary type alpha-amylase activity in serum and in urine of patients with lung adenocarcinoma; Aktywnosc alfa-amylazy sliniankowej w surowicy i moczu chorych na gruczolakoraka pluca

    Energy Technology Data Exchange (ETDEWEB)

    Zakrzewska, I.; Wolska, K.; Koput, A. [Zaklad Laboratoryjnej Diagnostyki Klinicznej, Akademia Medyczna, Bialystok (Poland)

    1993-12-31

    Total alpha-amylase activity in sera and urine of 30 patients with lung adenocarcinoma has been tested. The results were compared with control group of 30 healthy voluntaries. The activity of pancreatic type was differentiated from salivary alpha amylase. Salivary type was inhibited selectively by Triticum aestivum. Higher levels of total and salivary type amylase were noted in patients with lung adenocarcinoma in comparison to healthy control. The increase was significant (p<0.005). Correlation was observed between the activity of salivary type amylase and the stage of adenocarcinoma. (author). 12 refs, 3 figs, 1 tab.

  5. Gamma irradiation of sorghum flour: Effects on microbial inactivation, amylase activity, fermentability, viscosity and starch granule structure

    International Nuclear Information System (INIS)

    Mukisa, Ivan M.; Muyanja, Charles M.B.K.; Byaruhanga, Yusuf B.; Schüller, Reidar B.; Langsrud, Thor; Narvhus, Judith A.

    2012-01-01

    Malted and un-malted sorghum (Sorghum bicolor (L.) Moench) flour was gamma irradiated with a dose of 10 kGy and then re-irradiated with 25 kGy. The effects of irradiation on microbial decontamination, amylase activity, fermentability (using an amylolytic L. plantarum MNC 21 strain), starch granule structure and viscosity were determined. Standard methods were used during determinations. The 10 kGy dose had no effect on microbial load of un-malted flour but reduced that of malted flour by 3 log cycles. Re-irradiation resulted in complete decontamination. Irradiation of malt caused a significant (p<0.05) reduction in alpha and beta amylase activity (22% and 32%, respectively). Irradiation of un-malted flour increased the rates of utilization of glucose and maltose by 53% and 100%, respectively, during fermentation. However, microbial growth, rate of lactic acid production, final lactic acid concentration and pH were not affected. Starch granules appeared normal externally even after re-irradiation, however, granules ruptured and dissolved easily after hydration and gelatinization. Production of high dry matter density porridge (200 g dry matter/L) with a viscosity of 3500 cP was achieved by irradiation of un-malted flout at 10 kGy. Gamma irradiation can be used to decontaminate flours and could be utilized to produce weaning porridge from sorghum. - Highlights: ► Malted and un-malted Sorghum flours irradiated (10 kGy) and re-irradiated (25 kGy). ► Complete decontamination only achieved after re-irradiation. ► Significant reduction (p<0.05) in malt amylase activity. ► Microbial growth, starch breakdown and acidification unaffected during fermentation. ► Viscosity of sorghum porridge lowered due to weakened starch granules.

  6. Gamma irradiation of sorghum flour: Effects on microbial inactivation, amylase activity, fermentability, viscosity and starch granule structure

    Science.gov (United States)

    Mukisa, Ivan M.; Muyanja, Charles M. B. K.; Byaruhanga, Yusuf B.; Schüller, Reidar B.; Langsrud, Thor; Narvhus, Judith A.

    2012-03-01

    Malted and un-malted sorghum ( Sorghum bicolor (L.) Moench) flour was gamma irradiated with a dose of 10 kGy and then re-irradiated with 25 kGy. The effects of irradiation on microbial decontamination, amylase activity, fermentability (using an amylolytic L. plantarum MNC 21 strain), starch granule structure and viscosity were determined. Standard methods were used during determinations. The 10 kGy dose had no effect on microbial load of un-malted flour but reduced that of malted flour by 3 log cycles. Re-irradiation resulted in complete decontamination. Irradiation of malt caused a significant ( palpha and beta amylase activity (22% and 32%, respectively). Irradiation of un-malted flour increased the rates of utilization of glucose and maltose by 53% and 100%, respectively, during fermentation. However, microbial growth, rate of lactic acid production, final lactic acid concentration and pH were not affected. Starch granules appeared normal externally even after re-irradiation, however, granules ruptured and dissolved easily after hydration and gelatinization. Production of high dry matter density porridge (200 g dry matter/L) with a viscosity of 3500 cP was achieved by irradiation of un-malted flout at 10 kGy. Gamma irradiation can be used to decontaminate flours and could be utilized to produce weaning porridge from sorghum.

  7. ON THE STRUCTURE OF THE MEDIAN SEGMENT OF THE DIGESTIVE TUBE AND ON THE ACTIVITY OF ALPHA-AMYLASE AT ARISTICHTHYS NOBILIS, IN VARIOUS STAGES OF DEVELOPMENT

    Directory of Open Access Journals (Sweden)

    Gabriela Vasile

    2007-12-01

    Full Text Available The investigations have been dedicated to the structure of the median segment of the digestive tube, on Aristichthys nobilis individuals of various ages (i.e., one summer- and, respectively, four summer-old, correlated, on one side, with the main feeding regime and, on the other, with the amylasic activity present in this part of the digestive tract. As to the structural aspects, higher values of the thickness of the mucous tunic have been recorded in the four summer-old bighead carp individuals, to be possibly associated with the high values of the α-amylase activity in this segment of the digestive tube.

  8. Estimated environmental loads of alpha-amylase from transgenic high-amylase maize

    Energy Technology Data Exchange (ETDEWEB)

    Wolt, Jeffrey D. [Department of Agronomy, Iowa State University, Ames, IA 50011 (United States); Biosafety Institute for Genetically Modified Agricultural Products, 164 Seed Science, Iowa State University, Ames, IA 50011 (United States); Karaman, Sule [Biosafety Institute for Genetically Modified Agricultural Products, 164 Seed Science, Iowa State University, Ames, IA 50011 (United States)

    2007-11-15

    Environmental exposure of plants bioengineered to improve efficiencies of biofuel production is an important consideration for their adoption. High-amylase maize genetically engineered to produce thermostable alpha-amylase in seed endosperm is currently in development, and its successful adoption will entail >1000 km{sup 2} of annual production in the USA. Environmental exposure of thermostable amylase will occur in production fields from preharvest and harvest dropped grain, with minor additional contributions from stover and root biomass. Mass loadings of thermostable alpha-amylase are projected to be 16 kg km{sup -2} and represent a potential source of increased alpha-amylase activity in receiving soils. An understanding of the degradation, persistence, accumulation, and activity of thermostable alpha-amylase introduced from transgenic high-amylase maize will be necessary in order to effectively manage transgenic crop systems intended or biofeedstock production. (author)

  9. Halophilic Amylase from a Moderately Halophilic Micrococcus

    Science.gov (United States)

    Onishi, Hiroshi

    1972-01-01

    A moderately halophilic Micrococcus sp., isolated from unrefined solar salt, produced a considerable amount of extracellular dextrinogenic amylase when cultivated aerobically in media containing 1 to 3 m NaCl. The Micrococcus amylase had maximal activity at pH 6 to 7 in 1.4 to 2 m NaCl or KCl at 50 C. Calcium ion and a high concentration of NaCl or KCl were essential for activity and stability of the amylase. The salt response of the amylase depended greatly on the pH and temperature of the enzyme assay. PMID:5058445

  10. α-Amylase as a reliable and convenient measure of sympathetic activity: don't start salivating just yet!

    Science.gov (United States)

    Bosch, Jos A; Veerman, Enno C I; de Geus, Eco J; Proctor, Gordon B

    2011-05-01

    Recent years have seen a growing interest in salivary α-amylase (sAA) as a non-invasive marker for sympathetic nervous system (SNS) activity. Saliva offers many advantages as a biomarker fluid and sAA is one of its most plentiful components. sAA is a digestive enzyme that breaks down starch, which provides a simple means of quantification by measuring its enzymatic activity. This commentary will address a number of common misconceptions and methodological issues that surround the use of sAA as a marker of SNS activity and limit its utility in biobehavioral research. The usefulness of sAA as an SNS marker is undermined by the fact that the parasympathetic nerves also play a significant role in sAA release. Local parasympathetic nerves regulate sAA activity via: (1) α-amylase release from glands that are solely or mainly parasympathetically innervated; (2) via synergistic sympathetic-parasympathetic effects on protein secretion (known as 'augmented secretion'); and (3) via effects on salivary flow rate. Regarding methodology, we discuss why it is problematic: (1) to ignore the contribution of salivary flow rate; (2) to use absorbent materials for saliva collection, and; (3) to stimulate saliva secretion by chewing. While these methodological problems can be addressed by using standardized and timed collection of unstimulated saliva, the physiological regulation of sAA secretion presents less resolvable issues. We conclude that at present there is insufficient support for the use and interpretation of sAA activity as a valid and reliable measure of SNS activity. Copyright © 2011 Elsevier Ltd. All rights reserved.

  11. Glucan, Water Dikinase Activity Stimulates Breakdown of Starch Granules by Plastidial β-Amylases1[W][OA

    Science.gov (United States)

    Edner, Christoph; Li, Jing; Albrecht, Tanja; Mahlow, Sebastian; Hejazi, Mahdi; Hussain, Hasnain; Kaplan, Fatma; Guy, Charles; Smith, Steven M.; Steup, Martin; Ritte, Gerhard

    2007-01-01

    Glucan phosphorylating enzymes are required for normal mobilization of starch in leaves of Arabidopsis (Arabidopsis thaliana) and potato (Solanum tuberosum), but mechanisms underlying this dependency are unknown. Using two different activity assays, we aimed to identify starch degrading enzymes from Arabidopsis, whose activity is affected by glucan phosphorylation. Breakdown of granular starch by a protein fraction purified from leaf extracts increased approximately 2-fold if the granules were simultaneously phosphorylated by recombinant potato glucan, water dikinase (GWD). Using matrix-assisted laser-desorption ionization mass spectrometry several putative starch-related enzymes were identified in this fraction, among them β-AMYLASE1 (BAM1; At3g23920) and ISOAMYLASE3 (ISA3; At4g09020). Experiments using purified recombinant enzymes showed that BAM1 activity with granules similarly increased under conditions of simultaneous starch phosphorylation. Purified recombinant potato ISA3 (StISA3) did not attack the granular starch significantly with or without glucan phosphorylation. However, starch breakdown by a mixture of BAM1 and StISA3 was 2 times higher than that by BAM1 alone and was further enhanced in the presence of GWD and ATP. Similar to BAM1, maltose release from granular starch by purified recombinant BAM3 (At4g17090), another plastid-localized β-amylase isoform, increased 2- to 3-fold if the granules were simultaneously phosphorylated by GWD. BAM activity in turn strongly stimulated the GWD-catalyzed phosphorylation. The interdependence between the activities of GWD and BAMs offers an explanation for the severe starch excess phenotype of GWD-deficient mutants. PMID:17631522

  12. Vicilin-like peptides from Capsicum baccatum L. seeds are α-amylase inhibitors and exhibit antifungal activity against important yeasts in medical mycology.

    Science.gov (United States)

    Vieira Bard, Gabriela C; Nascimento, Viviane V; Oliveira, Antônia Elenir A; Rodrigues, Rosana; Da Cunha, Maura; Dias, Germana B; Vasconcelos, Ilka M; Carvalho, Andre O; Gomes, Valdirene M

    2014-07-01

    The objective of this study was to isolate antimicrobial peptides from Capsicum baccatum seeds and evaluate their antimicrobial activity and inhibitory effects against α-amylase. Initially, proteins from the flour of C. baccatum seeds were extracted in sodium phosphate buffer, pH 5.4, and precipitated with ammonium sulfate at 90% saturation. The D1 and D2 fractions were subjected to antifungal tests against the yeasts Saccharomyces cerevisiae, Candida albicans, Candida tropicalis, and Kluyveromyces marxiannus, and tested against α-amylases from Callosobruchus maculates and human saliva. The D2 fraction presented higher antimicrobial activity and was subjected to further purification and seven new different fractions (H1-H7) were obtained. Peptides in the H4 fraction were sequenced and the N-terminal sequences revealed homology with previously reported storage vicilins from seeds. The H4 fraction exhibited strong antifungal activity and also promoted morphological changes in yeast, including pseudohyphae formation. All fractions, including H4, inhibited mammalian α-amylase activity but only the H4 fraction was able to inhibit C. maculatus α-amylase activity. These results suggest that the fractions isolated from the seeds of C. baccatum can act directly in plant defenses against pathogens and insects. © 2014 Wiley Periodicals, Inc.

  13. Production of α-amylase from some thermophilic Aspergillus species ...

    African Journals Online (AJOL)

    In this study, thermostable amylase activities of some thermophilic Aspergillus species were evaluated. The suitable medium and microorganisms for α-amylase synthesis were selected. Subsequently, the α-amylase activity of the microorganism was researched. In the measurements made on the 7th day of production on ...

  14. In vitro and in vivo α-amylase and α-glucosidase inhibiting activities of the protein extracts from two varieties of bitter gourd (Momordica charantia L.).

    Science.gov (United States)

    Poovitha, Sundar; Parani, Madasamy

    2016-07-18

    α-amylase and α-glucosidase digest the carbohydrates and increase the postprandial glucose level in diabetic patients. Inhibiting the activity of these two enzymes can control postprandial hyperglycemia, and reduce the risk of developing diabetes. Bitter gourd or balsam pear is one of the important medicinal plants used for controlling postprandial hyperglycemia in diabetes patients. However, there is limited information available on the presence of α-amylase and α-glucosidase inhibiting compounds. In the current study, the protein extracts from the fruits of M. charantia var. charantia (MCC) and M. charantia var. muricata (MCM) were tested for α-amylase and α-glucosidase inhibiting activities in vitro, and glucose lowering activity after oral administration in vivo. The protein extract from both MCC and MCM inhibited the activity of α-amylase and α-glucosidase through competitive inhibition, which was on par with Acarbose as indicated by in vitro percentage of inhibition (66 to 69 %) and IC50 (0.26 to 0.29 mg/ml). Both the protein extracts significantly reduced peak blood glucose and area under the curve in Streptozotocin-induced diabetic rats, which were orally challenged with starch and sucrose. Protein extracts from the fruits of the two varieties of bitter gourd inhibited α-amylase and α-glucosidase in vitro and lowered the blood glucose level in vivo on par with Acarbose when orally administrated to Streptozotocin-induced diabetic rats. Further studies on mechanism of action and methods of safe and biologically active delivery will help to develop an anti-diabetic oral protein drug from these plants.

  15. Radioimmunoassay for plasma renin activity

    International Nuclear Information System (INIS)

    1975-01-01

    A radioimmunoassay for the determination of renin activity in blood plasma is described. The plasma sample is mixed with a generator buffer solution also containing an inhibitor for enzymes which convert angiotensin I into other substances. The renin in the plasma sample converts angiotensinogen into angiotensin I. The amount of angiotensin I is then measured with a competitive binding method using 125 I-labelled angiotensin I and antibodies to angiotensin I

  16. Lactucaxanthin - a potential anti-diabetic carotenoid from lettuce (Lactuca sativa) inhibits α-amylase and α-glucosidase activity in vitro and in diabetic rats.

    Science.gov (United States)

    Gopal, Sowmya Shree; Lakshmi, Magisetty Jhansi; Sharavana, Gurunathan; Sathaiah, Gunaseelan; Sreerama, Yadahally N; Baskaran, Vallikannan

    2017-03-22

    Intestinal and pancreatic α-amylase and α-glucosidase inhibitors offer an approach to lower the levels of post-prandial hyperglycemia through the control of dietary starch breakdown in digestion. This study hypothesized that lactucaxanthin (Lxn) in lettuce (Lactuca sativa) inhibits the activity of α-amylase and α-glucosidase. In this study, the interaction of Lxn with α-amylase and α-glucosidase in silico and its inhibitory effect on these enzymes were studied using in vitro and STZ-induced diabetic rat models. Lxn was isolated from lettuce with 96% purity confirmed by HPLC and LCMS. The in silico analysis showed that Lxn has a lower binding energy (-6.05 and -6.34 kcal mol -1 ) with α-amylase and α-glucosidase compared to their synthetic inhibitors, acarbose (-0.21 kcal mol -1 ) and miglitol (-2.78 kcal mol -1 ), respectively. In vitro α-amylase and α-glucosidase inhibition assays revealed that Lxn had IC 50 values of 435.5 μg mL -1 and 1.84 mg mL -1 , but acarbose has values of 2.5 and 16.19 μg mL -1 . The in vivo results showed an increased activity for α-amylase and α-glucosidase in the intestine (4.7 and 1.30 fold, p < 0.05) and pancreas (1.3 and 1.48 fold, p < 0.05) of STZ induced diabetic rats compared to normal rats. Whereas the activity decreased (p < 0.05) in the Lxn fed diabetic rats, except for the intestinal α-glucosidase activity (1.69 ± 0.12 PNP per min per mg protein). This was confirmed by the low blood glucose level (239.4 ± 18.2 mg dL -1 ) in diabetic rats fed Lxn compared to the diabetic group (572.2 ± 30.5 mg dL -1 , p < 0.05). Lxn significantly inhibited (p < 0.05) the activity of α-amylase and α-glucosidase and could be of medical and nutritional relevance in the treatment of diabetes.

  17. Longitudinal and immediate effect of Kundalini Yoga on salivary levels of cortisol and activity of alpha-amylase and its effect on perceived stress

    Directory of Open Access Journals (Sweden)

    Jocelyn N García-Sesnich

    2017-01-01

    Full Text Available Context: Stress is defined as an alteration of an organism's balance in response to a demand perceived from the environment. Diverse methods exist to evaluate physiological response. A noninvasive method is salivary measurement of cortisol and alpha-amylase. A growing body of evidence suggests that the regular practice of Yoga would be an effective treatment for stress. Aims: To determine the Kundalini Yoga (KY effect, immediate and after 3 months of regular practice, on the perception of psychological stress and the salivary levels of cortisol and alpha-amylase activity. Settings and Design: To determine the psychological perceived stress, levels of cortisol and alpha-amylase activity in saliva, and compare between the participants to KY classes performed for 3 months and a group that does not practice any type of yoga. Subjects and Methods: The total sample consisted of 26 people between 18 and 45-year-old; 13 taking part in KY classes given at the Faculty of Dentistry, University of Chile and 13 controls. Salivary samples were collected, enzyme-linked immunosorbent assay was performed to quantify cortisol and kinetic reaction test was made to determine alpha-amylase activity. Perceived Stress Scale was applied at the beginning and at the end of the intervention. Statistical Analysis Used: Statistical analysis was applied using Stata v11.1 software. Shapiro–Wilk test was used to determine data distribution. The paired analysis was fulfilled by t-test or Wilcoxon signed-rank test. T-test or Mann–Whitney's test was applied to compare longitudinal data. A statistical significance was considered when P< 0.05. Results: KY practice had an immediate effect on salivary cortisol. The activity of alpha-amylase did not show significant changes. A significant decrease of perceived stress in the study group was found. Conclusions: KY practice shows an immediate effect on salivary cortisol levels and on perceived stress after 3 months of practice.

  18. Longitudinal and Immediate Effect of Kundalini Yoga on Salivary Levels of Cortisol and Activity of Alpha-Amylase and Its Effect on Perceived Stress.

    Science.gov (United States)

    García-Sesnich, Jocelyn N; Flores, Mauricio Garrido; Ríos, Marcela Hernández; Aravena, Jorge Gamonal

    2017-01-01

    Stress is defined as an alteration of an organism's balance in response to a demand perceived from the environment. Diverse methods exist to evaluate physiological response. A noninvasive method is salivary measurement of cortisol and alpha-amylase. A growing body of evidence suggests that the regular practice of Yoga would be an effective treatment for stress. To determine the Kundalini Yoga (KY) effect, immediate and after 3 months of regular practice, on the perception of psychological stress and the salivary levels of cortisol and alpha-amylase activity. To determine the psychological perceived stress, levels of cortisol and alpha-amylase activity in saliva, and compare between the participants to KY classes performed for 3 months and a group that does not practice any type of yoga. The total sample consisted of 26 people between 18 and 45-year-old; 13 taking part in KY classes given at the Faculty of Dentistry, University of Chile and 13 controls. Salivary samples were collected, enzyme-linked immunosorbent assay was performed to quantify cortisol and kinetic reaction test was made to determine alpha-amylase activity. Perceived Stress Scale was applied at the beginning and at the end of the intervention. Statistical analysis was applied using Stata v11.1 software. Shapiro-Wilk test was used to determine data distribution. The paired analysis was fulfilled by t -test or Wilcoxon signed-rank test. T -test or Mann-Whitney's test was applied to compare longitudinal data. A statistical significance was considered when P < 0.05. KY practice had an immediate effect on salivary cortisol. The activity of alpha-amylase did not show significant changes. A significant decrease of perceived stress in the study group was found. KY practice shows an immediate effect on salivary cortisol levels and on perceived stress after 3 months of practice.

  19. Alternative method for quantification of alfa-amylase activity Alternativa para quantificação de alfa-amilase

    Directory of Open Access Journals (Sweden)

    DF. Farias

    2010-05-01

    Full Text Available A modification of the sensitive agar diffusion method was developed for macro-scale determination of alfa-amylase. The proposed modifications lower costs with the utilisation of starch as substrate and agar as supporting medium. Thus, a standard curve was built using alfa-amylase solution from Aspergillus oryzae, with concentrations ranging from 2.4 to 7,500 U.mL-1. Clear radial diffusion zones were measured after 4 hours of incubation at 20 °C. A linear relationship between the logarithm of enzyme activities and the area of clear zones was obtained. The method was validated by testing α-amylase from barley at the concentrations of 2.4; 60; 300 and 1,500 U.mL-1. The proposed method turned out to be simpler, faster, less expensive and able to determine on a macro-scale α-amylase over a wide range (2.4 to 7,500 U.mL-1 in scientific investigation as well as in teaching laboratory activities.Modificações foram propostas ao método sensível de difusão em ágar para a macrodeterminação de alfa-amilase. As modificações propostas diminuem os custos, com a utilização de amido como substrato e ágar como meio solidificante. Assim, foi construída uma curva padrão utilizando uma solução de alfa-amilase de Aspergillus oryzae com concentrações variando de 2,4 a 7.500 U.mL-1. Em seguida, as zonas claras de difusão radial foram mensuradas depois de 4 horas de incubação a 20 °C. Foi obtida uma relação linear entre o logaritmo da atividade enzimática e os diâmetros das zonas claras. O método foi validado utilizando-se soluções de alfa-amilase de cevada nas concentrações de 2,4; 60; 300 e 1.500 U.mL-1. O método tornou-se mais simples, rápido, com baixo custo e passível de ser utilizado para macrodeterminação de alfa-amilase em ampla faixa (2,4 a 7.500 U.mL-1 na investigação científica e para fins didáticos em aulas práticas.

  20. Akttvitas Selulase, Amilase Dan Invertase Pada Tanah Kebun Biologi Wamena*[cellulase, Amylase and Invertase Activities Achieved From Soil of Wamena Biological Research Station

    OpenAIRE

    Rahmansyah, M; Latupapua, HJD

    2003-01-01

    Enzymatic activities in soil as due to microbes action in organic matter degradation, lead to propose as indicators for determining soil degree enrichment.In this work, the enzymatic activities of cellulase, invertase and amylase were determined in tropical soil collected from Biological Research Station in Wamena. Result of measurement on five soil samples showed that cellulase activity occurred between 0.10 - 0.31 mg reducing sugar/g soil/hour in 2% Carboxymethylcellulose (CMC) substrate, a...

  1. A single gene directs synthesis of a precursor protein with beta- and alpha-amylase activities in Bacillus polymyxa.

    OpenAIRE

    Uozumi, N; Sakurai, K; Sasaki, T; Takekawa, S; Yamagata, H; Tsukagoshi, N; Udaka, S

    1989-01-01

    The Bacillus polymyxa amylase gene comprises 3,588 nucleotides. The mature amylase comprises 1,161 amino acids with a molecular weight of 127,314. The gene appeared to be divided into two portions by the direct-repeat sequence located at almost the middle of the gene. The 5' region upstream of the direct-repeat sequence was shown to be responsible for the synthesis of beta-amylase. The 3' region downstream of the direct-repeat sequence contained four sequences homologous with those in other a...

  2. Ethanol extract of mango (Mangifera indica L.) peel inhibits α-amylase and α-glucosidase activities, and ameliorates diabetes related biochemical parameters in streptozotocin (STZ)-induced diabetic rats.

    Science.gov (United States)

    Gondi, Mahendranath; Prasada Rao, U J S

    2015-12-01

    Peel is a major by-product during processing of mango fruit into pulp. Recent report indicates that the whole peel powder ameliorated diabetes. In the present study, ethanolic extract of mango peel was analysed for its bioactive compounds, evaluated for α-amylase and α-glucosidase inhibitory properties, oral glucose tolerance test, antioxidant properties, plasma insulin level and biochemical parameters related to diabetes. In addition to gallic and protocatechuic acids, the extract also had chlorogenic and ferulic acids, which were not reported earlier in mango peel extracts. The peel extract inhibited α-amylase and α-glucosidase activities, with IC50 values of 4.0 and 3.5 μg/ml. Ethanolic extract of peel showed better glucose utilization in oral glucose tolerance test. Treatment of streptozotocin-induced diabetic rats with the extract decreased fasting blood glucose, fructosamine and glycated hemoglobin levels, and increased plasma insulin level. Peel extract treatment decreased malondialdehyde level, but increased the activities of antioxidant enzymes significantly in liver and kidney compared to diabetic rats. These beneficial effects were comparable to metformin, but better than gallic acid treated diabetic rats. The beneficial effects of peel extract may be through different mechanism like increased plasma insulin levels, decreased oxidative stress and inhibition of carbohydrate hydrolyzing enzyme activities by its bioactive compounds. Thus, results suggest that the peel extract can be a potential source of nutraceutical or can be used in functional foods and this is the first report on antidiabetic properties of mango peel extract.

  3. Physics of Space Plasma Activity

    International Nuclear Information System (INIS)

    Cramer, N F

    2007-01-01

    This book provides a timely review of our present understanding of plasma phenomena in magnetized terrestrial and solar space plasmas. The author's emphasis is on the fluid and particle modeling and interpretation of observed active processes in space plasmas, i.e. 'the physical background of large plasma eruptions in space'. It is somewhat alarming for a plasma physicist to read that an emphasis on processes in spatially inhomogeneous plasmas means that the work '... excludes a considerable fraction of the available methods in space plasma physics, such as the theory of waves, instabilities and wave particle interactions on a homogeneous background', particularly in light of the fact that much of our knowledge of these plasmas is derived from observations of such waves. However, it is clear on reading the book that such a restriction is not a disadvantage, but allows the author to concentrate on the main theme of the book, namely the use of fluid and particle pictures to model the equilibrium and active states of space plasmas. There are many other books which cover the wave aspects of space plasmas, and would complement this book. The book's coverage is based on the extensive and profound research of the author and his colleagues in the area of fluid and particle modeling of space plasma structures. After an introduction to the physical setting of active plasmas, and a necessarily concise, but effective, discussion of the fluid and particle models to be used, the steady states of the magnetized plasmas of interest are treated, including the magnetosphere, solar plasmas and current sheets. Next the dynamics of unstable states is covered, including MHD and tearing instabilities, and nonlinear aspects, with a detailed discussion of magnetic reconnection. Finally, the models are applied to magnetospheric and solar observations. The book is attractively written and produced, and this reviewer managed to find a minimum number of errors. A particularly attractive

  4. Effect of steeping temperature on antioxidant and inhibitory activities of green tea extracts against α-amylase, α-glucosidase and intestinal glucose uptake.

    Science.gov (United States)

    Liu, Shuyuan; Ai, Zeyi; Qu, Fengfeng; Chen, Yuqiong; Ni, Dejiang

    2017-11-01

    The objective of the present study was to evaluate the effect of steeping temperature on the biological activities of green tea, including the 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical-scavenging capacity, α-glucosidase and α-amylase inhibitory activities, and glucose uptake inhibitory activity in Caco-2 cells. Results showed that, with increasing extraction temperature, the polyphenol content increased, which contributed to enhance antioxidant activity and inhibitory effects on α-glucosidase and α-amylase. Green tea steeped at 100°C showed the highest DPPH radical-scavenging activity and inhibitory effects on α-glucosidase and α-amylase activities with EC 50 or IC 50 values of 6.15μg/mL, 0.09mg/mL, and 6.31mg/mL, respectively. However, the inhibitory potential on glucose uptake did not show an upward trend with increasing extraction temperature. Green tea steeped at 60°C had significantly stronger glucose uptake inhibitory activity (ptea. Copyright © 2017 Elsevier Ltd. All rights reserved.

  5. Rational Engineering of a Cold-Adapted α-Amylase from the Antarctic Ciliate Euplotes focardii for Simultaneous Improvement of Thermostability and Catalytic Activity.

    Science.gov (United States)

    Yang, Guang; Yao, Hua; Mozzicafreddo, Matteo; Ballarini, Patrizia; Pucciarelli, Sandra; Miceli, Cristina

    2017-07-01

    The α-amylases are endo-acting enzymes that hydrolyze starch by randomly cleaving the 1,4-α-d-glucosidic linkages between the adjacent glucose units in a linear amylose chain. They have significant advantages in a wide range of applications, particularly in the food industry. The eukaryotic α-amylase isolated from the Antarctic ciliated protozoon Euplotes focardii ( Ef Amy) is an alkaline enzyme, different from most of the α-amylases characterized so far. Furthermore, Ef Amy has the characteristics of a psychrophilic α-amylase, such as the highest hydrolytic activity at a low temperature and high thermolability, which is the major drawback of cold-active enzymes in industrial applications. In this work, we applied site-directed mutagenesis combined with rational design to generate a cold-active Ef Amy with improved thermostability and catalytic efficiency at low temperatures. We engineered two Ef Amy mutants. In one mutant, we introduced Pro residues on the A and B domains in surface loops. In the second mutant, we changed Val residues to Thr close to the catalytic site. The aim of these substitutions was to rigidify the molecular structure of the enzyme. Furthermore, we also analyzed mutants containing these combined substitutions. Biochemical enzymatic assays of engineered versions of Ef Amy revealed that the combination of mutations at the surface loops increased the thermostability and catalytic efficiency of the enzyme. The possible mechanisms responsible for the changes in the biochemical properties are discussed by analyzing the three-dimensional structural model. IMPORTANCE Cold-adapted enzymes have high specific activity at low and moderate temperatures, a property that can be extremely useful in various applications as it implies a reduction in energy consumption during the catalyzed reaction. However, the concurrent high thermolability of cold-adapted enzymes often limits their applications in industrial processes. The α-amylase from the

  6. Characterisation, immunolocalisation and antifungal activity of a lipid transfer protein from chili pepper (Capsicum annuum) seeds with novel α-amylase inhibitory properties.

    Science.gov (United States)

    Diz, Mariângela S; Carvalho, Andre O; Ribeiro, Suzanna F F; Da Cunha, Maura; Beltramini, Leila; Rodrigues, Rosana; Nascimento, Viviane V; Machado, Olga L T; Gomes, Valdirene M

    2011-07-01

    Lipid transfer proteins (LTPs) were thus named because they facilitate the transfer of lipids between membranes in vitro. This study was triggered by the characterization of a 9-kDa LTP from Capsicum annuum seeds that we call Ca-LTP(1) . Ca-LTP(1) was repurified, and in the last chromatographic purification step, propanol was used as the solvent in place of acetonitrile to maintain the protein's biological activity. Bidimensional electrophoresis of the 9-kDa band, which corresponds to the purified Ca-LTP(1) , showed the presence of three isoforms with isoelectric points (pIs) of 6.0, 8.5 and 9.5. Circular dichroism (CD) analysis suggested a predominance of α-helices, as expected for the structure of an LTP family member. LTPs immunorelated to Ca-LTP(1) from C. annuum were also detected by western blotting in exudates released from C. annuum seeds and also in other Capsicum species. The tissue and subcellular localization of Ca-LTP(1) indicated that it was mainly localized within dense vesicles. In addition, isolated Ca-LTP(1) exhibited antifungal activity against Colletotrichum lindemunthianum, and especially against Candida tropicalis, causing several morphological changes to the cells including the formation of pseudohyphae. Ca-LTP(1) also caused the yeast plasma membrane to be permeable to the dye SYTOX green, as verified by fluorescence microscopy. We also found that Ca-LTP(1) is able to inhibit mammalian α-amylase activity in vitro. Copyright © Physiologia Plantarum 2011.

  7. The Enzyme-Like Domain of Arabidopsis Nuclear β-Amylases Is Critical for DNA Sequence Recognition and Transcriptional Activation.

    Science.gov (United States)

    Soyk, Sebastian; Simková, Klára; Zürcher, Evelyne; Luginbühl, Leonie; Brand, Luise H; Vaughan, Cara K; Wanke, Dierk; Zeeman, Samuel C

    2014-04-01

    Plant BZR1-BAM transcription factors contain a β-amylase (BAM)-like domain, characteristic of proteins involved in starch breakdown. The enzyme-derived domains appear to be noncatalytic, but they determine the function of the two Arabidopsis thaliana BZR1-BAM isoforms (BAM7 and BAM8) during transcriptional initiation. Removal or swapping of the BAM domains demonstrates that the BAM7 BAM domain restricts DNA binding and transcriptional activation, while the BAM8 BAM domain allows both activities. Furthermore, we demonstrate that BAM7 and BAM8 interact on the protein level and cooperate during transcriptional regulation. Site-directed mutagenesis of residues in the BAM domain of BAM8 shows that its function as a transcriptional activator is independent of catalysis but requires an intact substrate binding site, suggesting it may bind a ligand. Microarray experiments with plants overexpressing truncated versions lacking the BAM domain indicate that the pseudo-enzymatic domain increases selectivity for the preferred cis-regulatory element BBRE (BZR1-BAM Responsive Element). Side specificity toward the G-box may allow crosstalk to other signaling networks. This work highlights the importance of the enzyme-derived domain of BZR1-BAMs, supporting their potential role as metabolic sensors. © 2014 American Society of Plant Biologists. All rights reserved.

  8. Effects of radiation and α-tocopherol on saliva flow rate, amylase activity, total protein and electrolyte levels in oral cavity cancer

    Directory of Open Access Journals (Sweden)

    Chitra S

    2008-01-01

    Full Text Available Objective: The objective of the present study was to evaluate early and late effects of radiation and a-tocopherol on the secretion rate of saliva and on selected saliva salivary parameters in oral cavity cancer patients. Patients & Methods: Eighty-nine histologically confirmed oral cavity cancer patients (OCC were enrolled in the study. Resting whole saliva was collected before, during and at the end of the radiation therapy (RT and simultaneous supplementation with α - tocopherol to the radiation treated patients (RT + AT. Results: Salivary flow rate, pH, amylase activity, total protein, sodium and potassium were analyzed. Increased pH, potassium and decreased flow rate, amylase activity, protein content and sodium were observed in 6 weeks of radiation treated patients when compared to OCC patients. A significant improvement of those parameters was observed on α - tocopherol supplementation in RT + AT patients. Conclusion: Supplementation with α - tocopherol improves the salivary flow rate thereby, maintains salivary parameters.

  9. A simple microplate-based method for the determination of α-amylase activity using the glucose assay kit (GOD method).

    Science.gov (United States)

    Visvanathan, Rizliya; Jayathilake, Chathuni; Liyanage, Ruvini

    2016-11-15

    For the first time, a reliable, simple, rapid and high-throughput analytical method for the detection and quantification of α-amylase inhibitory activity using the glucose assay kit was developed. The new method facilitates rapid screening of a large number of samples, reduces labor, time and reagents and is also suitable for kinetic studies. This method is based on the reaction of maltose with glucose oxidase (GOD) and the development of a red quinone. The test is done in microtitre plates with a total volume of 260μL and an assay time of 40min including the pre-incubation steps. The new method is tested for linearity, sensitivity, precision, reproducibility and applicability. The new method is also compared with the most commonly used 3,5-dinitrosalicylic acid (DNSA) method for determining α-amylase activity. Copyright © 2016 Elsevier Ltd. All rights reserved.

  10. ALKALINE PROTEASE, AMYLASE AND CELLULASE ACTIVITIES OF YELLOW RASBORA, Rasbora lateristriataBlkr., AT DIFFERENT FEEDING LEVELS

    Directory of Open Access Journals (Sweden)

    Untung Susilo

    2016-11-01

    Full Text Available Alkaline proteases, amylase and cellulase activities of digestive organ of yellow rasbora, Rasbora lateristriata Blkr., was evaluated with four different feeding levels of 0.34g protein+0.03g fiber, 1.01g protein+0.10g fiber, 1.69g protein+0.16g fiber and 2.36g protein+0.23g fiber/day/100 g fish biomass. A total of 280 fish with average body weight of 0.71±0.06g were used in this study. The results showed that the difference in the feeding levels resulted in a significant difference in trypsin and chymotrypsin  activities (P .05. In conclusion, protein digestion capacity increased, but not to the digestion of starch and fiber in response to different feeding levels and the optimal feeding level for yellow rasbora was 1,01g protein+0,10g fiber/day/100 g fish biomass.

  11. Sol-gel immobilization as a suitable technique for enhancement of α-amylase activity of Aspergillus oryzae PP.

    Science.gov (United States)

    Evstatieva, Yana; Yordanova, Mariya; Chernev, Georgi; Ruseva, Yanislava; Nikolova, Dilyana

    2014-07-04

    Bioencapsulation of microbial cells in silica-based matrices has proved to be a good strategy to enhance the biosynthetic capabilities and viability of bioproducers. In the present study, mycelium and pellet cultures of strain Aspergillus oryzae PP were successfully immobilized in sol-gel hybrid matrices composed of tetraethylorthosilicate as an inorganic precursor, 5% (w/v) starch and 10 or 15% (w/v) polyethylene oxide, or 10% (w/v) calcium alginate as organic compounds. Biosynthetic activity of immobilized cultures was investigated by batch and fed-batch cultivation and the obtained results of 3042.04 IU cm -3 were comparable with the enzyme activity of the free cell culture. Immobilized cultures retained their viability and biosynthetic capabilities up to the 744th h during fed-batch fermentation processes. Consequently, sol-gel encapsulation in hybrid matrices could be considered as a promising technique for immobilization of Aspergillus oryzae PP in order to increase the α-amylase production.

  12. Effect of gamma radiation on the activity of alpha and beta amylases in germinating fenugreek (Trigonella foenum-Graceum, L.) beans

    International Nuclear Information System (INIS)

    Yacoub, N.J.

    1978-01-01

    In previous experiments, three weeks old cotyledons from gamma irradiated seeds of fenugreek beans were observed to be healthier and heavier than their non-irradiated controls. A hypothesis was set that this phenomenon was due to the accumulation of carbohydrate in the cotyledons. To test this hypothesis an experiment was designed to determine the sugars and starch contents in the cotyledons of an irradiated and nonirradiated groups at different stages of development. At the same time the amylases activities in the same groups were assayed. Results indicated that there was actually increased amount of sugars ad starch in the two and three week old cotyledons of the irradiated as compared to their controls group. These findings support the above hypothesis. The amylase activities were also higher in the irradiated groups as compared to their non-irradiated controls. These results suggest that irradiation induced stimulation of the photosynthetic rate, and amylases' activities, meanwhile, slowed the translocation of carbohydrate nutrients from the cotyledons into the growing seedlings

  13. Purification and characterization of peptides from Capsicum annuum fruits which are α-amylase inhibitors and exhibit high antimicrobial activity against fungi of agronomic importance.

    Science.gov (United States)

    Dos Santos, Layrana de Azevedo; Taveira, Gabriel Bonan; Ribeiro, Suzanna de Fátima Ferreira; Pereira, Lídia da Silva; Carvalho, André de Oliveira; Rodrigues, Rosana; Oliveira, Antônia Elenir Amâncio; Machado, Olga Lima Tavares; Araújo, Jucélia da Silva; Vasconcelos, Ilka Maria; Gomes, Valdirene Moreira

    2017-04-01

    Proteins extracted from Capsicum annuum L. fruits were initially subjected to reversed-phase chromatography on HPLC, resulting in eight peptide-rich fractions. All the fractions obtained were tested for their ability to inhibit porcine trypsin and amylase from both human saliva and from larval insect in vitro. All fractions were also tested for their ability to inhibit growth of the phytopathogenic fungi. Several fractions inhibited the activity of human salivary amylase and larval insect amylase, especially fraction Fa5. No fraction tested was found to inhibit trypsin activity, being Fa2 fraction an exception. Interestingly fraction Fa5 also displayed high antimicrobial activity against the species of the Fusarium genus. Fraction Fa5 was found to have two major protein bands of 17 and 6.5 kDa, and these were sequenced by mass spectrometry. Two peptides were obtained from the 6.5-kDa band, which showed similarity to antimicrobial peptides. Fraction Fa5 was also tested for its ability to permeabilize membranes and induce ROS. Fraction Fa5 was able to permeabilize the membranes of all the fungi tested. Fungi belonging to the genus Fusarium also showed an increase in the endogenous production of ROS when treated with this fraction. Antimicrobial peptides were also identified in the fruits from other Capsicum species. Copyright © 2017 Elsevier Inc. All rights reserved.

  14. Amylase and protease inhibition activity against Fusarium verticillioides Amylase and protease inhibition activity against Fusarium verticillioides/ Atividade inibidora de amilase e protease de milho contra Fusarium verticillioides durante a germinação

    Directory of Open Access Journals (Sweden)

    Elisa Yoko Hirooka

    2006-06-01

    Full Text Available The primary maize pathogen, Fusarium verticillioides (F. moniliforme Sheldon is responsible for fumonisin production, which is harmful to human and animal health. In addition, maize can be more susceptible to fungal infection after insect attack. The activity of amylase and protease inhibitors in AG 5011 and CD 307 hybrids were determined during germination with controlled and not controlled conditions of temperature and relative humidity and, they were correlated to maize resistance against Sithophilus zeamais. The inhibitory activity during corn germination was evaluated at 0, 24, 48, 72, 96 and 168 h. Amylase and protease inhibitory activity increased during germination in both hybrids, which ranged respectively from 2.8 to 39.5 UIA/g, and 550.0 to 3633.9 UIP/g. The highest levels of inhibitory activity occurred in hybrid CD 307 in germination chamber. The biologic cycle and susceptible rate were evaluated for corn resistance test. The AG 5011 hybrid was less susceptible to S. zeamais and showed higher inhibitory activity (time 0 h, demonstrating possible relationship between resistance against the insect and inhibitory enzymes. These results indicated that maize natural defense mechanism plays an important role on phytopathogen control.Fusarium verticillioides, patógeno primário do milho, destaca-se pela produção da fumonisina, prejudicial à saúde humana e animal. Considerando que os mecanismos naturais de defesa são ferramentas promissoras no controle de fitopatógenos, avaliou-se: a atividade dos inibidores de amilase e protease presente nos híbridos de milho AG 5011 e CD 307 durante a germinação em câmara de germinação (25ºC e 90-95%UR e em casa de vegetação (sem controle de temperatura e umidade contra amilase e protease de F. verticillioides. Paralelamente, avaliou-se a resistência do milho a Sitophilus zeamais. A atividade inibitória de enzimas avaliada nos tempos 0, 24, 48, 72, 96 e 168 h, aumentou durante a germina

  15. Heart rate, salivary α-amylase activity, and cooperative behavior in previously naïve children receiving dental local anesthesia.

    Science.gov (United States)

    Arhakis, Aristidis; Menexes, George; Coolidge, Trilby; Kalfas, Sotirios

    2012-01-01

    Psychosomatic indicators, such as heart rate (HR), salivary alpha amylase (sAA) activity, and behavior, can be used to determine stress. This study's aim was to assess the pattern of changes of salivary alpha amylase, heart rate, and cooperative behavior in previously naïve children receiving dental treatment under local anesthesia. Included were 30 children with no prior dental experience who needed 4 or more sessions of dental treatment involving local anesthesia. In each session, sAA, HR, and behavior were assessed before and during the application of local anesthesia and at the end of the treatment. The highest sAA value was always observed at the end of each session; overall, the value was lower in the fourth session. HR always increased during the local anesthesia, and did not vary across sessions. No significant relationship was found between child cooperation and either sAA or HR. In this sample, child cooperation may not be an accurate indicator of stress. Based on salivary alpha amylase activity changes, dental treatment involving local anesthesia in naïve children appeared to be less stressful after 3 sessions.

  16. [Amylase production by Aureobasidium pullulans in liquid and solid media].

    Science.gov (United States)

    Lodato, P B; Forchiassin, F; Segovia de Huergo, M B

    1997-01-01

    Amylase production by a strain of Aureobasidium pullulans isolated in the laboratory was evaluated in liquid media (complex and synthetic) and in solid medium (wheat bran). There was an inhibitory effect in amylase production or amylase secretion by glucose. Asparagine was the best nitrogen source for amylase production (4-6 g/l). Only chlamidospores and melanin but not, amylase activity, were obtained with ammonium sulfate. Amylase production in solid culture was higher than the production obtained in the liquid media assayed. Optimum initial moisture content in solid culture ranged between 57 and 74%. No difference was observed in amylase production between solid media inoculated with cells grown in liquid or solid media.

  17. Alternative method for quantification of alfa-amylase activity Alternativa para quantificação de alfa-amilase

    OpenAIRE

    DF. Farias; AFU. Carvalho; CC. Oliveira; NM. Sousa; LCB. Rocha-Bezerrra; PMP. Ferreira; GPG. Lima; DC. Hissa

    2010-01-01

    A modification of the sensitive agar diffusion method was developed for macro-scale determination of alfa-amylase. The proposed modifications lower costs with the utilisation of starch as substrate and agar as supporting medium. Thus, a standard curve was built using alfa-amylase solution from Aspergillus oryzae, with concentrations ranging from 2.4 to 7,500 U.mL-1. Clear radial diffusion zones were measured after 4 hours of incubation at 20 °C. A linear relationship between the logarithm of...

  18. Influência do fumo na atividade da amilase salivar e na curva glicêmica Influence of smoking on salivary amylase activity and glycemic curve

    Directory of Open Access Journals (Sweden)

    Patricia Moriel

    2010-08-01

    Full Text Available OBJETIVO: Determinar a atividade da amilase salivar e a relação com a glicemia, antes e após a ingestão de carboidratos em fumantes e não fumantes, uma vez que in vitro a exposição da saliva à fumaça do cigarro induz à redução da atividade da amilase salivar e poderia influenciar na absorção dos carboidratos da dieta. MÉTODOS: Foram avaliados voluntários fumantes (n=10 e não fumantes (n=10. Realizou-se coletas da saliva antes e após o fumo e determinou-se a glicemia antes e após a ingestão de 72g de carboidratos. Para glicemia usaram-se tempos de 0, 15, 30, 60, 120 minutos. A determinação da atividade da amilase salivar foi realizada por meio de kits comerciais. A glicemia foi determinada utilizando o aparelho Glicomiter (Accu-Chek-Roche. As análises estatísticas foram realizadas no software Sigmastat, utilizou-se o método Teste t pareado (pOBJECTIVE: The objective of this study was to determine salivary amylase activity and its relationship with glycemia before and after smokers and nonsmokers ingested carbohydrates. Since cigarette smoke reduces salivary amylase activity in vitro, it may affect dietary carbohydrate absorption. METHODS: Twenty volunteers participated in this study, 10 smokers and 10 nonsmokers. Samples of saliva were collected before and after the smokers had a cigarette and glycemia was determined before and after the ingestion of 72g of carbohydrates. Glycemia was measured 0, 15, 30, 60 and 120 minutes after carbohydrate intake. Salivary amylase activity was determined by commercial kits. Glycemia was determined by a glucometer (Accu-Chek-Roche. The paired t-test was used for the statistical analyses, done by the software Sigmastat, with p<0.05. RESULTS: Glycemia 15, 30, 60 and 90 minutes after carbohydrate intake rose 3.9%, 11.9%, 34.8% and 22.7% in nonsmokers and 4.9%, 6.5%, 13.8% and 9.7% in smokers, respectively. The peak glucose absorption in nonsmokers was 21.0% greater than in smokers. Salivary

  19. Production of amylases by Aspergillus tamarii

    Directory of Open Access Journals (Sweden)

    Moreira Fabiana Guillen

    1999-01-01

    Full Text Available A strain of Aspergillus tamarii, a filamentous fungus isolated from soil, was able to produce both a-amylase and glucoamylase activities in mineral media supplemented with 1% (w/v starch or maltose as the carbon source. Static cultivation led to significantly higher yields than those obtained using shaking culture. The production of amylases was tolerant to a wide range of initial culture pH values (from 4 to 10 and temperature (from 25 to 42oC. Two amylases, one a-amylase and one glucoamylase, were separated by ion exchange chromatography. Both partially purified enzymes had optimal activities at pH values between 4.5 and 6.0 and were stable under acid conditions (pH 4.0-7.0. The enzymes exhibited optimal activities at temperatures between 50o and 60o C and were stable for more than ten hours at 55oC.

  20. Malaysian brown seaweeds Sargassum siliquosum and Sargassum polycystum: Low density lipoprotein (LDL) oxidation, angiotensin converting enzyme (ACE), α-amylase, and α-glucosidase inhibition activities.

    Science.gov (United States)

    Nagappan, Hemlatha; Pee, Poh Ping; Kee, Sandra Hui Yin; Ow, Ji Tsong; Yan, See Wan; Chew, Lye Yee; Kong, Kin Weng

    2017-09-01

    Two Malaysian brown seaweeds, Sargassum siliquosum and Sargassum polycystum were first extracted using methanol to get the crude extract (CE) and further fractionated to obtain fucoxanthin-rich fraction (FRF). Samples were evaluated for their phenolic, flavonoid, and fucoxanthin contents, as well as their inhibitory activities towards low density lipoprotein (LDL) oxidation, angiotensin converting enzyme (ACE), α-amylase, and α-glucosidase. In LDL oxidation assay, an increasing trend in antioxidant activity was observed as the concentration of FRF (0.04-0.2mg/mL) and CE (0.2-1.0mg/mL) increased, though not statistically significant. As for serum oxidation assay, significant decrease in antioxidant activity was observed as concentration of FRF increased, while CE showed no significant difference in inhibitory activity across the concentrations used. The IC 50 values for ACE inhibitory activity of CE (0.03-0.42mg/mL) were lower than that of FRF (0.94-1.53mg/mL). When compared to reference drug Voglibose (IC 50 value of 0.61mg/mL) in the effectiveness in inhibiting α-amylase, CE (0.58mg/mL) gave significantly lower IC 50 values while FRF (0.68-0.71mg/mL) had significantly higher IC 50 values. The α-glucosidase inhibitory activity of CE (IC 50 value of 0.57-0.69mg/mL) and FRF (IC 50 value of 0.50-0.53mg/mL) were comparable to that of reference drug (IC 50 value of 0.54mg/mL). Results had shown the potential of S. siliquosum and S. polycystum in reducing cardiovascular diseases related risk factors following their inhibitory activities on ACE, α-amylase and α-glucosidase. In addition, it is likelihood that FRF possessed antioxidant activity at low concentration level. Copyright © 2017 Elsevier Ltd. All rights reserved.

  1. Effects of germination on the activities of amylases and phenolic enzymes in sorghum varieties grouped according to food end-use properties

    NARCIS (Netherlands)

    Dicko, M.H.; Gruppen, H.; Zouzouho, O.C.; Traore, A.S.; Berkel, van W.J.H.; Voragen, A.G.J.

    2006-01-01

    Fifty sorghum varieties were screened to determine the effects of germination on levels of starch, -amylase, -amylase, phenylalanine ammonia lyase (PAL), peroxidase (POX) and polyphenol oxidase (PPO). Germination decreased starch content, with amylose being more degraded than amylopectin. In

  2. Lower immunglobulin A levels but not lower cortisol or α-amylase activity in children with chronic tension-type headache.

    Science.gov (United States)

    Fernández-de-Las-Peñas, César; Fernández-Mayoralas, Daniel M; Arroyo-Morales, Manuel; Ambite-Quesada, Silvia; Rivas-Martínez, Inés; Ortega-Santiago, Ricardo; Díaz-Rodríguez, Lourdes; Pareja, Juan A

    2011-03-01

    The study was designed to investigate the differences in salivary cortisol (hypothalamic-pituitary-adrenocortical [HPA] axis), immunoglobulin A (IgA) (immune system) concentrations and α-amylase (sympathetic nervous system [SNS]) activity between children with chronic tension-type headache (CTTH) and healthy children. Thirty-six children, 10 boys and 26 girls (age: 9 ± 2 years) with CTTH and 36 age- and sex-matched healthy children were recruited. Salivary cortisol, α-amylase activity, salivary flow rate, IgA concentration and IgA rate were collected from non-stimulated saliva. A headache diary was used for collecting data on intensity, frequency and duration of headache for four weeks. Children with CTTH showed lower IgA concentration (p = .008) and IgA rate (p = .039), but not lower cortisol concentration (p = .447), salivary flow rate (p = .289) or α-amylase activity (p = .559), as compared to healthy children. Neither age (p > .582) nor gender (p > .227) influenced salivary markers. A significant association between the number of years with headache and IgA concentration (r(s) = - 0.385; p = .023) was found: the greater the number of years with headache, the lower the IgA concentration. These results suggest that children with CTTH present with deficits in the immune system, but not dysfunction in the HPA axis or SNS. Future studies are needed to elucidate the direction of these relationships.

  3. amylase from cockroach, Periplaneta americana

    African Journals Online (AJOL)

    STORAGESEVER

    2008-08-04

    Aug 4, 2008 ... The enzyme was stable up to 55°C and its pH stability was in range of 5.6 - 6.6. .... acrylamide gel and by gel filtration chromatography. Gel filtration .... Activity at 55 °C. Figure 8. Thermic inactivation at 37°C and 55°C of Periplaneta americana α-amylase. Thermal stability of the enzyme was followed for 1 h.

  4. Enzymatic activity and immunoreactivity of Aca s 4, an alpha-amylase allergen from the storage mite Acarus siro

    Czech Academy of Sciences Publication Activity Database

    Pytelková, Jana; Lepšík, Martin; Šanda, Miloslav; Talacko, Pavel; Marešová, Lucie; Mareš, Michael

    2012-01-01

    Roč. 13, č. 3 (2012), s. 1-8 ISSN 1471-2091 R&D Projects: GA ČR GP525/09/P600 Institutional research plan: CEZ:AV0Z40550506 Keywords : Aca s 4 * Acarus siro * alpha - amylase s Subject RIV: CE - Biochemistry Impact factor: 1.776, year: 2012 http://www.biomedcentral.com/1471-2091/13/3

  5. Study of Serum Amylase and Serum Cholinesterase in Organophosphorus Poisoning

    Directory of Open Access Journals (Sweden)

    Sharan Badiger

    2016-04-01

    Full Text Available Background: Poisoning due to organophosphorus compounds is most commonly seen. Earlier plasma cholinesterase level was used to assess the severity of poisoning. Presently serum amylase is being recommended as a better indicator of severity. Aims and Objectives: To study plasma cholinesterase and serum amylase levels in acute organophosphorus and to correlate serum amylase levels with clinical severity and outcome. Material and Methods: A total of 80 patients in the study admitted to a tertiary care centre within 24 hours with a history of organophosphorus poisoning were included in study. Estimation of plasma cholinesterase and serum rd amylase was done at the time of admission, and on 3 th day and on 5 day. Results: Occurrence of organophosphorus poisoning was more common among age group 21-30 years and among males (57.5%. They were 25 (31.2% farmers, 23 (28.8% st u d e n ts, a n d 2 2 ( 2 7 . 5% h o u s ewi v e s. Monocrotophos (45.0% was commonly used compound. Mean value of plasma cholinesterase and serum amylase at admission are 3693 U/L, and 185.4 U/L. There was significant inhibition of plasma cholinesterase and elevation of serum amylase at th admission with return to normal values on 5 day. Conclusion: Plasma cholinesterase inhibition 200 U/L has been associated with poor prognosis and proneness to respiratory failure.

  6. Purification and medium optimization of α-amylase from Bacillus ...

    African Journals Online (AJOL)

    α-Amylase was first time isolated and purified from Bacillus subtilis 168 (1A1). Purified α-amylase fraction showed a single protein band with a molecular weight of 55 kD. Chemical characterization of the purified α-amylase revealed optimum amylolytic activity at 37°C and pH 7.0 using starch as substrate. It was stable at pH ...

  7. Distribution of phenolic antioxidants in whole and milled fractions of quinoa and their inhibitory effects on α-amylase and α-glucosidase activities.

    Science.gov (United States)

    Hemalatha, P; Bomzan, Dikki Pedenla; Sathyendra Rao, B V; Sreerama, Yadahally N

    2016-05-15

    Whole grain quinoa and its milled fractions were evaluated for their phenolic composition in relation to their antioxidant properties and inhibitory effects on α-amylase and α-glucosidase activities. Compositional analysis by HPLC-DAD showed that the distribution of phenolic compounds in quinoa is not entirely localised in the outer layers of the kernel. Milling of whole grain quinoa resulted in about 30% loss of total phenolic content in milled grain. Ferulic and vanillic acids were the principal phenolic acids and rutin and quercetin were predominant flavonoids detected in whole grain and milled fractions. Quinoa milled fractions exhibited numerous antioxidant activities. Despite having relatively lower phenolic contents, dehulled and milled grain fractions showed significantly (p ⩽ 0.05) higher metal chelating activity than other fractions. Furthermore, extracts of bran and hull fractions displayed strong inhibition towards α-amylase [IC50, 108.68 μg/ml (bran) and 148.23 μg/ml (hulls)] and α-glucosidase [IC50, 62.1 μg/ml (bran) and 68.14 μg/ml (hulls)] activities. Thus, whole grain quinoa and its milled fractions may serve as functional food ingredients in gluten-free foods for promoting health. Copyright © 2015 Elsevier Ltd. All rights reserved.

  8. Synthesis, spectroscopic characterization of palladium(II)-orthohydroxyacetophenone azine nano-optical sensor doped in sol–gel matrix and its use as probe for assessment of α-amylase activity in human saliva

    Energy Technology Data Exchange (ETDEWEB)

    El-Sayed, B.A. [Chemistry department, Faculty of Science, al Azhr University, Cairo (Egypt); Abo-Aly, M.M., E-mail: aboalymoh@hotmail.com [Chemistry department, Faculty of Science, Ain Shams University, Cairo (Egypt); Attia, M.S.; Gamal, S. [Chemistry department, Faculty of Science, Ain Shams University, Cairo (Egypt)

    2016-01-15

    PdAPA (palladium(II)-orthohydroxyacetophenone azine) complex doped in sol–gel matrix is synthesized. It is characterized using UV-visible, infrared spectra and TEM image. A novel, simple, sensitive spectrofluorometric method was developed for measuring the activity of the α-amylase enzyme in human saliva for smokers and non-smokers with age range (17–64 years) based on the quenching of the luminescence intensity at 450 nm of the new synthesized complex characterized by various concentrations of the maltose released from the hydrolysis of starch by α-amylase enzyme and was successfully used as nano-optical sensor. The calibration plot was achieved over the concentration range 4.7×10{sup −6}–9.3×10{sup −10} mol L{sup −1} maltose with a correlation coefficient of 0.996 and a detection minimum limit value of 7.55×10{sup −10} mol L{sup −1}. The method was satisfactorily used for the assessment of the α-amylase activity in a number of human saliva samples for various smokers and non-smoker's volunteers. - Highlights: • The (palladium(II)-orthohydroxyacetophenone azine) complex doped in sol–gel was prepared. • It was used for the assessment of of α-amylase enzyme activity. • By maltose resulting from the reaction of α-amylase enzyme with starch. • A novel, simple, sensitive and precise spectrofluorometric method was developed.

  9. Synthesis, spectroscopic characterization of palladium(II)-orthohydroxyacetophenone azine nano-optical sensor doped in sol–gel matrix and its use as probe for assessment of α-amylase activity in human saliva

    International Nuclear Information System (INIS)

    El-Sayed, B.A.; Abo-Aly, M.M.; Attia, M.S.; Gamal, S.

    2016-01-01

    PdAPA (palladium(II)-orthohydroxyacetophenone azine) complex doped in sol–gel matrix is synthesized. It is characterized using UV-visible, infrared spectra and TEM image. A novel, simple, sensitive spectrofluorometric method was developed for measuring the activity of the α-amylase enzyme in human saliva for smokers and non-smokers with age range (17–64 years) based on the quenching of the luminescence intensity at 450 nm of the new synthesized complex characterized by various concentrations of the maltose released from the hydrolysis of starch by α-amylase enzyme and was successfully used as nano-optical sensor. The calibration plot was achieved over the concentration range 4.7×10 −6 –9.3×10 −10 mol L −1 maltose with a correlation coefficient of 0.996 and a detection minimum limit value of 7.55×10 −10 mol L −1 . The method was satisfactorily used for the assessment of the α-amylase activity in a number of human saliva samples for various smokers and non-smoker's volunteers. - Highlights: • The (palladium(II)-orthohydroxyacetophenone azine) complex doped in sol–gel was prepared. • It was used for the assessment of of α-amylase enzyme activity. • By maltose resulting from the reaction of α-amylase enzyme with starch. • A novel, simple, sensitive and precise spectrofluorometric method was developed.

  10. Influences of AMY1 gene copy number and protein expression on salivary alpha-amylase activity before and after citric acid stimulation in splenic asthenia children.

    Science.gov (United States)

    Yang, Zemin; Lin, Jing; Chen, Longhui; Zhang, Min; Yang, Xiaorong; Chen, Weiwen

    2015-06-01

    To compare the correlations between salivary alpha-amylase (sAA) activity and amylase, alpha 1 (salivary) gene (AMYl) copy number or its gene expression between splenic asthenia and healthy children, and investigate the reasons of attenuated sAA activity ratio before and after citric acid stimulation in splenic asthenia children. Saliva samples from 20 splenic asthenia children and 29 healthy children were collected before and after citric acid stimulation. AMYl copy number, sAA activity, and total sAA and glycosylated sAA contents were determined, and their correlations were analyzed. Although splenic asthenia and healthy children had no differences in AMY1 copy number, splenic asthenia children had positive correlations between AMY1 copy number and sAA activity before or after citric acid stimulation. Splenic asthenia children had a higher sAA glycosylated proportion ratio and glycosylated sAA content ratio, while their total sAA content ratio and sAA activity ratio were lower compared with healthy children. The glycosylated sAA content ratio was higher than the total sAA content ratio in both groups. Splenic asthenia and healthy children had positive correlations between total sAA or glycosylated sAA content and sAA activity. However, the role played by glycosylated sAA content in sAA activity in healthy children increased after citric acid stimulation, while it decreased in splenic asthenia children. Genetic factors like AMY1 copy number variations, and more importantly, sAA glycosylation abnormalities leading to attenuated sAA activity after citric acid stimulation, which were the main reasons of the attenuated sAA activity ratio in splenic asthenia children compared with healthy children.

  11. Characterization of a thermostable Bacillus subtilis β-amylase

    African Journals Online (AJOL)

    ... 70 0C respectively, and the thermal stability curve gave a maximum activity of 9.75 U at 70oC for 60 min of incubation. Bacillus subtilis â-amylase is valuable for maltose production, which can be hydrolyzed further by other groups of amylase for the production of high cassava glucose syrup used as sweeteners in the food ...

  12. Nitrogen supplements effect on amylase production by Aspergillus ...

    African Journals Online (AJOL)

    The production of amylase by Aspergillus niger on three cassava whey media in liquid shake culture was compared. The supplemented cassava whey (SCW) medium exhibited gave amylase activity of 495 U/ml. Biomass cropped was 1.63 g/l in the SCW medium. Yeast extract employed as a nitrogen supplement increased ...

  13. Hypoglycemic effect of basil (Ocimum basilicum) aqueous extract is mediated through inhibition of α-glucosidase and α-amylase activities: an in vitro study.

    Science.gov (United States)

    El-Beshbishy, Ha; Bahashwan, Sa

    2012-02-01

    The present study investigated the in vitro hypoglycemic activity of basil (Ocimum basilicum) aqueous extract. Preliminary phytochemical screening of the extract revealed the presence of reducing sugars, cardiac glycosides, tannins, saponins, glycosides, flavonoids and steroids. The total polyphenols content (TPC), flavonoids content (FC), percentage diphenylpicrylhydrazyl (DPPH( · )) radical inhibition and total antioxidant status (TAS) were estimated. The FC was 41 ± 2.2 rutin/g dry extract, the TPC was 146 ± 5.26 mg catechin/g dry extract and the TAS was 5.12 ± 0.7 mmol/L. The %DPPH( · ) free radical inhibition was 60%, 54%, 49% and 43%, respectively, for different extract concentrations; 20, 18.2, 16.3 and 14.5 mg/ml, respectively. The extract elicited significant dose-dependent pattern against rat intestinal sucrase (RIS; IC(50) = 36.72 mg/ml), rat intestinal maltase (RIM; IC(50) = 21.31 mg/ml) and porcine pancreatic α-amylase (PPA; IC(50) = 42.50 mg/ml) inhibitory activities. The inhibition was greater against maltase compared with sucrase. These effects may be attributed to the high TPC and FC levels. The linear regression analysis revealed strong significant positive correlations between %DPPH( · ) radical inhibition and each of %RIS, %RIM and %PPA inhibiting activity. Also, strong significant positive correlations between %RIS and either %RIM or %PPA inhibition activity were observed. We concluded therefore that basil aqueous extract via antioxidant and possibly α-glucosidase and α-amylase inhibiting activities, offered positive benefits to control diabetes.

  14. The role of α-amylase in the perception of oral texture and flavour in custards

    NARCIS (Netherlands)

    Wijk, R.A.de; Prinz, J.F.; Engelen, L.; Weenen, H.

    2004-01-01

    The role of salivary α-amylase in odour, flavour, and oral texture sensations was investigated in two studies in which the activity of salivary amylase present in the mouth of human subjects was either increased by presenting custards with added α-amylase or decreased by presenting custards with

  15. The role of alpha-amylase in the perception of oral texture and flavour in custards

    NARCIS (Netherlands)

    Wijk, de R.A.; Prinz, J.F.; Engelen, L.; Weenen, H.

    2004-01-01

    The role of salivary a-amylase in odour, flavour, and oral texture sensations was investigated in two studies in which the activity of salivary amylase present in the mouth of human subjects was either increased by presenting custards with added alpha-amylase or decreased by presenting custards with

  16. Evaluation of commercial a-amylase enzyme-linked immunosorbent assy (ELISA) test kits for wheat

    Science.gov (United States)

    a-Amylase enzyme is associated with preharvest sprouting (PHS) and late-maturity a amylase (LMA) in wheat, and reduces wheat and flour quality. Various means have been developed to measure the presence of a-amylase, thereby predicting end-use quality; most are based on enzyme activity. An alternativ...

  17. Improvement of heterologous protein production in Aspergillus oryzae by RNA interference with alpha-amylase genes.

    Science.gov (United States)

    Nemoto, Takashi; Maruyama, Jun-ichi; Kitamoto, Katsuhiko

    2009-11-01

    Aspergillus oryzae RIB40 has three alpha-amylase genes (amyA, amyB, and amyC), and secretes alpha-amylase abundantly. However, large amounts of endogenous secretory proteins such as alpha-amylase can compete with heterologous protein in the secretory pathway and decrease its production yields. In this study, we examined the effects of suppression of alpha-amylase on heterologous protein production in A. oryzae, using the bovine chymosin (CHY) as a reporter heterologous protein. The three alpha-amylase genes in A. oryzae have nearly identical DNA sequences from those promoters to the coding regions. Hence we performed silencing of alpha-amylase genes by RNA interference (RNAi) in the A. oryzae CHY producing strain. The silenced strains exhibited a reduction in alpha-amylase activity and an increase in CHY production in the culture medium. This result suggests that suppression of alpha-amylase is effective in heterologous protein production in A. oryzae.

  18. Protease activity of plasma hemopexin

    NARCIS (Netherlands)

    Bakker, WW; Borghuis, T; Harmsen, MC; van den Berg, Anke; Kema, IP; Niezen, KE; Kapojos, JJ

    Background. Previous studies into the relevance of a putative circulating factor in the pathogenesis of minimal change nephrotic syndrome have opened the possibility that plasma hemopexin might be an important effector molecule in this disorder. Thus, intra renal infusion of isolated plasma

  19. Caracterização da atividade amilásica do malte de milho (Zea mays L. = Characterization of amylase activity from maize (Zea mays L. malt

    Directory of Open Access Journals (Sweden)

    Joana Paula Menezes Biazus

    2006-01-01

    Full Text Available Este trabalho objetivou estudar o processo de germinação e a caracterização da atividade bioquímica das amilases do malte de milho (Zea mays L. para gerar uma fonte de amilase de baixo custo e boa atividade enzimática. A atividade enzimática foi monitorada todos os dias durante a germinação das sementes para se obter a melhor condição de produção do malte. Os resultados mostraram que a atividade enzimática nas sementes foi maior no 4º. dia de germinação. A caracterização bioquímica mostrou que as amilases do malte apresentam faixa ótima de pH entre 4,3 e 6, com temperaturas ótimas a 50°C e 80ºC e os valores de Km e Vmax para hidrólise do amido foram de 7,69.10-2 g/L e 7,69.102g/L.min, respectivamente.This work aimed to study the germination process and characterization of the amylolytic activity of the maize (Zea mays L. malt aiming to obtain source amylases at lower cost. Enzymatic activity was monitored all days during the seed germination, for obtaining thebest condition of malt production. Results showed that the enzymatic activity from maize seeds was larger in 4° germination day. Enzymes characterization showed that the maize malt amylases have optimal zone of pH between 4.3 and 6, with optimal temperatures of 50°C and 80ºC. The Km and Vmax values for starch hydrolysis were 7.69.10-2 g/L and 7.69.102g/L.min, respectively.

  20. Influence of Aging on Plasma Renin Activity

    International Nuclear Information System (INIS)

    Cho, K. W.; Kim, S. H.; Kang, S. K.; Choi, H. Y.

    1982-01-01

    Influence of aging on plasma renin activity was evaluated in healthy normotensive subjects(age range 21-63 years, 413 males) devoid of cardiorenal or endocrinological problems. The age-related decrease of plasma renin activity in the subjects between 21-28 years group and 36-42 years group was slight, but over the 43 years groups was significantly different. The age-related suppression of plasma renin activity was much more smooth and continuous all over the age ranges evaluated. The sexual difference in plasma renin activity was noticed between the subjects of 22 years old group (34 males) and 19 years group (34 females) (p<0.003). The data suggest that the age-related suppression of plasma renin activity appeared in healthy normotensive subjects should be considered in the case of evaluation of low renin essential hypertension.

  1. Plasma diamine oxidase activity in asthmatic children

    Directory of Open Access Journals (Sweden)

    Kyoichiro Toyoshima

    1996-01-01

    Full Text Available Histamine plays an important role in the development of asthmatic symptoms. Diamine oxidase (DAO histaminase, which inactivates histamine, is located in the intestine and kidney and is released into plasma. Plasma DAO activity in asthmatic children was measured by a recently developed high performance liquid chromatographic method using histamine as the DAO substrate. Diamine oxidase activity was higher in severely asthmatic children than in those with mild asthma. A time course study during the acute exacerbation phase revealed that DAO activity rose during acute asthmatic attacks and then decreased gradually over several days. Although the mechanisms of plasma DAO activity increase during acute asthmatic attacks could not be explained, data showed that plasma DAO activity is an important index of histamine metabolism in asthmatics and may relate to some mechanisms of acute exacerbation of airway inflammation. Consequently, fluctuations in plasma DAO can be used as one of various indices of instability in management of asthma.

  2. In vitro anti-diabetic activity of flavonoids and pheophytins from Allophylus cominia Sw . on PTP1B, DPPIV, alpha-glucosidase and alpha-amylase enzymes.

    Science.gov (United States)

    Semaan, D G; Igoli, J O; Young, L; Marrero, E; Gray, A I; Rowan, E G

    2017-05-05

    Ethno-botanical information from diabetic patients in Cuba led to the identification of Allophylus cominia as a possible source of new drugs for the treatment of type 2 diabetes mellitus (T2-DM). Chemical characterization of the extracts from A. cominia was carried out using chromatographic and spectroscopic methods. The extracts were tested for their activity on PTP1B, DPPIV, α-glucosidase enzymes and α-amylase. The flavonoid rich fractions from A. cominia inhibited DPPIV enzyme (75.3±2.33%) at 30µg/ml and produced a concentration-dependent inhibition against DPPIV with a Ki value of 2.6µg/ml. At 30µg/ml, flavonoids and pheophytins extracts significantly inhibited PTP1B enzyme (100±2.6% and 68±1% respectively). The flavonoids, pheophytin A and pheophytin B fractions showed significant concentration-dependent inhibition against PTP1B with Ki values of 3µg/ml, 0.64µg/ml and 0.88µg/ml respectively. At 30µg/ml, the flavonoid fraction significantly inhibited α-glucosidase enzyme (86±0.3%) in a concentration-dependent pattern with a Ki value of 2µg/ml. None of the fractions showed significant effects on α-amylase. Fatty acids, tannins, pheophytins A and B, and a mixture of flavonoids were detected in the methanolic extract from A. cominia. The identified flavonoids were mearnsitrin, quercitrin, quercetin-3-alloside, and naringenin-7-glucoside. The pharmacological effects of the extracts from A. cominia earlier observed in experimental diabetic models was confirmed in this study. Thus a new drug or formulation for the treatment of T2-DM could be developed from A. cominia. Copyright © 2017 Elsevier Ireland Ltd. All rights reserved.

  3. Inhibitory effects of rosmarinic acid extracts on porcine pancreatic amylase in vitro.

    Science.gov (United States)

    McCue, Patrick P; Shetty, Kalidas

    2004-01-01

    Porcine pancreatic alpha-amylase (PPA) was allowed to react with herbal extracts containing rosmarinic acid (RA) and purified RA. The derivatized enzyme-phytochemical mixtures obtained were characterized for residual amylase activity. These in vitro experiments showed that the amylase activity was inhibited in the presence of these phytochemicals. The extent of amylase inhibition correlated with increased concentration of RA. RA-containing oregano extracts yielded higher than expected amylase inhibition than similar amount of purified RA, suggesting that other phenolic compounds or phenolic synergies may contribute to additional amylase inhibitory activity. The significance of food-grade, plant-based amylase inhibitors for modulation of diabetes mellitus and other oxidation-linked diseases is hypothesized and discussed.

  4. Contact activation of blood-plasma coagulation

    Science.gov (United States)

    Golas, Avantika

    Surface engineering of biomaterials with improved hemocompatibility is an imperative, given the widespread global need for cardiovascular devices. Research summarized in this dissertation focuses on contact activation of FXII in buffer and blood plasma frequently referred to as autoactivation. The extant theory of contact activation imparts FXII autoactivation ability to negatively charged, hydrophilic surfaces. According to this theory, contact activation of plasma involves assembly of proteins comprising an "activation complex" on activating surfaces mediated by specific chemical interactions between complex proteins and the surface. This work has made key discoveries that significantly improve our core understanding of contact activation and unravel the existing paradigm of plasma coagulation. It is shown herein that contact activation of blood factor XII (FXII, Hageman factor) in neat-buffer solution exhibits a parabolic profile when scaled as a function of silanized-glass-particle activator surface energy (measured as advancing water adhesion tension t°a=g° Iv costheta in dyne/cm, where g°Iv is water interfacial tension in dyne/cm and theta is the advancing contact angle). Nearly equal activation is observed at the extremes of activator water-wetting properties --36 moderated by adsorption of plasma proteins unrelated to coagulation through an "adsorption-dilution" effect that blocks FXII contact with hydrophobic activator surfaces. The adsorption-dilution effect explains the apparent specificity for hydrophilic activators pursued by earlier investigators. Finally a comparison of FXII autoactivation in buffer, serum, protein cocktail, and plasma solutions is shown herein. Activation of blood plasma coagulation in vitro by contact with material surfaces is demonstrably dependent on plasma-volume-to-activator-surface-area ratio. However, activation of factor XII dissolved in buffer, protein cocktail, heat-denatured serum, and FXI deficient plasma does not

  5. Blood amylase - a biochemical radiation indicator?

    International Nuclear Information System (INIS)

    Hofmann, R.; Hendriks, W.; Schreiber, G.A.; Boegl, K.W.

    1990-12-01

    This study describes the suitability of the biological radiation indicator 'amylase in human blood serum' to identify, if previous irradiation has occurred. After 'in vivo' exposure, of the human body with organ doses > 0,5 Gy, high activities of the enzyme are found in serum. The important results of examinations from different work groups and from own experiments were summarized in tabular form and evaluated from the statistic point of view. The results show that in more than 90% of all cases, the amylase system is suitable to identify exposure beyond 0,5 to 1 Gy, approximately. However, this is only possible if the salivary glands were also exposed and blood samples are taken about 18 hours after exposure. For the differentiation between induced increase of amylase from disease and radiation induced increase, it is recommended to carry out the isoamylase test, which makes it possible to distinguish between the individual enzymes. The assessment of the total amylase is appropriate to detect, with a range of significance of P = 0,05 that radiation exposure has occurred. The increase of activity is dose dependent when the salivary glands lie in the radiation field, however, the variations of activity are very high. Therefore the radiation dose cannot be considered. Only in cases where a very high radiation induced increase of activity is observed, a rough estimation of dose at the parotid glands can be made. (orig./MG) [de

  6. Oligosaccharide binding to barley alpha-amylase 1

    DEFF Research Database (Denmark)

    Robert, X.; Haser, R.; Mori, H.

    2005-01-01

    Enzymatic subsite mapping earlier predicted 10 binding subsites in the active site substrate binding cleft of barley alpha-amylase isozymes. The three-dimensional structures of the oligosaccharide complexes with barley alpha-amylase isozyme 1 (AMY1) described here give for the first time a thorough...... in barley alpha-amylase isozyme 2 (AMY2), and the sugar binding modes are compared between the two isozymes. The "sugar tongs" surface binding site discovered in the AMY1-thio-DP4 complex is confirmed in the present work. A site that putatively serves as an entrance for the substrate to the active site...

  7. Thermal stability of α-amylase in aqueous cosolvent systems

    Indian Academy of Sciences (India)

    Prakash

    The activity and thermal stability of α-amylase were studied in the presence of different concentrations of ... 2.1 Materials ..... unfavourable free energy of transfer of amino acid side ..... folded protein with a hydrophobic dye: evidence that molten.

  8. Molecular cloning and biochemical characterization of an α-amylase family from Aspergillus niger

    Directory of Open Access Journals (Sweden)

    Junying Wang

    2018-03-01

    Full Text Available Background: α-Amylase is widely used in the starch processing, food and paper industries, hydrolyzing starch, glycogen and other polysaccharides into glucose, maltose and oligosaccharides. An α-amylase gene family from Aspergillus niger CBS513.88 encode eight putative α-amylases. The differences and similarities, biochemical properties and functional diversity among these eight α-amylases remain unknown. Results: The eight genes were cloned and expressed in Pichia pastoris GS115 by shaking-flask fermentation under the induction of methanol. The sequence alignment, biochemical characterizations and product analysis of starch hydrolysis by these α-amylases were investigated. It is found that the eight α-amylases belonged to three different groups with the typical structure of fungal α-amylase. They exhibited maximal activities at 30–40°C except AmyG and were all stable at acidic pH. Ca2+ and EDTA had no effects on the activities of α-amylases except AmyF and AmyH, indicating that the six amylases were Ca2+ independent. Two novel α-amylases of AmyE and AmyF were found. AmyE hydrolyzed starch into maltose, maltotriose and a small amount of glucose, while AmyF hydrolyzed starch into mainly glucose. The excellent physical and chemical properties including high acidic stability, Ca2+-independent and high maltotriose-forming capacity make AmyE suitable in food and sugar syrup industries. Conclusions: This study illustrates that a gene family can encode multiple enzymes members having remarkable differences in biochemical properties. It provides not only new insights into evolution and functional divergence among different members of an α-amylase family, but the development of new enzymes for industrial application. Keywords: Biochemical properties, Food industry, Fungal α-amylase, Glycosyl hydrolase family, Glycosyl hydrolase family, Industrial application, Paper industry, Recombinant Pichia pastoris, Starch processing, α-amylase cloning

  9. Characterization of a new cell-bound alpha-amylase in Bacillus subtilis 168 Marburg that is only immunologically related to the exocellular alpha-amylase.

    OpenAIRE

    Haddaoui, E; Petit-Glatron, M F; Chambert, R

    1995-01-01

    Immunoblot analysis of Bacillus subtilis cell extracts with polyclonal antibodies, raised against purified exocellular alpha-amylase, revealed one protein species of 82,000 Da. This protein was found even in cells in which the amyE gene, encoding exocellular alpha-amylase, was disrupted. Isolated from the membrane fraction, the 82,000-M(r) protein displayed an alpha-amylase activity in vitro.

  10. Effect of exogenous xylanase, amylase, and protease as single or combined activities on nutrient digestibility and growth performance of broilers fed corn/soy diets.

    Science.gov (United States)

    Amerah, A M; Romero, L F; Awati, A; Ravindran, V

    2017-04-01

    Two trials (a 42-d performance and a 21-d cohort digestibility) were conducted to evaluate the performance and nutrient digestibility of broilers fed corn diets supplemented with exogenous xylanase, amylase, and protease as single or combined activities. A nutritionally adequate, positive control (PC) diet was formulated. The negative control (NC) diet was formulated to be lower in metabolizable energy (∼86 kcal/kg diet) and digestible amino acids (1 to 2%) compared to PC. The other 4 treatments were based on the NC and they were either supplemented with xylanase (X), amylase (A), protease (P), or a combination of X, A, and P (XAP; to provide 2,000 U of X, 200 U of A, and 4,000 U of P/kg diet). All diets were marginal in AvP and Ca and contained a background of phytase (1,000 FTU/kg). In each trial, male broiler (Ross 308) chicks were allocated to the 5 treatments (10 replicates of 20 birds/pen and 9 replicates of 8 birds/cage for the performance and digestibility trials, respectively). In the digestibility trial, ileal digesta was collected on d21 for the determination of nutrient utilization. Data were subjected to one-way ANOVA and means were separated by Tukey's HSD test. Only the XAP improved (P digestibility and apparent ileal digestible energy (AIDE). Both P and XAP improved N retention. The relative improvement in energy digestibility due to enzyme supplementation was greater at the ileal level than that measured in the excreta. The measured changes on AIDE due to supplemental enzymes were much higher than the sum of calculated contributions from starch, fat, and protein. Supplementation of all enzymes reduced (P  0.05) by dietary treatments. Both X and XAP had lower (P  0.05) FCR compared to PC. In conclusion, these results suggest a synergistic effect between X, A and P on broiler performance and nutrient digestibility. In the current study, AIDE measurements appeared to overestimate the enzyme response. Calculation of the energy contribution by

  11. The activity of barley alpha-amylase on starch granules is enhanced by fusion of a starch binding domain from Aspergillus niger glucoamylase

    DEFF Research Database (Denmark)

    Juge, N.; Nøhr, J.; Le Gal-Coëffet, M.-F.

    2006-01-01

    High affinity for starch granules of certain amylolytic enzymes is mediated by a separate starch binding domain (SBD). In Aspergillus niger glucoamylase (GA-I), a 70 amino acid O-glycosylated peptide linker connects SBD with the catalytic domain. A gene was constructed to encode barley alpha......-amylase 1 (AMY1) fused C-terminally to this SBD via a 37 residue GA-I linker segment. AMY1-SBD was expressed in A. niger, secreted using the AMY1 signal sequence at 25 mg x L(-1) and purified in 50% yield. AMY1-SBD contained 23% carbohydrate and consisted of correctly N-terminally processed multiple forms...... in A. niger). AMY1-SBD showed a 2-fold increased activity for soluble starch at low (0.5%) but not at high (1%) concentration. AMY1-SBD hydrolysed amylose DP440 with an increased degree of multiple attack of 3 compared to 1.9 for rAMY1. Remarkably, at low concentration (2 nM), AMY1-SBD hydrolysed...

  12. Amylase Production from Thermophilic Bacillus sp. BCC 021-50 Isolated from a Marine Environment

    Directory of Open Access Journals (Sweden)

    Altaf Ahmed Simair

    2017-06-01

    Full Text Available The high cost of fermentation media is one of the technical barriers in amylase production from microbial sources. Amylase is used in several industrial processes or industries, for example, in the food industry, the saccharification of starchy materials, and in the detergent and textile industry. In this study, marine microorganisms were isolated to identify unique amylase-producing microbes in starch agar medium. More than 50 bacterial strains with positive amylase activity, isolated from marine water and soil, were screened for amylase production in starch agar medium. Bacillus sp. BCC 021-50 was found to be the best amylase-producing strain in starch agar medium and under submerged fermentation conditions. Next, fermentation conditions were optimized for bacterial growth and enzyme production. The highest amylase concentration of 5211 U/mL was obtained after 36 h of incubation at 50 °C, pH 8.0, using 20 g/L molasses as an energy source and 10 g/L peptone as a nitrogen source. From an application perspective, crude amylase was characterized in terms of temperature and pH. Maximum amylase activity was noted at 70 °C and pH 7.50. However, our results show clear advantages for enzyme stability in alkaline pH, high-temperature, and stability in the presence of surfactant, oxidizing, and bleaching agents. This research contributes towards the development of an economical amylase production process using agro-industrial residues.

  13. Stabilization of α-amylase by using anionic surfactant during the immobilization process

    International Nuclear Information System (INIS)

    El-Batal, A.I.; Atia, K.S.; Eid, M.

    2005-01-01

    This work describes the entrapment of α-amylase into butylacrylate-acrylic acid copolymer (BuA/AAc) using γ irradiation. The effect of an anionic surfactant (AOT), the reuse efficiency, and kinetic behavior of immobilized α-amylase were studied. Covering of α-amylase with bis-(2-ethylhexyl)sulfosuccinate sodium salt (AOT) made the enzyme more stable than the uncovered form. The hydrolytic activity of the pre-coated immobilized α-amylase was increased below the critical micelle concentration (cmc) (10mmol/L). The results showed an increase in the relative activity with increase in the degree of hydration. The pre-coated immobilized α-amylase showed a higher k cat /K m and lower activation energy compared to the free and uncoated-immobilized preparation, respectively. The results suggest that the immobilization of α-amylase is a potentially useful approach for commercial starch hydrolysis in two-phase systems

  14. Stabilization of {alpha}-amylase by using anionic surfactant during the immobilization process

    Energy Technology Data Exchange (ETDEWEB)

    El-Batal, A.I. [National Center for Radiation Research and Technology, P.O. Box 29, Nasr City, Cairo (Egypt); Atia, K.S. [Nuclear Research Center, Radioisotopes Applications Division, Abo-Zable, P.O. Box 13759, Cairo (Egypt)]. E-mail: ks_atia@yahoo.com; Eid, M. [National Center for Radiation Research and Technology, P.O. Box 29, Nasr City, Cairo (Egypt)

    2005-10-01

    This work describes the entrapment of {alpha}-amylase into butylacrylate-acrylic acid copolymer (BuA/AAc) using {gamma} irradiation. The effect of an anionic surfactant (AOT), the reuse efficiency, and kinetic behavior of immobilized {alpha}-amylase were studied. Covering of {alpha}-amylase with bis-(2-ethylhexyl)sulfosuccinate sodium salt (AOT) made the enzyme more stable than the uncovered form. The hydrolytic activity of the pre-coated immobilized {alpha}-amylase was increased below the critical micelle concentration (cmc) (10mmol/L). The results showed an increase in the relative activity with increase in the degree of hydration. The pre-coated immobilized {alpha}-amylase showed a higher k{sub cat}/K{sub m} and lower activation energy compared to the free and uncoated-immobilized preparation, respectively. The results suggest that the immobilization of {alpha}-amylase is a potentially useful approach for commercial starch hydrolysis in two-phase systems.

  15. Stabilization of α-amylase by using anionic surfactant during the immobilization process

    Science.gov (United States)

    El-Batal, A. I.; Atia, K. S.; Eid, M.

    2005-10-01

    This work describes the entrapment of α-amylase into butylacrylate-acrylic acid copolymer (BuA/AAc) using γ irradiation. The effect of an anionic surfactant (AOT), the reuse efficiency, and kinetic behavior of immobilized α-amylase were studied. Covering of α-amylase with bis-(2-ethylhexyl)sulfosuccinate sodium salt (AOT) made the enzyme more stable than the uncovered form. The hydrolytic activity of the pre-coated immobilized α-amylase was increased below the critical micelle concentration (cmc) (10 mmol/L). The results showed an increase in the relative activity with increase in the degree of hydration. The pre-coated immobilized α-amylase showed a higher k/K and lower activation energy compared to the free and uncoated-immobilized preparation, respectively. The results suggest that the immobilization of α-amylase is a potentially useful approach for commercial starch hydrolysis in two-phase systems.

  16. Enhanced stability and catalytic activity of immobilized α-amylase on modified Fe3O4 nanoparticles for potential application in food industries

    Science.gov (United States)

    Hosseinipour, Seyyedeh Leila; Khiabani, Mahmoud Sowti; Hamishehkar, Hamed; Salehi, Roya

    2015-09-01

    Enzymes play an essential role in catalyzing various reactions. However, their instability upon repetitive/prolonged use, elevated temperature, acidic or alkaline pH remains an area of concern. α-Amylase, a widely used enzyme in food industries for starch hydrolysis, was covalently immobilized on the surface of two developed matrices, amino-functionalized silica-coated magnetite nanoparticles (AFSMNPs) alone and covered with chitosan. The synthesis steps and characterizations of NPs were examined by FT-IR, VSM, and SEM. Modified nanoparticles with average diameters of 20-80 nm were obtained. Enzyme immobilization efficiencies of 89 and 74 were obtained for AFSMNPs and chitosan-coated AFSMNPs, respectively. The optimum pH obtained was 6.5 and 8.0 for the enzyme immobilized on AFSMNPs and chitosan-coated AFSMNPs, respectively. Optimum temperature for the immobilized enzyme shifted toward higher temperatures. Considerable enhancements in thermal stabilities were observed for the immobilized enzyme at elevated temperatures up to 80 °C. A frequent use experiment demonstrated that the immobilized enzyme retained 74 and 85 % of its original activity even after 20 times of repeated use in AFSMNPs and chitosan-coated AFSMNPs, respectively. Storage stability demonstrated that free enzyme lost its activity completely within 30 days. But, immobilized enzyme on AFSMNPs and chitosan-coated AFSMNPs preserved 65.73 and 78.63 % of its initial activity, respectively, after 80 days of incubation. In conclusion, a substantial improvement in the performance of the immobilized enzyme with reference to the free enzyme was obtained. Furthermore, the relative activities of immobilized enzyme are superior than free enzyme over the broader pH and temperature ranges.

  17. Enhanced stability and catalytic activity of immobilized α-amylase on modified Fe3O4 nanoparticles for potential application in food industries

    International Nuclear Information System (INIS)

    Hosseinipour, Seyyedeh Leila; Khiabani, Mahmoud Sowti; Hamishehkar, Hamed; Salehi, Roya

    2015-01-01

    Enzymes play an essential role in catalyzing various reactions. However, their instability upon repetitive/prolonged use, elevated temperature, acidic or alkaline pH remains an area of concern. α-Amylase, a widely used enzyme in food industries for starch hydrolysis, was covalently immobilized on the surface of two developed matrices, amino-functionalized silica-coated magnetite nanoparticles (AFSMNPs) alone and covered with chitosan. The synthesis steps and characterizations of NPs were examined by FT-IR, VSM, and SEM. Modified nanoparticles with average diameters of 20–80 nm were obtained. Enzyme immobilization efficiencies of 89 and 74 were obtained for AFSMNPs and chitosan-coated AFSMNPs, respectively. The optimum pH obtained was 6.5 and 8.0 for the enzyme immobilized on AFSMNPs and chitosan-coated AFSMNPs, respectively. Optimum temperature for the immobilized enzyme shifted toward higher temperatures. Considerable enhancements in thermal stabilities were observed for the immobilized enzyme at elevated temperatures up to 80 °C. A frequent use experiment demonstrated that the immobilized enzyme retained 74 and 85 % of its original activity even after 20 times of repeated use in AFSMNPs and chitosan-coated AFSMNPs, respectively. Storage stability demonstrated that free enzyme lost its activity completely within 30 days. But, immobilized enzyme on AFSMNPs and chitosan-coated AFSMNPs preserved 65.73 and 78.63 % of its initial activity, respectively, after 80 days of incubation. In conclusion, a substantial improvement in the performance of the immobilized enzyme with reference to the free enzyme was obtained. Furthermore, the relative activities of immobilized enzyme are superior than free enzyme over the broader pH and temperature ranges.

  18. Enhanced stability and catalytic activity of immobilized α-amylase on modified Fe{sub 3}O{sub 4} nanoparticles for potential application in food industries

    Energy Technology Data Exchange (ETDEWEB)

    Hosseinipour, Seyyedeh Leila [Tabriz University of Medical Sciences, Biotechnology Research Center (Iran, Islamic Republic of); Khiabani, Mahmoud Sowti [Tabriz University, Department of Food Science and Technology, Faculty of Agricultural Science (Iran, Islamic Republic of); Hamishehkar, Hamed, E-mail: hamishehkar.hamed@gmail.com [Tabriz University of Medical Sciences, Drug Applied Research Center (Iran, Islamic Republic of); Salehi, Roya, E-mail: salehiro@tbzmed.ac.ir [Tabriz University of Medical Sciences, Research Center for Pharmaceutical Nanotechnology and School of Advanced Medical Science (Iran, Islamic Republic of)

    2015-09-15

    Enzymes play an essential role in catalyzing various reactions. However, their instability upon repetitive/prolonged use, elevated temperature, acidic or alkaline pH remains an area of concern. α-Amylase, a widely used enzyme in food industries for starch hydrolysis, was covalently immobilized on the surface of two developed matrices, amino-functionalized silica-coated magnetite nanoparticles (AFSMNPs) alone and covered with chitosan. The synthesis steps and characterizations of NPs were examined by FT-IR, VSM, and SEM. Modified nanoparticles with average diameters of 20–80 nm were obtained. Enzyme immobilization efficiencies of 89 and 74 were obtained for AFSMNPs and chitosan-coated AFSMNPs, respectively. The optimum pH obtained was 6.5 and 8.0 for the enzyme immobilized on AFSMNPs and chitosan-coated AFSMNPs, respectively. Optimum temperature for the immobilized enzyme shifted toward higher temperatures. Considerable enhancements in thermal stabilities were observed for the immobilized enzyme at elevated temperatures up to 80 °C. A frequent use experiment demonstrated that the immobilized enzyme retained 74 and 85 % of its original activity even after 20 times of repeated use in AFSMNPs and chitosan-coated AFSMNPs, respectively. Storage stability demonstrated that free enzyme lost its activity completely within 30 days. But, immobilized enzyme on AFSMNPs and chitosan-coated AFSMNPs preserved 65.73 and 78.63 % of its initial activity, respectively, after 80 days of incubation. In conclusion, a substantial improvement in the performance of the immobilized enzyme with reference to the free enzyme was obtained. Furthermore, the relative activities of immobilized enzyme are superior than free enzyme over the broader pH and temperature ranges.

  19. α-Amylase and α-glucosidase inhibitory effects of Sclerocarya birrea ...

    African Journals Online (AJOL)

    ajl yemi

    2011-10-26

    Oct 26, 2011 ... birrea stem bark (SBSB) extracts against human urinary α-amylase and Bacillus steatothermophilus α ... levels, reduced plasma cholesterol, triglyceride and urea .... cycle and controlled conditions of temperature and humidity.

  20. Production of Alpha Amylase by Bacillus cereus in Submerged Fermentation

    Directory of Open Access Journals (Sweden)

    Helen H. Raplong

    2014-09-01

    Full Text Available Microorganisms have the ability to secrete enzymes when they are grown in the presence of certain substrates. Amylases are among the most important industrial enzymes and are of great significance in biotechnological studies. Bacteria belonging to the genus Bacillus were isolated using mannitol egg yolk polymyxin B (MYP agar a highly selective media for Bacillus cereus isolation. The isolates were tested for α-amylase production on nutrient agar supplemented with starch and in submerged fermentation. The bacteria isolated and identified (using the Microgen Bacillus identification kit were all Bacillus cereus and SB2 had the largest zone of hydrolysis of 12mm on nutrient agar supplemented with starch as well as the highest enzyme activity of 1.62U/ml. Amylase activity of 2.56U/ml was obtained after 24 hours incubation in submerged fermentation. When amylase enzyme production parameters where optimized, maximum amylase activity was obtained at a pH of 6.5, temperature of 350C, incubation time of 24 hours and 4% inoculums concentration. Bacillus cereus SB2 is a potential isolate for alpha-amylase production with soluble starch as the sole carbon source in submerged fermentation.

  1. An Amylase-Responsive Bolaform Supra-Amphiphile.

    Science.gov (United States)

    Kang, Yuetong; Cai, Zhengguo; Tang, Xiaoyan; Liu, Kai; Wang, Guangtong; Zhang, Xi

    2016-02-01

    An amylase-responsive bolaform supra-amphiphile was constructed by the complexation between β-cyclodextrin and a bolaform covalent amphiphile on the basis of host-guest interaction. The bolaform covalent amphiphile could self-assemble in solution, forming sheet-like aggregates and displaying weak fluorescence because of aggregation-induced quenching. The addition of β-cyclodextrin led to the formation of the bolaform supra-amphiphile, prohibiting the aggregation of the bolaform covalent amphiphile and accompanying with the significant recovery of fluorescence. Upon the addition of α-amylase, with the degradation β-cyclodextrin, the fluorescence of the supra-amphiphile would quench gradually and significantly, and the quenching rate linearly correlated to the concentration of α-amylase. This study enriches the field of supra-amphiphiles on the basis of noncovalent interactions, and moreover, it may provide a facile way to estimate the activity of α-amylase.

  2. Occupation of low-affinity cholecystokinin (CCK) receptors by CCK activates signal transduction and stimulates amylase secretion in pancreatic acinar cells.

    Science.gov (United States)

    Vinayek, R; Patto, R J; Menozzi, D; Gregory, J; Mrozinski, J E; Jensen, R T; Gardner, J D

    1993-03-10

    Based on the effects of monensin on binding of 125I-CCK-8 and its lack of effect on CCK-8-stimulated amylase secretion we previously proposed that pancreatic acinar cells possess three classes of CCK receptors: high-affinity receptors, low-affinity receptors and very low-affinity receptors [1]. In the present study we treated pancreatic acini with carbachol to induce a complete loss of high-affinity CCK receptors and then examined the action of CCK-8 on inositol trisphosphate IP3(1,4,5), cytosolic calcium and amylase secretion in an effort to confirm and extend our previous hypothesis. We found that first incubating pancreatic acini with 10 mM carbachol decreased binding of 125I-CCK-8 measured during a second incubation by causing a complete loss of high-affinity CCK receptors with no change in the low-affinity CCK receptors. Carbachol treatment of acini, however, did not alter the action of CCK-8 on IP3(1,4,5), cytosolic calcium or amylase secretion or the action of CCK-JMV-180 on amylase secretion or on the supramaximal inhibition of amylase secretion caused by CCK-8. The present findings support our previous hypothesis that pancreatic acinar cells possess three classes of CCK receptors and suggest that high-affinity CCK receptors do not mediate the action of CCK-8 on enzyme secretion, that low-affinity CCK receptors may mediate the action of CCK on cytosolic calcium that does not involve IP3(1,4,5) and produce the upstroke of the dose-response curve for CCK-8-stimulated amylase secretion and that very low-affinity CCK receptors mediate the actions of CCK on IP3(1,4,5) and cytosolic calcium and produce the downstroke of the dose-response curve for CCK-8-stimulated amylase secretion. Moreover, CCK-JMV-180 is a full agonist for stimulating amylase secretion by acting at low-affinity CCK receptors and is an antagonist at very low-affinity CCK receptors.

  3. Atividade de amilase em quimo de três espécies tropicais de peixes teleostei de água doce Amylase activity in the chyme of three teleostei freshwater fish

    Directory of Open Access Journals (Sweden)

    José Teixeira de Seixas Filho

    1999-01-01

    Full Text Available O objetivo deste trabalho foi determinar a atividade de amilase no quimo presente nos intestinos médio e posterior, ou reto, em três espécies tropicais de peixes Teleostei de água doce: piracanjuba, Brycon orbignyanus (Valenciennes, 1849, piau, Leporinus friderici (Bloch, 1794, onívoros, e surubim, Pseudoplatystoma coruscans (Agassiz, 1829, preferencialmente carnívoro, visando fornecer referência à nutrição para o ajuste da alimentação artificial. As atividades de amilase foram determinadas usando-se kit BIOCLIN com metodologia modificada por CARAWAY (1959. Os resultados monstraram atividade específica média (2106,33 UA/mg para amilase do piracanjuba e foram 91,74% menor em comparação ao piau (25.488,14 UA/mg, ambos de hábito alimentar onívoro, enquanto a atividade específica da amilase para o piracanjuba foi 89,06% menor em relação ao surubim (19.246,80 UA/mg, carnívoro, o qual apresentou atividade específica da amilase 24,49% menor em relação à do piau. Os dados sugerem que a grande diferença da atividade específica de amilase entre o piau e o piracanjuba possui ligação com sua morfometria e o complexo arranjo das pregas da mucosa dos intestinos médio e posterior. Além disso, a atividade de amilase do surubim indica possibilidade de uso de ração contendo carboidratos, porém estudos adicionais são necessários para a avaliação do comportamento dessas espécies na alimentação artificial.The objective of this work was to determine the amylase activity in the chyme present in the medium and posterior intestines or in the rectum of three tropical freshwater Teleostei species: piracanjuba, Brycon orbignyanus (Valenciennes, 1849, piau, Leporinus friderici (Bloch, 1974, omnivorous, and surubim, Pseudoplatystoma coruscans (Agassiz, 1829, preferably carnivorous, aiming to provide nutritional reference data for the adjustment of the artificial feeding systems. The amylase activity was determined by the use of

  4. U.S. Burning Plasma Organization Activities

    International Nuclear Information System (INIS)

    Fonck, Raymond J.

    2009-01-01

    The national U.S. Burning Plasma Organization (USBPO) was formed to provide an umbrella structure in the U.S. fusion science research community. Its main purpose is the coordination of research activities in the U.S. program relevant to burning plasma science and preparations for participation in the international ITER experiment. This grant provided support for the continuing development and operations of the USBPO in its first years of existence. A central feature of the USBPO is the requirement for broad community participation in and governance of this effort. We concentrated on five central areas of activity of the USBPO during this grant period. These included: (1) activities of the Director and support staff in continuing management and development of the USBPO activity; (2) activation of the advisory Council; (3) formation and initial research activities of the research community Topical Groups; (4) formation of Task Groups to perform specific burning plasma related research and development activities; (5) integration of the USBPO community with the ITER Project Office as needed to support ITER development in the U.S.

  5. Stress-related salivary alpha-amylase (sAA) activity in alcohol dependent patients with and without a history of childhood maltreatment.

    Science.gov (United States)

    Muehlhan, Markus; Höcker, Anja; Höfler, Michael; Wiedemann, Klaus; Barnow, Sven; Schäfer, Ingo

    2017-06-01

    Alcohol-dependent (AD) patients with a history of childhood maltreatment (CM) have shown a more severe clinical profile and a higher risk of relapse than those without CM. It was hypothesized that stress responsivity plays an important role in moderating the relationship between CM and AD. Surprisingly, systematic investigations about the stress responsivity in AD patients with CM are rare. This study compared physiological and subjective stress responses in AD patients with and without CM as well as in healthy controls with and without CM. A total of 130 participants performed the Trier Social Stress Test (TSST). Physiological stress reactivity related to the noradrenergic system was assessed by salivary alpha-amylase (sAA) activity. Subjective ratings of anxiety, nervousness, distress, and mood were rated on visual analogue scales. AD patients showed significantly lower stress-related sAA activity than healthy controls (p ≤ 0.024; z ≥ 1.97). A different pattern was found in the subjective ratings. In particular, anticipatory anxiety revealed a clear effect of CM (p ≤ 0.005; z ≥ 2.43) but no difference between AD patients and healthy controls (p > 0.05). After the TSST, distress ratings differed between AD patients with CM and AD patients without CM (p ≤ 0.009; z ≥ 2.61). The discrepancy between physiological responsivity and subjective stress experiences may account for an increased inability to cope with stressful situations, which in turn might explain the enhanced risk of relapse in AD patients with a history of CM during early abstinence.

  6. Status of plasma physics research activities in Egypt

    International Nuclear Information System (INIS)

    Masoud, M.M.

    1997-01-01

    The status of plasma physics research activities in Egypt is reviewed. There are nine institutes with plasma research activities. The largest is the Atomic energy Authority (AEA), which has activities in fundamental plasma studies, fusion technology, plasma and laser applications, and plasma simulation. The experiments include Theta Pinches, a Z Pinch, a coaxial discharge, a glow discharge, a CO 2 laser, and the EGYPTOR tokamak. (author)

  7. Characterization of salivary alpha-amylase binding to Streptococcus sanguis

    International Nuclear Information System (INIS)

    Scannapieco, F.A.; Bergey, E.J.; Reddy, M.S.; Levine, M.J.

    1989-01-01

    The purpose of this study was to identify the major salivary components which interact with oral bacteria and to determine the mechanism(s) responsible for their binding to the bacterial surface. Strains of Streptococcus sanguis, Streptococcus mitis, Streptococcus mutans, and Actinomyces viscosus were incubated for 2 h in freshly collected human submandibular-sublingual saliva (HSMSL) or parotid saliva (HPS), and bound salivary components were eluted with 2% sodium dodecyl sulfate. By sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western transfer, alpha-amylase was the prominent salivary component eluted from S. sanguis. Studies with 125 I-labeled HSMSL or 125 I-labeled HPS also demonstrated a component with an electrophoretic mobility identical to that of alpha-amylase which bound to S. sanguis. Purified alpha-amylase from human parotid saliva was radiolabeled and found to bind to strains of S. sanguis genotypes 1 and 3 and S. mitis genotype 2, but not to strains of other species of oral bacteria. Binding of [ 125 I]alpha-amylase to streptococci was saturable, calcium independent, and inhibitable by excess unlabeled alpha-amylases from a variety of sources, but not by secretory immunoglobulin A and the proline-rich glycoprotein from HPS. Reduced and alkylated alpha-amylase lost enzymatic and bacterial binding activities. Binding was inhibited by incubation with maltotriose, maltooligosaccharides, limit dextrins, and starch

  8. Evaluation of alpha- amylase inhibition by Urtica dioica and Juglans regia extracts.

    Science.gov (United States)

    Rahimzadeh, Mahsa; Jahanshahi, Samaneh; Moein, Soheila; Moein, Mahmood Reza

    2014-06-01

    One strategy for the treatment of diabetes is inhibition of pancreatic α- amylase. Plants contains different chemical constituents with potential for inhibition of α-amylase and hence maybe used as therapeutic. Urtica dioica and Juglans regia Linn were tested for α-amylase inhibition. Different concentrations of leaf aqueous extracts were incubated with enzyme substrate solution and the activity of enzyme was measured. For determination of the type of inhibition, Dixon plot was depicted. Acarbose was used as the standard inhibitor. Both plant extracts showed time and concentration dependent inhibition of α-amylase. 60% inhibition was seen with 2 mg/ml of U. dioica and 0.4 mg/ml of J. regia aqueous extract. Dixon plots revealed the type of α-amylase inhibition by these two extracts as competitive inhibition. Determination of the type of α-amylase inhibition by these plant extracts could provide by successful use of plant chemicals as drug targets.

  9. Physico-chemical studies on amylases from fermented cassava waste water

    International Nuclear Information System (INIS)

    Oboh, G.; Akindahunsi, A.A.

    2001-09-01

    Waste water from cassava mash fermented with pure strain of Saccharomycees cerevisae together with Lactobacillus delbruckii and Lactobacillus coryneformis (3 days) was assayed for amylase activity. The result of the study indicated that the fermentation waste water had amylase activity, the unit activity and the specific activity of the amylase in the waste water was 0.22μmole/min and 0.06μmole/min/mg, respectively. The amylase was partially purified using Gel filtration (Sephadex-G150). The partially purified enzyme was maximally activity at pH 6.0 and 60 deg. C temperature. It had its maximum stability between pH 6-7 for 4hr, and 30 deg. C for 50 mins. NaCl, NH 4 Cl, FeCl 3 , KCl, NaNO 3 activates the enzyme activity while CUSO 4 and HgCl 2 inhibit the activity of the amylase. It could be concluded that these amylases from the fermented cassava waste amylase were active at wide temperature and pH ranges, this quality could be explored in the industrial sector (most especially food industry) as a source of industrial amylase that requires a wide range of conditions (temperature and pH). (author)

  10. Screening of traditional antidiabetic medicinal plants of Mauritius for possible alpha-amylase inhibitory effects in vitro.

    Science.gov (United States)

    Kotowaroo, M I; Mahomoodally, M F; Gurib-Fakim, A; Subratty, A H

    2006-03-01

    In this study, seven exotic/indigenous medicinal plants of Mauritius, namely Coix lacryma-jobi (Poaceae), Aegle marmelos (Rutaceae), Artocarpus heterophyllus (Moraceae), Vangueria madagascariensis (Rubiaceae), Azadirachta indica (Meliaceae), Eriobotrya japonica (Rosaceae) and Syzigium cumini (Myrtaceae) were studied for possible effects on starch breakdown by alpha-amylase in vitro. The results showed that only Artocarpus heterophyllus significantly (p Artocarpus heterophyllus on alpha-amylase activity using rat plasma in vitro. It was found that the aqueous leaf extract significantly (p Artocarpus heterophyllus behaved as a competitive inhibitor. Results from the present study tend to indicate that Artocarpus heterophyllus could act as a 'starch blocker' thereby reducing post-prandial glucose peaks. Copyright 2006 John Wiley & Sons, Ltd.

  11. Salivary lipase and a-amylase activities are higher in overweight than in normal weight subjects: Influences on dietary beharior

    NARCIS (Netherlands)

    Mennella, I.; Fogliano, V.; Vitaglione, P.

    2014-01-01

    Mounting evidence shows that hedonic eating, leading to overeating just for pleasure, can be driven by oro-sensory factors through the activation of reward processing and learning in the brain. Foods rich in sugars and fats are potent rewards and saliva composition influences oral taste, texture and

  12. Alpha-amylase inhibitor, CS-1036 binds to serum amylase in a concentration-dependent and saturable manner.

    Science.gov (United States)

    Honda, Tomohiro; Kaneno-Urasaki, Yoko; Ito, Takashi; Kimura, Takako; Matsushima, Nobuko; Okabe, Hiromi; Yamasaki, Atsushi; Izumi, Takashi

    2014-03-01

    (2R,3R,4R)-4-hydroxy-2-(hydroxymethyl)pyrrolidin-3-yl 4-O-(6-deoxy-β-D-glucopyranosyl)-α-D-glucopyranoside (CS-1036), which is an α-amylase inhibitor, exhibited biphasic and sustained elimination with a long t1/2 (18.4-30.0 hours) in rats and monkeys, but exhibited a short t1/2 (3.7-7.9 hours) in humans. To clarify the species differences in the t1/2, the plasma protein binding of CS-1036 was evaluated by ultrafiltration. A concentration-dependent and saturable plasma protein binding of CS-1036 was observed in rats and monkeys with the dissociation rate constant (KD) of 8.95 and 27.2 nM, and maximal binding capacity (Bmax) of 52.8 and 22.1 nM, respectively. By the assessments of the recombinant amylase and immunoprecipitation, the major binding protein of CS-1036 in rats was identified as salivary amylase (KD 5.64 nM). CS-1036 also showed concentration-dependent and saturable binding to human salivary and pancreatic amylase, with similar binding affinity in rats. However, the protein binding of CS-1036 was constant in human plasma (≤10.2%) due to the lower serum amylase level compared with rats and monkeys. From the calculation of the unbound fraction (fu) in plasma based on in vitro KD and Bmax, the dose-dependent increase in fu after oral administration is speculated to lead to a dose-dependent increase in total body clearance and a high area under the curve/dose at lower doses, such as 0.3 mg/kg in rats.

  13. Amylase Test: MedlinePlus Lab Test Information

    Science.gov (United States)

    ... page: https://medlineplus.gov/labtests/amylasetest.html Amylase Test To use the sharing features on this page, please enable JavaScript. What is an Amylase Test? An amylase test measures the amount of amylase ...

  14. Biochar activated by oxygen plasma for supercapacitors

    Science.gov (United States)

    Gupta, Rakesh Kumar; Dubey, Mukul; Kharel, Parashu; Gu, Zhengrong; Fan, Qi Hua

    2015-01-01

    Biochar, also known as black carbon, is a byproduct of biomass pyrolysis. As a low-cost, environmental-friendly material, biochar has the potential to replace more expensive synthesized carbon nanomaterials (e.g. carbon nanotubes) for use in future supercapacitors. To achieve high capacitance, biochar requires proper activation. A conventional approach involves mixing biochar with a strong base and baking at a high temperature. However, this process is time consuming and energy inefficient (requiring temperatures >900 °C). This work demonstrates a low-temperature (characteristics are studied. Significant enhancement of the capacitance is achieved: 171.4 F g-1 for a 5-min oxygen plasma activation, in comparison to 99.5 F g-1 for a conventional chemical activation and 60.4 F g-1 for untreated biochar. This enhancement of the charge storage capacity is attributed to the creation of a broad distribution in pore size and a larger surface area. The plasma activation mechanisms in terms of the evolution of the biochar surface and microstructure are further discussed.

  15. Zinc oxide nanoparticles as novel alpha-amylase inhibitors

    Science.gov (United States)

    Dhobale, Sandip; Thite, Trupti; Laware, S. L.; Rode, C. V.; Koppikar, Soumya J.; Ghanekar, Ruchika-Kaul; Kale, S. N.

    2008-11-01

    Amylase inhibitors, also known as starch blockers, contain substances that prevent dietary starches from being absorbed by the body via inhibiting breakdown of complex sugars to simpler ones. In this sense, these materials are projected as having potential applications in diabetes control. In this context, we report on zinc oxide nanoparticles as possible alpha-amylase inhibitors. Zinc oxide nanoparticles have been synthesized using soft-chemistry approach and 1-thioglycerol was used as a surfactant to yield polycrystalline nanoparticles of size ˜18 nm, stabilized in wurtzite structure. Conjugation study and structural characterization have been done using x-ray diffraction technique, Fourier transform infrared spectroscopy, UV-visible spectroscopy, and transmission electron microscopy. Cytotoxicity studies on human fibrosarcoma (HT-1080) and skin carcinoma (A-431) cell lines as well as mouse primary fibroblast cells demonstrate that up to a dose of 20 μg/ml, ZnO nanoparticles are nontoxic to the cells. We report for the first time the alpha-amylase inhibitory activity of ZnO nanoparticles wherein an optimum dose of 20 μg/ml was sufficient to exhibit 49% glucose inhibition at neutral pH and 35 °C temperature. This inhibitory activity was similar to that obtained with acarbose (a standard alpha-amylase inhibitor), thereby projecting ZnO nanoparticles as novel alpha-amylase inhibitors.

  16. Acquired amylase production induced by radiotherapy in a myeloma patient

    Energy Technology Data Exchange (ETDEWEB)

    Yamakawa, Tamami; Nagoshi, Haruhisa; Takahashi, Atushi [Saint Marianna Univ., Kawasaki, Kanagawa (Japan). School of Medicine] [and others

    1995-10-01

    A 55-year-old patient with multiple myeloma (IgG-{lambda}) diagnosed in November 1988 was admitted because of bone pain throughout the body. After plasmapheresis and several courses of chemotherapy, a massive tumor of the left thoracic wall involving the rib appeared. Radiotherapy was performed to ameliorate the severe chest pain, after which myelomatous pleural effusion appeared on the left side. The serum, urine and pleural effusion revealed increased activity of amylase of the salivary type. Amylase activity was also detected in the supernatant of myeloma cells cultured from pleural effusion. We reviewed 12 cases of ectopic amylase-producing multiple myeloma. All the cases except one have been reported from Japan, and hyperamylasemia in these cases was detected at diagnosis or during course of the illness. Moreover, cytogenetic analysis of myeloma cells of previous reports revealed structural abnormalities including chromosome 1, near the amylase gene locus. This case also showed t (1; 10) (q21?; q26) by examination of 8 metaphase derived from bone marrow. These observations suggested that ectopic amylase production was induced by irradiation to the plasmacytoma of thoracic wall. (author).

  17. Acquired amylase production induced by radiotherapy in a myeloma patient

    International Nuclear Information System (INIS)

    Yamakawa, Tamami; Nagoshi, Haruhisa; Takahashi, Atushi

    1995-01-01

    A 55-year-old patient with multiple myeloma (IgG-λ) diagnosed in November 1988 was admitted because of bone pain throughout the body. After plasmapheresis and several courses of chemotherapy, a massive tumor of the left thoracic wall involving the rib appeared. Radiotherapy was performed to ameliorate the severe chest pain, after which myelomatous pleural effusion appeared on the left side. The serum, urine and pleural effusion revealed increased activity of amylase of the salivary type. Amylase activity was also detected in the supernatant of myeloma cells cultured from pleural effusion. We reviewed 12 cases of ectopic amylase-producing multiple myeloma. All the cases except one have been reported from Japan, and hyperamylasemia in these cases was detected at diagnosis or during course of the illness. Moreover, cytogenetic analysis of myeloma cells of previous reports revealed structural abnormalities including chromosome 1, near the amylase gene locus. This case also showed t (1; 10) (q21?; q26) by examination of 8 metaphase derived from bone marrow. These observations suggested that ectopic amylase production was induced by irradiation to the plasmacytoma of thoracic wall. (author)

  18. Interaction of Salivary alpha-Amylase and Amylase-Binding-Protein A (AbpA of Streptococcus gordonii with Glucosyltransferase of S. gordonii and Streptococcus mutans

    Directory of Open Access Journals (Sweden)

    Tanzer Jason M

    2007-06-01

    Full Text Available Abstract Background Glucosyltransferases (Gtfs, enzymes that produce extracellular glucans from dietary sucrose, contribute to dental plaque formation by Streptococcus gordonii and Streptococcus mutans. The alpha-amylase-binding protein A (AbpA of S. gordonii, an early colonizing bacterium in dental plaque, interacts with salivary amylase and may influence dental plaque formation by this organism. We examined the interaction of amylase and recombinant AbpA (rAbpA, together with Gtfs of S. gordonii and S. mutans. Results The addition of salivary alpha-amylase to culture supernatants of S. gordonii precipitated a protein complex containing amylase, AbpA, amylase-binding protein B (AbpB, and the glucosyltransferase produced by S. gordonii (Gtf-G. rAbpA was expressed from an inducible plasmid, purified from Escherichia coli and characterized. Purified rAbpA, along with purified amylase, interacted with and precipitated Gtfs from culture supernatants of both S. gordonii and S. mutans. The presence of amylase and/or rAbpA increased both the sucrase and transferase component activities of S. mutans Gtf-B. Enzyme-linked immunosorbent assay (ELISA using anti-Gtf-B antibody verified the interaction of rAbpA and amylase with Gtf-B. A S. gordonii abpA-deficient mutant showed greater biofilm growth under static conditions than wild-type in the presence of sucrose. Interestingly, biofilm formation by every strain was inhibited in the presence of saliva. Conclusion The results suggest that an extracellular protein network of AbpA-amylase-Gtf may influence the ecology of oral biofilms, likely during initial phases of colonization.

  19. Argania spinosa var. mutica and var. apiculata: variation of fatty-acid composition, phenolic content, and antioxidant and α-amylase-inhibitory activities among varieties, organs, and development stages.

    Science.gov (United States)

    El Adib, Saifeddine; Aissi, Oumayma; Charrouf, Zoubida; Ben Jeddi, Fayçal; Messaoud, Chokri

    2015-09-01

    Argania spinosa includes two varieties, var. apiculata and var. mutica. These argan varieties were introduced into Tunisia in ancient times and are actually cultivated in some botanic gardens. Little is known about the chemical differentiation among these argan varieties. Hence, the aim of this study was to determine the fatty-acid composition, the total phenolic and flavonoid contents, and the antioxidant and α-amylase-inhibitory activities of leaf, seed, and pulp extracts of both argan varieties harvested during the months of January to April. The fatty-acid distribution was found to depend on the argan variety, the plant organ, and the harvest time. Significant variations in the phenolic contents were observed between the investigated varieties as well as between leaves, pulps, and seeds of each variety. As expected, phenolic compounds were found to be contributors to the antioxidant and α-amylase-inhibitory activities of both argan varieties. The chemical differentiation observed among the two argan varieties, based mainly on the fatty-acid composition, might have some chemotaxonomic value. Copyright © 2015 Verlag Helvetica Chimica Acta AG, Zürich.

  20. Amylase production by endophytic fungi Cylindrocephalum sp. isolated from medicinal plant Alpinia calcarata (Haw. Roscoe

    Directory of Open Access Journals (Sweden)

    V. H. Sunitha.

    2012-09-01

    Full Text Available Amylases are among the most important enzymes used in modern biotechnology particularly in the process involving starch hydrolysis. Fungal amylase has large applications in food and pharmaceutical industries. Considering these facts, endophytic fungi isolated from the plant Alpinia calcarata (Haw. Roscoe were screened for amylolytic activity on glucose yeast extract peptone agar (GYP medium. Among thirty isolates of endophytic fungi, isolate number seven identified as Cylindrocephalum sp. (Ac-7 showed highest amylolytic activity and was taken for further study. Influence of various physical and chemical factors such as pH, temperature, carbon and nitrogen sources on amylase production in liquid media were studied. The maximal amylase production was found to be at 30ºC and at pH 7.0 of the growth medium. Among the various carbon and nitrogen sources tested, maltose at 1.5% and Sodium nitrate at 0.3% respectively gave optimum amylase production.

  1. Importance of amylases for physiological quality in maize seeds

    Directory of Open Access Journals (Sweden)

    Camila Aparecida Lopes

    2017-09-01

    Full Text Available Seed quality is the result of the sum of genetic, physical, physiological and sanitary attributes that affect seed ability to perform vital functions related to germination, vigor, and longevity. The expression of genes associated with physiological quality can be assessed by means of germination and vigor analyses, as well as by transcript and protein analyses. The objective in this work was to review the relevance of amylase group enzymes to the physiological quality of maize seeds. Within this group, α-amylase (1,4-α-D-glucan glucanohydrolase E.C 3.2.1.1 plays an important role in starch hydrolysis, and is responsible for 90% of the amylolytic activity in maize seeds. It is responsible for starch conversion into sugars (e.g., destrin, which is used for embryo growth. β-amylase (1,4-α-D-glucan maltohydrolase E.C 3.2.1.2 catalyzes the release of maltose and dextrins from the non-reducing ends of starch. Research has shown that amylase enzymes are directly linked to physiological quality of maize seeds. Alpha- and beta-amylases are mainly involved in the germination process and seed heterosis, and can also be used as molecular markers associated with seed tolerance for drying.

  2. Alpha-amylase from the Hyperthermophilic Archaeon Thermococcus thioreducens

    Science.gov (United States)

    Bernhardsdotter, E. C. M. J.; Pusey, M. L.; Ng, M. L.; Garriott, O. K.

    2003-01-01

    Extremophiles are microorganisms that thrive in, from an anthropocentric view, extreme environments such as hot springs. The ability of survival at extreme conditions has rendered enzymes from extremophiles to be of interest in industrial applications. One approach to producing these extremozymes entails the expression of the enzyme-encoding gene in a mesophilic host such as E.coli. This method has been employed in the effort to produce an alpha-amylase from a hyperthermophile (an organism that displays optimal growth above 80 C) isolated from a hydrothermal vent at the Rainbow vent site in the Atlantic Ocean. alpha-amylases catalyze the hydrolysis of starch to produce smaller sugars and constitute a class of industrial enzymes having approximately 25% of the enzyme market. One application for thermostable alpha-amylases is the starch liquefaction process in which starch is converted into fructose and glucose syrups. The a-amylase encoding gene from the hyperthermophile Thermococcus thioreducens was cloned and sequenced, revealing high similarity with other archaeal hyperthermophilic a-amylases. The gene encoding the mature protein was expressed in E.coli. Initial characterization of this enzyme has revealed an optimal amylolytic activity between 85-90 C and around pH 5.3-6.0.

  3. Salivary Alpha-Amylase Correlates with Subjective Heat Pain Perception.

    Science.gov (United States)

    Wittwer, Amrei; Krummenacher, Peter; La Marca, Roberto; Ehlert, Ulrike; Folkers, Gerd

    2016-06-01

    Self-reports of pain are important for an adequate therapy. This is a problem with patients and infants who are restricted in providing an accurate verbal estimation of their pain. Reliable, real-time, economical, and non-invasive physiological correlates might contribute to a more comprehensive description of pain. Salivary alpha-amylase constitutes one candidate biomarker, which reflects predominantly sympathetic nervous system alterations under stressful conditions and can be measured non-invasively. The current study investigated the effects of acute heat pain on salivary alpha-amylase activity. Heat pain tolerance was measured on the non-dominant forearm. Participants completed visual analog scales on pain intensity and unpleasantness. Saliva samples were collected directly after pain induction. Twenty-seven healthy volunteers were recruited for this study. While salivary alpha-amylase levels correlated positively with intensity and unpleasantness ratings in response to acute heat pain stimuli, there was no corresponding association with pain tolerance. Salivary alpha-amylase is suggested to be an indirect physiologic correlate of subjective heat pain perception. Future studies should address the role of salivary alpha-amylase depending on the origin of pain, the concerned tissue, and other pain assessment methods. © 2016 American Academy of Pain Medicine. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  4. Kinetic studies of acid inactivation of alpha-amylase from Aspergillus oryzae

    DEFF Research Database (Denmark)

    Carlsen, Morten; Nielsen, Jens Bredal; Villadsen, John

    1996-01-01

    The stability of alpha-amylase from Aspergillus oryzae has been studied at different pH. The enzyme is extremely stable at neutral pH (pH 5-8), whereas outside this pH-range a substantial loss of activity is observed. The acid-inactivation of alpha-amylase from A. oryzae was monitored on...... regains part of its activity, and the reactivation process also follows first-order kinetics. The irreversible loss of activity is found not to result from a protease contamination of the protein samples. A proposed model, where irreversibly inactivated a-amylase is formed both directly from the active...

  5. Measurements of amylase isoenzymes in sera and saliva of patients after radiotherapy because of larynx carcinoma

    International Nuclear Information System (INIS)

    Tomasik, A.; Tarnawski, R.; Wierzynski, J.; Tarnawski, R.

    1994-01-01

    Serum and salivary alpha-amylase were measured for controls and patients with laryngeal carcinoma before and after localized irradiation including salivary glands. A significant increase in amylasemia was observed after irradiation. Alpha-amylase activity in saliva was decreased after irradiation but differences were not statistically significant due to the significant decrease of protein in saliva of irradiated group. An increase of salivary isoenzyme S activity was observed while pancreatic isoenzyme activity was not altered. This method allows easy differentiation of hyperamylasemia due to irradiation of parothyroid gland and disorders of the pancreas. Alpha-amylase activity measurements may detect metabolic changes in salivary glands after irradiation. (author)

  6. The Effects of Curcumin on Alpha Amylase in Diabetics Rats

    Directory of Open Access Journals (Sweden)

    Mahmood Najafian

    2015-12-01

    Full Text Available Background One of the therapeutic approaches to lower postprandial blood glucose is to inhibition breakdown of starch by inhibiting carbohydrate hydrolysis enzymes. Alpha-amylase catalyzes the hydrolysis of α-(1, 4-D-glycosidic linkages of starch and other glucose polymers. Inhibitors of this enzyme could be used in the treatment of diabetes. Objectives Based on this purpose we examined the effect of curcumin on alpha amylase and its IC50 and Ki. Materials and Methods In this experimental study, 60 rats were divided into two major groups, normal and diabetic, and each was subsequently divided into five subgroups. One of them as control group that received grape seed oil and four of them as experimental groups that received curcumin at 10, 20, 40 and 80 mg/kg (each group include six rats. Blood glucose levels were measured every three days. Serum insulin levels were measured three times, in the first day, middle and end of the experimental period. The activity of serum alpha amylase was measured in the end of experimental period. Results The results showed that curcumin is a competitive inhibitor for alpha amylase with IC50 = 51.32 µM and Ki = 20.17 µM. In both diabetic and normal groups in all doses nearly dose dependent manner reduced blood glucose and insulin levels. In both diabetic and normal groups decreased levels of serum alpha amylase activity. Conclusions It may be concluded that curcumin is a potent inhibitor of alpha amylase and has beneficial effects in the treatment of overweight and diabetes

  7. Production and Partial Purification of Alpha Amylase from Bacillus subtilis (MTCC 121 Using Solid State Fermentation

    Directory of Open Access Journals (Sweden)

    Dibyangana Raul

    2014-01-01

    Full Text Available Amylase is an enzyme that catalyzes the breakdown of starch into sugars and plays a pivotal role in a variety of areas like use as digestives, for the production of ethanol and high fructose corn syrup, detergents, desiring of textiles, modified starches, hydrolysis of oil-field drilling fluids, and paper recycling. In the present work, solid state fermentation (SSF for α-amylase production has been used in lieu of submerged fermentation (SmF due to its simple technique, low capital investment, lower levels of catabolite repression, and better product recovery. Bacillus subtilis has been well known as producer of alpha amylase and was tested using solid state fermentation for 48 hours at 37°C with wheat bran as substrate. Comparison between different fermentation hours demonstrated high yield of alpha amylase after 48 hours. This alpha amylase has optimum pH and temperature at 7.1 and 40°C, respectively. With the goal to purify alpha amylase, 30–70% (NH42SO4 cut concentrated the amylase activity threefold with respect to crude fermented extract. This was verified in quantitative DNS assay method as well as in zymogram gel profile. The exact molecular weight of the amylase is yet to be determined with the aid of other protein purification techniques.

  8. Mutational analysis of the β-trefoil fold protein barley α-amylase/subtilisin inhibitor probes hot spots for the interaction with barley α-amylase

    DEFF Research Database (Denmark)

    Bønsager, Birgit Christine; Nielsen, P. K.; Abou Hachem, Maher

    2005-01-01

    The barley alpha-amylase/subtilisin inhibitor (BASI) inhibits alpha-amylase 2 (AMY2) with subnanomolar affinity. The contribution of selected side chains of BASI to this high affinity is discerned in this study, and binding to other targets is investigated. Seven BASI residues along the AMY2-BASI...... interface and four residues in the putative protease-binding loop on the opposite side of the inhibitor were mutated. A total of 15 variants were compared with the wild type by monitoring the alpha-amylase and protease inhibitory activities using Blue Starch and azoalbumin, respectively, and the kinetics...

  9. Production and properties of alpha-amylase from Citrobacter species

    Directory of Open Access Journals (Sweden)

    Ebuta N. Etim-Osowo

    2009-04-01

    Full Text Available Amylase production by Citrobacter sp. isolated from potato was optimized in batch culture studies under shake flask conditions. Effects and interactions of best sources and levels of carbon and nitrogen estimated by 4 x 5 and 4 x 4 factorial experimental arrangements were significant (P < 0.01 on amylase production. Optimal alpha-amylase yield was obtained in a medium containing sorghum flour (2.0 % w/v and a mixture of (NH42SO4 + soybean meal (1.5% w/v with an initial medium pH of 8.0. Under optimum conditions, amylase yield was maximal (0.499 U/ml after 60h incubation at room temperature (28oC ± 2oC. Characterization studies showed that the enzyme had maximum activity at 60oC, retained 100% of its original activities at 60oC for 2h, was maximally active at pH 7.0 and retained 100% of original activities at pH 9.0 for 2h. Enzyme activity was stimulated by urea, Mg2+, Ca2+ and Zn2+ but inhibited by Hg2+.

  10. Calcium-induced stabilization of -amylase against guanidine ...

    African Journals Online (AJOL)

    Guanidine hydrochloride (GdnHCl) denaturation of native and Ca- depleted Bacillus licheniformis α-amylase (BLA) was investigated both in the absence and presence of 2 mM calcium chloride (CaCl2) using circular dichroism, fluorescence spectroscopy and biological activity. In both states (Cadepleted and native form), ...

  11. Norepinephrine transporter blocker atomoxetine increases salivary alpha amylase

    NARCIS (Netherlands)

    Warren, C.M.; van den Brink, R.L.; Nieuwenhuis, S.; Bosch, J.A.

    It has been suggested that central norepinephrine (NE) activity may be inferred from increases in salivary alpha-amylase (SAA), but data in favor of this proposition are limited. We administered 40mg of atomoxetine, a selective NE transporter blocker that increases central NE levels, to 24 healthy

  12. Addition of Amylase from Aspergillus Awamori to the Diet of Broiler Chickens

    Directory of Open Access Journals (Sweden)

    HS Morgado

    Full Text Available ABSTRACT Two experiments were performed to evaluate the hematological and blood biochemistry parameters, biometry of digestive organs, enzyme activities, protein content and absolute weight of the pancreas of broilers fed pre-starter and pre-starter diets supplemented or not with amylase from Aspergillus awamori. In total, 120 male Cobb chicks were housed in heated cages in each experiment. A completely randomized experimental design, with two treatments (feed with and without amylase and six replicates per treatment of 10 birds each was applied. The data were subjected to analysis of variance using the F-test at 5% probability level. The dietary amylase addition did not affect hematological and blood biochemistry parameters and the biometry of the gastrointestinal tract of 7- and 21-d-old broilers, nor the absolute weight, enzyme activities or protein concentration of the pancreas of 7-d-old broilers. However, the inclusion of amylase in the diet reduced amylase activity and pancreatic protein concentration in 21-d-old broilers. The application of amylase to broiler chicken pre-starter and starter feeds is not justified given the pancreatic amylase activity and protein concentrations.

  13. Molecular, Biochemical, and Dietary Regulation Features of α-Amylase in a Carnivorous Crustacean, the Spiny Lobster Panulirus argus

    Science.gov (United States)

    Martos-Sitcha, Juan Antonio; Perdomo-Morales, Rolando; Casuso, Antonio; Montero-Alejo, Vivian; García-Galano, Tsai; Martínez-Rodríguez, Gonzalo; Mancera, Juan Miguel

    2016-01-01

    Alpha-amylases are ubiquitously distributed throughout microbials, plants and animals. It is widely accepted that omnivorous crustaceans have higher α-amylase activity and number of isoforms than carnivorous, but contradictory results have been obtained in some species, and carnivorous crustaceans have been less studied. In addition, the physiological meaning of α-amylase polymorphism in crustaceans is not well understood. In this work we studied α-amylase in a carnivorous lobster at the gene, transcript, and protein levels. It was showed that α-amylase isoenzyme composition (i.e., phenotype) in lobster determines carbohydrate digestion efficiency. Most frequent α-amylase phenotype has the lowest digestion efficiency, suggesting this is a favoured trait. We revealed that gene and intron loss have occurred in lobster α-amylase, thus lobsters express a single 1830 bp cDNA encoding a highly conserved protein with 513 amino acids. This protein gives rise to two isoenzymes in some individuals by glycosylation but not by limited proteolysis. Only the glycosylated isoenzyme could be purified by chromatography, with biochemical features similar to other animal amylases. High carbohydrate content in diet down-regulates α-amylase gene expression in lobster. However, high α-amylase activity occurs in lobster gastric juice irrespective of diet and was proposed to function as an early sensor of the carbohydrate content of diet to regulate further gene expression. We concluded that gene/isoenzyme simplicity, post-translational modifications and low Km, coupled with a tight regulation of gene expression, have arose during evolution of α-amylase in the carnivorous lobster to control excessive carbohydrate digestion in the presence of an active α-amylase. PMID:27391425

  14. Molecular, Biochemical, and Dietary Regulation Features of α-Amylase in a Carnivorous Crustacean, the Spiny Lobster Panulirus argus.

    Directory of Open Access Journals (Sweden)

    Leandro Rodríguez-Viera

    Full Text Available Alpha-amylases are ubiquitously distributed throughout microbials, plants and animals. It is widely accepted that omnivorous crustaceans have higher α-amylase activity and number of isoforms than carnivorous, but contradictory results have been obtained in some species, and carnivorous crustaceans have been less studied. In addition, the physiological meaning of α-amylase polymorphism in crustaceans is not well understood. In this work we studied α-amylase in a carnivorous lobster at the gene, transcript, and protein levels. It was showed that α-amylase isoenzyme composition (i.e., phenotype in lobster determines carbohydrate digestion efficiency. Most frequent α-amylase phenotype has the lowest digestion efficiency, suggesting this is a favoured trait. We revealed that gene and intron loss have occurred in lobster α-amylase, thus lobsters express a single 1830 bp cDNA encoding a highly conserved protein with 513 amino acids. This protein gives rise to two isoenzymes in some individuals by glycosylation but not by limited proteolysis. Only the glycosylated isoenzyme could be purified by chromatography, with biochemical features similar to other animal amylases. High carbohydrate content in diet down-regulates α-amylase gene expression in lobster. However, high α-amylase activity occurs in lobster gastric juice irrespective of diet and was proposed to function as an early sensor of the carbohydrate content of diet to regulate further gene expression. We concluded that gene/isoenzyme simplicity, post-translational modifications and low Km, coupled with a tight regulation of gene expression, have arose during evolution of α-amylase in the carnivorous lobster to control excessive carbohydrate digestion in the presence of an active α-amylase.

  15. Molecular, Biochemical, and Dietary Regulation Features of α-Amylase in a Carnivorous Crustacean, the Spiny Lobster Panulirus argus.

    Science.gov (United States)

    Rodríguez-Viera, Leandro; Perera, Erick; Martos-Sitcha, Juan Antonio; Perdomo-Morales, Rolando; Casuso, Antonio; Montero-Alejo, Vivian; García-Galano, Tsai; Martínez-Rodríguez, Gonzalo; Mancera, Juan Miguel

    2016-01-01

    Alpha-amylases are ubiquitously distributed throughout microbials, plants and animals. It is widely accepted that omnivorous crustaceans have higher α-amylase activity and number of isoforms than carnivorous, but contradictory results have been obtained in some species, and carnivorous crustaceans have been less studied. In addition, the physiological meaning of α-amylase polymorphism in crustaceans is not well understood. In this work we studied α-amylase in a carnivorous lobster at the gene, transcript, and protein levels. It was showed that α-amylase isoenzyme composition (i.e., phenotype) in lobster determines carbohydrate digestion efficiency. Most frequent α-amylase phenotype has the lowest digestion efficiency, suggesting this is a favoured trait. We revealed that gene and intron loss have occurred in lobster α-amylase, thus lobsters express a single 1830 bp cDNA encoding a highly conserved protein with 513 amino acids. This protein gives rise to two isoenzymes in some individuals by glycosylation but not by limited proteolysis. Only the glycosylated isoenzyme could be purified by chromatography, with biochemical features similar to other animal amylases. High carbohydrate content in diet down-regulates α-amylase gene expression in lobster. However, high α-amylase activity occurs in lobster gastric juice irrespective of diet and was proposed to function as an early sensor of the carbohydrate content of diet to regulate further gene expression. We concluded that gene/isoenzyme simplicity, post-translational modifications and low Km, coupled with a tight regulation of gene expression, have arose during evolution of α-amylase in the carnivorous lobster to control excessive carbohydrate digestion in the presence of an active α-amylase.

  16. Characterization of purified α-amylase produced by Aspergillus terreus NCFT 4269.10 using pearl millet as substrate

    Directory of Open Access Journals (Sweden)

    Bijay Kumar Sethi

    2016-12-01

    Full Text Available α-amylase was produced by Aspergillus terreus NCFT 4269.10 using both liquid static surface (LSSF and solid-state fermentation using pearl millet residues as substrate. The maximum production of α-amylase was noticed at 30°C incubated for 96h. The crude α-amylase was purified to electrophoretic homogeneity and characterized. Characterization of amylase confirmed that the purified α-amylase was found to be most stable at pH 5.0, 60°C temperature, and a substrate concentration of 1.25%. The enzyme was active for 40 min at 70°C with an optimum enzyme–substrate reaction time of 60 min. Amylase was compatible with all detergents tested having highest activity with Surf excel followed by Henko and Ariel. SDS and Tween 20 reduced the activity. Among the metal ions tested, the maximum α-amylase activity was attained in the presence of Ca2+, followed by Mg2+ and Mn2+. The activity of α-amylase was not considerably affected in the presence of ethylenediaminetetraacetic acid and Triton X-100. Amylase activity was accelerated in the presence of sodium lauryl sulfate and phenylmethylsulfonyl fluoride did not significantly (or slightly affect the activity and stability. Tween 20, urea (5%, and the reducing agent, β-mercaptoethanol significantly inhibited the activity of α-amylase. Owing to its noteworthy stability in the presence of detergents, additives, inhibitors, and metal ions, this α-amylase could be an impending enzyme for significant industrial exploitations.

  17. Plasma Renin Activity in Diabetes Mellitus

    International Nuclear Information System (INIS)

    Pyo, Heui Jung; Park Jung Sik; Kim, Sung Kwon; Choi, Kang Won; Lee, Jung Sang; Lee, Mun Ho

    1979-01-01

    To evaluate the renin-angiotensin-aldosterone system in diabetes mellitus, basal plasma renin activity (PRA) and its response to intravenous furosemide were determined in 40 diabetic subjects. The diabetics were divided into 4 groups according to the presence of nephropathy and/or hypertension. Uncomplicated diabetics (Group I) were taken as control group and the results of the other groups were compared to this group. In diabetics with nephropathy alone (Group II), and with nephropathy and hypertension (Group III), basal PRA values were 0.63±0.59 ng/ml/hr., and 0.79±0.62 ng/ml/hr., respectively, both significantly lower than control group. (1.53±1.09 ng/ml/hr.). (p<0.05) In both of the above groups, the responses to intravenous furosemide tended to be blunted. On the other hand, in diabetics, with hypertension only (Group IV), the basal and stimulated PRA were not significantly different from control. Above results suggests that nephropathy may be one of the factors which suppress renin activity in diabetes mellitus

  18. Plasma Renin Activity in Diabetes Mellitus

    Energy Technology Data Exchange (ETDEWEB)

    Pyo, Heui Jung; Sik, Park Jung; Kim, Sung Kwon; Choi, Kang Won; Lee, Jung Sang; Lee, Mun Ho [Seoul National University College of Medicine, Seoul (Korea, Republic of)

    1979-03-15

    To evaluate the renin-angiotensin-aldosterone system in diabetes mellitus, basal plasma renin activity (PRA) and its response to intravenous furosemide were determined in 40 diabetic subjects. The diabetics were divided into 4 groups according to the presence of nephropathy and/or hypertension. Uncomplicated diabetics (Group I) were taken as control group and the results of the other groups were compared to this group. In diabetics with nephropathy alone (Group II), and with nephropathy and hypertension (Group III), basal PRA values were 0.63+-0.59 ng/ml/hr., and 0.79+-0.62 ng/ml/hr., respectively, both significantly lower than control group. (1.53+-1.09 ng/ml/hr.). (p<0.05) In both of the above groups, the responses to intravenous furosemide tended to be blunted. On the other hand, in diabetics, with hypertension only (Group IV), the basal and stimulated PRA were not significantly different from control. Above results suggests that nephropathy may be one of the factors which suppress renin activity in diabetes mellitus

  19. Liver alpha-amylase gene expression as an early obesity biomarker.

    Science.gov (United States)

    Mojbafan, Marzieh; Afsartala, Zohreh; Amoli, Mahsa M; Mahmoudi, Mahdi; Yaghmaei, Parichehreh; Larijani, Bagher; Ebrahim-Habibi, Azadeh

    2017-04-01

    Obesity is a major health problem worldwide, for which preventive and therapeutic means are still needed. Alpha-amylase is a digestive enzyme whose inhibition has been targeted as a potential anti-obesity strategy. However, alpha-amylase gene expression has not been particularly attended to, and in contrast with pancreatic and salivary amylases, fewer studies have focused on liver alpha-amylase. The present study aimed at investigating the expression of alpha-amylase gene in obese and normal mice at RNA and protein level as well as acarbose effect on this gene expression in hepatocyte cell culture. Control and case groups were fed by normal mouse pellet and high-fat diet respectively, during 8 weeks. After this period, serum biochemical parameters including glucose, cholesterol, triglycerides, AST, ALT and alpha-amylase were assayed. Liver alpha-amylase gene was analyzed by real time PCR, and liver enzyme was assayed with Bernfeld and ELISA methods Hepatocyte cell culture derived from both group were also treated by acarbose and alpha-amylase activity and gene expression was analyzed by above mentioned methods. All biochemical factors showed an increase in obese mice, but the increase in ALT and AST were not statistically significant. Alpha-amylase levels were also increased in obese mice, both at RNA and protein level, while a decrease was seen in obese mice derived hepatocytes after acarbose treatment. Elevated liver alpha-amylase levels may be indicative of initial stages of obesity and the use of acarbose could be considered as a treatment of obesity which could be potentially effective at multiple levels. Copyright © 2016. Published by Elsevier Urban & Partner Sp. z o.o.

  20. Tissue and plasma enzyme activities in juvenile green iguanas.

    Science.gov (United States)

    Wagner, R A; Wetzel, R

    1999-02-01

    To determine activities of intracellular enzymes in 8 major organs in juvenile green iguanas and to compare tissue and plasma activities. 6 green iguanas iguanas, but high values may not always indicate overt muscle disease. The AMS activity may be specific for the pancreas, but the wide range of plasma activity would likely limit its diagnostic usefulness. Activities of AST and LDH may reflect tissue damage or inflammation, but probably do not reflect damage to specific tissues or organs.

  1. Synthesis of the light/pH responsive polymer for immobilization of α-amylase

    Energy Technology Data Exchange (ETDEWEB)

    Yang, Long [Key Laboratory of Applied Surface and Colloid Chemistry, Ministry of Education, School of Chemistry and Chemical Engineering, Shaanxi Normal University, Xi' an 710119 (China); Lei, Ming [School of Material Science and Engineering, Shaanxi Normal University, Xi' an 710119 (China); Zhao, Min [Key Laboratory of Applied Surface and Colloid Chemistry, Ministry of Education, School of Chemistry and Chemical Engineering, Shaanxi Normal University, Xi' an 710119 (China); Yang, Hong [Basic Experimental Teaching Center, Shaanxi Normal University, Xi' an 710062 (China); Zhang, Hong; Li, Yan; Zhang, Kehu [Key Laboratory of Applied Surface and Colloid Chemistry, Ministry of Education, School of Chemistry and Chemical Engineering, Shaanxi Normal University, Xi' an 710119 (China); Lei, Zhongli, E-mail: lzl2016@snnu.edu.cn [Key Laboratory of Applied Surface and Colloid Chemistry, Ministry of Education, School of Chemistry and Chemical Engineering, Shaanxi Normal University, Xi' an 710119 (China)

    2017-02-01

    In this study, light/pH responsive methoxy poly (ethylene glycol)-(5-propargylether-2-nitrobenzyl bromoisobutyrate)-poly methylacrylic acid-b-polystyrene (mPEG-ONB-PMAA-b-PS) polymers were synthesized, and successfully utilized to fabricate micelles and immobilize α-amylase. The critical micelle concentrations (CMC) of the polymers were measured with Pyrene Fluorescent Probe Technique. The morphology and diameter of micelles were characterized by transmission electron microscopy (TEM) and dynamic light scattering (DLS). In addition, the effects of pH, temperature and light-responsive on the catalytic activity were investigated. The optimized fabrication conditions of α-amylase-loaded micelles which α-amylase gave the higher activity were as follows: Immobilization time, 60 min; Immobilization temperature, 50 °C; enzyme concentration, 10 U mL{sup −1}; PBS buffer, pH = 5.4. α-Amylase immobilized in these micelles was much more stable than that free α-amylase. - Highlights: • Light/pH dual-responsive polymer mPEG-ONB-PMAA-b-PS was developed. • The polymer mPEG-ONB-PMAA-b-PS was characterized and utilized to immobilized α-amylase. • A systematic study of dual-responsive polymer influence on α-amylase active was performed.

  2. Capillary electrophoresis as a screening tool for alpha amylase inhibitors in plant extracts

    OpenAIRE

    Hamdan, Imad I.; Afifi, Fatima U.

    2010-01-01

    Capillary electrophoresis (CE) method was developed for screening plant extract for potential alpha amylase (AA) inhibitory activity. The method was validated against a well established UV method. Overall, the proposed method was shown able to detect plants with significant alpha amylase inhibitory activity but not those with rather clinically insignificant activities. Fifty plant species were screened using both the proposed CE method and the UV method and seven plant species were found to p...

  3. Cysteine peroxidase activity in rat blood plasma | Razygraev ...

    African Journals Online (AJOL)

    The rat plasma found to be able to accelerate greatly the H2O2-dependent oxidation of cysteine. The activity was a characteristic of a protein fraction precipitated at 30—44% ammonium sulfate saturation, and the specific activity in protein fraction was significantly higher than in plasma. Cysteine:H2O2 oxidoreductase ...

  4. Raw starch digestion by. alpha. -amylase and glucoamylase from Chalara paradoxa

    Energy Technology Data Exchange (ETDEWEB)

    Monma, Mitsura; Yamamoto, Yoshihiro; Kagei, Norio; Kainuma, Keiji

    1989-10-01

    Glucoamylase and {alpha}-amylase of Chalara paradoxa were separated by hydrophobic column chromatography using butyl-Toyopearl 650M. The {alpha}-amylase showed the highest activity at pH 5.5 and 45{sup 0}C, and was stable in the pH range of 5.5-6.5 and at temperatures lower than 40{sup 0}C. The glucoamylase showed the highest activity at pH 5.0 and 45{sup 0}C, and was stable in the pH range of 4.0-7.5 and at temperatures lower than 45{sup 0}C. The molecular mass of the {alpha}-amylase and glucoamylase estimated by SDS polyacrylamide gel electrophoresis was 80,000 and 68,000, respectively. Both glucoamylase and {alpha}-amylase could digest more effectively raw rice starch and raw corn starch than raw sago starch and raw potato starch. 2% raw rice starch in 10 ml solution was digested by more than 90% by 100 units of each amylase. When these amylases were used combined, raw corn starch was more effectively digested than they were used singly. This cooperative action in raw corn starch digestion was also observed when C. paradoxa {alpha}-amylase and R. niveus glucoamylase were combined. (orig.).

  5. Polymeric amylase nanoparticles as a new semi-synthetic enzyme system for hydrolysis of starch

    Energy Technology Data Exchange (ETDEWEB)

    Say, R. [Anadolu University, Faculty of Science, Chemistry Department, Yunus Emre Campus, Eskişehir (Turkey); Şenay, R. Hilal [Ege University, Faculty of Science, Biochemistry Department, 35100 Bornova-Izmir (Turkey); Biçen, Özlem; Ersöz, Arzu; Şişman Yılmaz, Filiz [Anadolu University, Faculty of Science, Chemistry Department, Yunus Emre Campus, Eskişehir (Turkey); Akgöl, Sinan, E-mail: sinanakgol@yahoo.co.uk [Ege University, Faculty of Science, Biochemistry Department, 35100 Bornova-Izmir (Turkey); Denizli, Adil [Hacettepe University, Faculty of Science, Chemistry Department, 06532 Ankara (Turkey)

    2013-05-01

    α-Amylase (EC 3.2.1.1; α-D-1,4,glucan glucanohydrolase) catalyzes the hydrolysis of α-D-(1,4)-glucosidic linkages in starch, glycogen, and various malto-oligosaccharides, by releasing α-anomeric products. In this study, a novel method has been developed to prepare nanoprotein particles that carry α-amylase as a monomer by using a photosensitive microemulsion polymerization process. The nanostructured α-amylase with photosensitive features have been characterized by fluorescence spectroscopy, transmission electron microscopy (TEM) and Zeta Sizer. The fluorescence intensity of amylase nanoparticles was determined to be 658 a.u. at 610 nm and the average particle size of nanoamylase was found to be about 71.8 nm. Both free α-amylase and nanoparticles were used in the hydrolysis of starch under varying reaction conditions such as pH and temperature that affect enzyme activity and the results were compared to each other. K{sub m} values were 0.26 and 0.87 mM and V{sub max} values were 0.36 IU mg{sup −1} and 22.32 IU mg{sup −1} for nanoenzyme and free enzyme, respectively. Then, thermal stability, storage stability and reusability were investigated and according to the results, activity was preserved 60% at 60 °C; 20% at 70–80 °C temperature values and 80% after 105 days storage. Finally after 10 cycles, the activity was preserved 90% and this novel enzymatic polymeric amylase nanoparticle has showed considerable potential as reusable catalyst. - Highlights: ► Developing to prepare nanoprotein particles carrying α-amylase ► Characterization of nanostructured α-amylase ► Usability of α-amylase nanoparticles in hydrolysis of starch.

  6. Polymeric amylase nanoparticles as a new semi-synthetic enzyme system for hydrolysis of starch

    International Nuclear Information System (INIS)

    Say, R.; Şenay, R. Hilal; Biçen, Özlem; Ersöz, Arzu; Şişman Yılmaz, Filiz; Akgöl, Sinan; Denizli, Adil

    2013-01-01

    α-Amylase (EC 3.2.1.1; α-D-1,4,glucan glucanohydrolase) catalyzes the hydrolysis of α-D-(1,4)-glucosidic linkages in starch, glycogen, and various malto-oligosaccharides, by releasing α-anomeric products. In this study, a novel method has been developed to prepare nanoprotein particles that carry α-amylase as a monomer by using a photosensitive microemulsion polymerization process. The nanostructured α-amylase with photosensitive features have been characterized by fluorescence spectroscopy, transmission electron microscopy (TEM) and Zeta Sizer. The fluorescence intensity of amylase nanoparticles was determined to be 658 a.u. at 610 nm and the average particle size of nanoamylase was found to be about 71.8 nm. Both free α-amylase and nanoparticles were used in the hydrolysis of starch under varying reaction conditions such as pH and temperature that affect enzyme activity and the results were compared to each other. K m values were 0.26 and 0.87 mM and V max values were 0.36 IU mg −1 and 22.32 IU mg −1 for nanoenzyme and free enzyme, respectively. Then, thermal stability, storage stability and reusability were investigated and according to the results, activity was preserved 60% at 60 °C; 20% at 70–80 °C temperature values and 80% after 105 days storage. Finally after 10 cycles, the activity was preserved 90% and this novel enzymatic polymeric amylase nanoparticle has showed considerable potential as reusable catalyst. - Highlights: ► Developing to prepare nanoprotein particles carrying α-amylase ► Characterization of nanostructured α-amylase ► Usability of α-amylase nanoparticles in hydrolysis of starch

  7. Isolation, characterization and partial purification of alpha-amylase from a marine bacillus NH-25

    International Nuclear Information System (INIS)

    Ahmad, M.; Zohra, R.R.

    2012-01-01

    Total 399 marine strains were isolated from the sea water sample and screened for thermostable amylase production. Out of these 52 were to have amylogenic activity. Among them 2 isolates were able to grow and produce amylase at 55 degree C. Strain NH-25 tolerates 30% salt, a wide j-H range (4-8) and retained 64% activity at 50 degree C after 60 minutes. (author)

  8. Effect of alpha amylase on early childhood caries: a matched case-control study

    OpenAIRE

    Mojarad, Farzad; Department of Pediatric Dentistry, School of Dentistry, Hamadan University of Medical Sciences, Hamadan; Fazlollahifar, Samira; Department of Pediatric Dentistry, School of Dentistry, Hamadan University of Medical Sciences, Hamadan; Poorolajal, Jalal; Research Center for Health Sciences, Department of Epidemiology & Biostatistics, School of Public Health, Hamadan University of Medical Sciences, Hamadan; Hajilooi, Mehrdad; Department of Pathology, School of Dentistry, Hamadan University of Medical Sciences, Hamadan

    2013-01-01

    Objectives: There are a few studies addressing the relationship between salivary alpha-amylase and dental caries. This study was implemented in order to investigate the effect of salivary alpha-amylase level on early childhood caries (ECC).Materials and Methods: In this matched case-control study, which was carried out from November 2011 to March 2012 in Hamadan City, the west of Iran, mean levels of salivary alpha-amylase of 84 ECC-active cases were compared to that of 84 ECC-free controls u...

  9. ISOLATION AND IDENTIFICATION OF AMYLASE PRODUCING YEASTS IN ‘TELLA’ (ETHIOPIAN LOCAL BEER AND THEIR AMYLASE CONTRIBUTION FOR ‘TELLA’ PRODUCTION

    Directory of Open Access Journals (Sweden)

    Berhanu Andualem

    2013-08-01

    Full Text Available ‘Tella’ is local beer which is used in most part of Ethiopia. It is made from cereals, such as barley, wheat, maize and other crops. Rhamnus prinoides is also used to provide a special aroma and flavor as well as antiseptic agent. The objective of this study is to determine the contribution of amylases from tella yeast isolates and compare with the role of amylase from malt. House hold ‘tella’ samples were collected and plated on starch agar and then amylase positive isolates of yeast were identified by folding iodine solution over the starch agar. Amylase assay and activities were investigated by standard methods and compared with amylase from malt. According to this study, the activity of amylases which was extracted from yeast isolates was very low and may have no contribution in the conversion of starch into fermentable sugars. Thus, it is better to avoid such organisms from ‘tella’ fermentation in order to discriminate unwanted bio-products. In conclusion, the substrates and ingredients should be sterilized and introduced into the fermentation system aseptically.

  10. The effects of a three-week resistance exercise plus whole body vibration (WBV on the testosterone-free cortisole ratio differences and activity of the α-salivary amylase enzyme

    Directory of Open Access Journals (Sweden)

    MohammadAli Fathi

    2013-01-01

    Full Text Available Background and Aim: Researchers have already reported the hormonal responses of the effects of isolated resistance exercise or whole body vibration (WBV. However, no documents are available on the combination of the resistance exercise plus WBV on biomarkers, particularly salivary α-amylase activity as well as free testosterone to cortisol ratio (FTCR. Materials and Methods: To discover this, thirty young football players were randomly allocated into three WBV (n=10, resistance exercise (n=10, and a combination of both (n=10. The subjects took nine sessions of exercise during a three-week period. The WBV included one minute exercise followed by one minute rest for 10 minutes with 50 Hz frequency, four mm. transverse translation and in semi-squat standing condition. The resistive exercise included three-period exercises with 70% one-maximal repetition with a two-minute rest interval. The third group carried out the resistance exercise firstly and then WBV. The pre / post salivary samples were collected. Results: It showed that the concentration of the cortisol and the salivary α-amylaseactivity increased significantly. The testosterone increased among all groups following the first session, but significantly decreased at the last session. The statistical analysis showed a significant increase in cortisol concentration only in WBV group (p=0.004. Conclusion: The results confirmed that hormone secretion requires an acceptable threshold of both intensity and duration of training. In addition, secretion of the α-salivary amylase responses to various physiologic stresses immediately. In submaximal exercises, there is probably no difference among training techniques, WHB and strengthening exercises. The current study also emphasizes on salivary α-amylaseactivity immediately following physiological stresses.

  11. Two-step purification and partial characterization of an extra cellular α-amylase from Bacillus licheniformis

    Directory of Open Access Journals (Sweden)

    Zare Mirakabadi, A.

    2012-11-01

    Full Text Available The aim of this study was production and partial purification of α-amylase enzyme by Bacillus licheniformis. B. Licheniformis was allowed to grow in broth culture for purpose of inducing α-amylase enzyme. Optimal conditions for amylase production by B. Licheniformis are incubation period of 120 h, temperature of 37 °C and pH 7.0. The α-amylase enzyme was purified by ion exchange chromatography on DEAE-sepharose CL-6B and sephadex G-100 gel filtration with a 19.1-fold increase in specific activity as compared to the concentrated supernatant and with a specific activity of 926.47 U/mg. The α-amylase had the highest activity at pH 7.0 and 65 °C. According to the data on native polyacrylamide gel electrophoresis, the molecular weight of the purified enzyme was 72 kDa.

  12. Plasma Chamber Design and Fabrication Activities

    Science.gov (United States)

    Parodi, B.; Bianchi, A.; Cucchiaro, A.; Coletti, A.; Frosi, P.; Mazzone, G.; Pizzuto, A.; Ramogida, G.; Coppi, B.

    2006-10-01

    A fabrication procedure for a typical Plasma Chamber (PC) sector has been developed to cover all the manufacturing phases, from the raw materials specification (including metallurgical processes) to the machining operations, acceptance procedures and vacuum tests. Basically, the sector is made of shaped elements (forged or rolled) welded together using special fixtures and then machined to achieve the final dimensional accuracy. An upgraded design of the plasma chamber's vertical support that can withstand the estimated electromagnetic loads (Eddy and Halo current plus horizontal net force resulting from the worst plasma disruption scenario VDE, Vertical Displacement Event) has been completed. The maintenance of the radial support can take place hands-on with a direct access from outside the cryostat. With the present design, vacuum tightness is achieved by welding conducted with automatic welding heads. On the outer surface of the PC a dedicated duct system, filled by helium gas, is included to cool down the PC to room temperature when needed.

  13. Plasma dihydroxyphenylalanine (DOPA) is independent of sympathetic activity in humans

    DEFF Research Database (Denmark)

    Eldrup, E; Christensen, N J; Andreasen, J

    1989-01-01

    in diabetic patients with autonomic neuropathy compared to diabetics without neuropathy, whereas baseline plasma DOPA concentrations were similar in the three groups investigated: 6.55 (5.03-7.26, median [interquartile range], n = 8) nmol l-1 in diabetics with neuropathy, 7.41 (5.79-7.97, n = 8) nmol l-1...... in diabetics without neuropathy, and 6.85 (5.58-7.36, n = 8) nmol l-1 in controls. No relationship was obtained between baseline values of plasma NE and plasma DOPA. Plasma DOPA did not change in the upright position, whereas plasma NE increased significantly. Our results indicate that plasma DOPA...... is not related to sympathetic activity and may be of non-neuronal origin....

  14. α-Amylase inhibitors: a review of raw material and isolated compounds from plant source.

    Science.gov (United States)

    Sales, Paloma Michelle; Souza, Paula Monteiro; Simeoni, Luiz Alberto; Silveira, Damaris

    2012-01-01

    Inhibition of α-amylase, enzyme that plays a role in digestion of starch and glycogen, is considered a strategy for the treatment of disorders in carbohydrate uptake, such as diabetes and obesity, as well as, dental caries and periodontal diseases. Plants are an important source of chemical constituents with potential for inhibition of α-amylase and can be used as therapeutic or functional food sources. A review about crude extracts and isolated compounds from plant source that have been tested for α-amylase inhibitory activity has been done. The analysis of the results shows a variety of crude extracts that present α-amylase inhibitory activity and some of them had relevant activity when compared with controls used in the studies. Amongst the phyto-constituents that have been investigated, flavonoids are one of them that demonstrated the highest inhibitory activities with the potential of inhibition related to number of hydroxyl groups in the molecule of the compound. Several phyto-constituents and plant species as α-amylase inhibitors are being reported in this article. Majority of studies have focused on the anti-amylase phenolic compounds.

  15. Differences in muscle pain and plasma creatine kinase activity after ...

    African Journals Online (AJOL)

    encephalopathy,18 and the decrement in muscle power associated with muscle damage.6 ... A high degree of intra-individual variability in plasma. CK activity was ..... 21. Komi PV. Stretch-shortening cycle exercise: a powerful model to study.

  16. Biology-oriented drug synthesis (BIODS) of 2-(2-methyl-5-nitro-1H-imidazol-1-yl)ethyl aryl ether derivatives, in vitro α-amylase inhibitory activity and in silico studies.

    Science.gov (United States)

    Taha, Muhammad; Imran, Syahrul; Ismail, Nor Hadiani; Selvaraj, Manikandan; Rahim, Fazal; Chigurupati, Sridevi; Ullah, Hayat; Khan, Fahad; Salar, Uzma; Javid, Muhammad Tariq; Vijayabalan, Shantini; Zaman, Khalid; Khan, Khalid Mohammed

    2017-10-01

    A new library of 2-(2-methyl-5-nitro-1H-imidazol-1-yl)ethyl aryl ether derivatives (1-23) were synthesized and characterized by EI-MS and 1 H NMR, and screened for their α-amylase inhibitory activity. Out of twenty-three derivatives, two molecules 19 (IC 50 =0.38±0.82µM) and 23 (IC 50 =1.66±0.14µM), showed excellent activity whereas the remaining compounds, except 10 and 17, showed good to moderate inhibition in the range of IC 50 =1.77-2.98µM when compared with the standard acarbose (IC 50 =1.66±0.1µM). A plausible structure-activity relationship has also been presented. In addition, in silico studies was carried out in order to rationalize the binding interaction of compounds with the active site of enzyme. Copyright © 2017 Elsevier Inc. All rights reserved.

  17. Determination of activated plasma fibronectin using radioactive labelled collagen I

    DEFF Research Database (Denmark)

    Fenger, M

    1984-01-01

    The plasma concentration of biological active fibronectin was assayed by a protein binding assay using 125I-collagen I as ligand and heparin as activator. The standard curve is linear for a fibronectin range of 1.1-11 pmol (0.5-5.0 micrograms) and the coefficient of variation was less than 10......%. The active or activable fibronectin was compared to the immunoreactive fibronectin in plasma from patients with various bacterial diseases. Similar concentrations were detected by the two assays suggesting that all the circulating fibronectin was functionally active. The assay was also applied to determine...

  18. Plasma technology of the surface polymer activation

    International Nuclear Information System (INIS)

    Dutra, Jorge C.N.; Mello, Sandra C.; Massi, Marcos; Otani, Choyu; Maciel, Homero S.; Bittencourt, Edison

    2005-01-01

    A number of polymers, especially rubbers, require surface treatment to achieve a satisfactory level of adhesion. The surface of EPDM rubber vulcanized is high hydrophobicity and is not suited for a number of potential applications, in particular, for adhering to the polyurethane liner of solid rocket propellants. In this case, plasma treatment can be a very attractive process because it can efficiently increase the surface energy attributed to surface oxidation with the introduction of polar groups 1, 2. In order to investigate the influence of the parameters on the modifications of the treated surface samples of EPDM rubber by plasma generated by gas oxygen and argon, the water and methylene iodide contact angles were measured at room temperature with an image analyzing using the sessile drop technique 3 - 6 . (author)

  19. Physical and catalytic properties of alpha-amylase from Tenebrio molitor L. larvae.

    Science.gov (United States)

    Buonocore, V; Poerio, E; Silano, V; Tomasi, M

    1976-01-01

    The amylase from Tenebrio molitor L. larvae (yellow mealworm) was characterized according to a number of its molecular and catalytic properties. The insect amylase is a single polypeptide chain with mol.wt. 68000, an isoelectric point of 4.0 and a very low content of sulphur-containing amino acids. The enzyme is a Ca2+-protein and behaves as an alpha-amylase. Removal of Ca2+ by exhaustive dialysis against water causes the irreversible inactivation of the enzyme. Moreover, the enzyme is activated by the presence in the assay mixture of Cl-, or some other inorganic anions that are less effective than Cl-, and is inhibited by F-. Optimal conditions of pH and temperature for the enzymic activity are 5.8 and 37 degrees C. The insect amylase exhibits an identical kinetic behaviour toward starch, amylose and amylopectin; the enzyme hydrolyses glycogen with a higher affinity constant. Compared with the non-insect alpha-amylases described in the literature, Tenebrio molitor amylase has a lower affinity for starch. PMID:942374

  20. Production of Amylase by Bacillus polymyxa NCIM No. 2539 from Agroindustrial Wastes

    Directory of Open Access Journals (Sweden)

    Abhishek Dutt Tripathi

    2017-04-01

    Full Text Available Background and Objective: In the present study, Bacillus polymyxa NCIM No. 2539 was selected to utilize agro-industrial byproduct (orange peel for amylase production under submergedfermentation conditions.Material and Methods: Different agro-industrial byproducts like cane molasses, wheat bran, rice bran and orange peel were screened for maximum amylase production. Amylase activitiy of Bacillus polymyxa was studied using starch-agar plate method. MINITAB software Version 17 and central composite design were applied to evaluate effect of supplementation of substrate with different sulphur containing amino acids (cysteine, methionine and cystine and vitamin thiamine on enzyme activity. Further optimization of the parameters viz. amount of substrate, concentration of amino acid and vitamin for maximum amylase production was studied by central composite rotatable design.Results and Conclusion: Among 4 different agro-industrial substrates applied, orange peel showed maximum enzyme production (activity: 492.31 IU g-1 sample. Supplementation of the production media with cysteine showed maximum amylaseproduction (515.38 IU g-1 sample among all three amino acids and control. Supplementation with thiamine also showed more amylase production (469.23 IU g-1 sample as compared to control (415.38 IU g-1. Cysteine and thiamine proved to increaseamylase production significantly. Maximum amylase production was obtained at 7.7 g orange peel, 37.29 mg cysteine and 34.23 mg per 10 ml thiamine.Conflict of interest: The authors declare no conflict of interest.

  1. Isolation and characterization of α-amylase from marine ...

    African Journals Online (AJOL)

    The α-amylase of marine Pseudomonas sp. K6-28-040 was purified through a series of three steps and the purity of enzymes was checked by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The results showed that, the enzyme was purified 4.7-fold with a specific activity of 134.6 U/mg protein and a yield of 44% ...

  2. Biotechnological Processes in Microbial Amylase Production.

    Science.gov (United States)

    Gopinath, Subash C B; Anbu, Periasamy; Arshad, M K Md; Lakshmipriya, Thangavel; Voon, Chun Hong; Hashim, Uda; Chinni, Suresh V

    2017-01-01

    Amylase is an important and indispensable enzyme that plays a pivotal role in the field of biotechnology. It is produced mainly from microbial sources and is used in many industries. Industrial sectors with top-down and bottom-up approaches are currently focusing on improving microbial amylase production levels by implementing bioengineering technologies. The further support of energy consumption studies, such as those on thermodynamics, pinch technology, and environment-friendly technologies, has hastened the large-scale production of the enzyme. Herein, the importance of microbial (bacteria and fungi) amylase is discussed along with its production methods from the laboratory to industrial scales.

  3. Biotechnological Processes in Microbial Amylase Production

    Directory of Open Access Journals (Sweden)

    Subash C. B. Gopinath

    2017-01-01

    Full Text Available Amylase is an important and indispensable enzyme that plays a pivotal role in the field of biotechnology. It is produced mainly from microbial sources and is used in many industries. Industrial sectors with top-down and bottom-up approaches are currently focusing on improving microbial amylase production levels by implementing bioengineering technologies. The further support of energy consumption studies, such as those on thermodynamics, pinch technology, and environment-friendly technologies, has hastened the large-scale production of the enzyme. Herein, the importance of microbial (bacteria and fungi amylase is discussed along with its production methods from the laboratory to industrial scales.

  4. Plasma renin activity profile in normal and hypertensive Filipinos

    International Nuclear Information System (INIS)

    Guevara, R.; Torres, J. Jr.; Abundo, H.P.; Perez, A.P.

    To establish a base line profile of plasma renin activity in normotensive and hypertensive Filipinos, 1.019 cases, 479 males and 540 females with an age range 14 - 89 years (mean - 46 + -20) were studied at the Santo Tomas University Hospital of various life styles from the Metro-Manila area, 248 comprised the normotensive group (male - 122 or 49.2 %, female 126 or 50.8 %) and 771 were hypertensive. Of these, 711 (92.6 %) has essential hypertension and are presented in this report. Plasma Renin Activity was determined by radioimmunoassay using Dainabot Renin-Ricket. Concurrent 24 hr. urine sodium and potassium were determined. Nomograms of plasma renin activity as related to urine sodium excretion were drawn after computerized statistical analysis of data. The normal mean value of plasma renin activity was found to be 1.64 + - 0.81 ng./ml./hr. in the upright position and 1.15 + - .68 ng./ml./hr. in the supine position. Based on the nomogram derived, the values obtained in the 711 cases of essential hypertension were classified into High Renin - 14.3 % Normal Renin - 56.1 % and Low Renin - 29.6 %. This study establishes normal levels of plasma renin activity as well as define and classify same renin activity among hypertensive Filipinos, a useful and practicable guide for treatment and can be of prognostic significance. (author)

  5. Correlation between rivaroxaban (Xarelto) plasma activity, patient ...

    African Journals Online (AJOL)

    with resultant marked fluctuation in anticoagulation control in a significant number ... activity, patient clinical variables and outcomes ... clinical laboratories to have the ability to measure .... To assess pharmacological activity, i.e. quality control.

  6. Active neutral particle diagnostics for high temperature plasma

    International Nuclear Information System (INIS)

    Tobita, Kenji

    1993-01-01

    This paper describes experimental studies related to active neutral particle diagnostics in the JT-60 tokamak. Detection efficiencies of a micro-channel plate (MCP), which has widely used in plasma diagnostics, were determined for ions and neutrals. Multi-step processes for a neutral beam is predicted to enhance the beam stopping cross section in a plasma. In order to confirm the predictions, shine-through for a hydrogen and for a helium beam was measured in the JT-60 ohmic plasmas. The measurements for a hydrogen beam resulted in the cross sectional enhancement in the beam stopping. The same experiment using a helium beam indicated that the cross sectional enhancement for helium was much smaller than that for hydrogen at almost same plasma parameters. Ion temperature diagnostic using active beam scattering was developed in data processing technique, in consideration of the device function of a neutral particle analyzer and in estimation of the effect of beam ion component. Fundamental experiments for detecting helium ions in a plasma were performed using two-electron transfer reaction between a helium atomic beam and helium ions, and the energy distribution and the density of the helium ions were determined. These experiments demonstrated promise of the two-electron transfer reaction as an alpha ash detection in a burning plasma. A parasitic neutral efflux accompanied by active beam injection was investigated. (J.P.N.)

  7. POTENTIAL USE OF AN EXTRACELLULAR ENZYME OF a-AMYLASE FROM INDIGENOUS INDONESIAN MESOPHILIC BACTERIA

    Directory of Open Access Journals (Sweden)

    Puji Lestari

    2013-04-01

    Full Text Available Amylase enzyme has a great significance for industrial usages in  Indonesia. However, this enzyme is still imported. The use of bacteria in biotechnological process of industrial products such as enzyme production has stimulated the exploration of extracellular amylase producing  bacteria. This study aimed to identify and analyze the potential use of amylolytic bacterial enzymes for hydrolyzing cassava starch. Two bacterial isolates, i.e. MII-10 and DKW-8 originated from Indonesia soil were identified based on their morphological, physiological and biochemical properties according to the standard protocol. The isolates were then  cultivated on fermentation medium and their growth pattern and  enzymatic assays were observed. The acetone-precipitated crude enzyme harvested based on predetermined cultivation time was used for  enzymatic hydrolysis product characterization on cassava starch using thin layer chromatography (TLC. The results showed that the mesophilicbacteria isolates (MII-10 and DKW-8 were belonged to Bacillus licheniformis. The maximum bacterial cell growth and enzyme activity were reached at 48 hours after incubation. The MII-10 isolate was found more stable than DKW-8 in producing amylase enzyme. Amylase produced by the MII-10 and DKW- 8 isolates was identified to be an endo-a-amylase as confirmed by oligosaccharides and dextrin of the random hydrolysisproducts. Relatively high dextrose equivalence (DE value of a-amylase of MII-10 (DE of 9.96 suggests that the enzyme is prospective for  saccharification of starchy material in glucose syrup industry.

  8. Evaluation of alpha- amylase inhibition by Urtica dioica and Juglans regia extracts

    Directory of Open Access Journals (Sweden)

    Mahsa Rahimzadeh

    2014-06-01

    Full Text Available Objective(s:One strategy for the treatment of diabetes is inhibition of pancreatic α- amylase. Plants contains different chemical constituents with potential for inhibition of α-amylase and hence maybe used as therapeutic. Materials and Methods: Urtica dioica and Juglans regia Linn were tested for α-amylase inhibition. Different concentrations of leaf aqueous extracts were incubated with enzyme substrate solution and the activity of enzyme was measured. For determination of the type of inhibition, Dixon plot was depicted. Acarbose was used as the standard inhibitor. Results: Both plant extracts showed time and concentration dependent inhibition of α-amylase. 60% inhibition was seen with 2 mg/ml of U. dioica and0.4 mg/ml of J. regia aqueous extract. Dixon plots revealed the type of α-amylase inhibition by these two extracts as competitive inhibition. Conclusion: Determination of the type of α-amylase inhibition by these plant extracts could provide by successful use of plant chemicals as drug targets.

  9. Synthesis and study of the α-amylase inhibitory potential of thiadiazole quinoline derivatives.

    Science.gov (United States)

    Taha, Muhammad; Tariq Javid, Muhammad; Imran, Syahrul; Selvaraj, Manikandan; Chigurupati, Sridevi; Ullah, Hayat; Rahim, Fazal; Khan, Fahad; Islam Mohammad, Jahidul; Mohammed Khan, Khalid

    2017-10-01

    α-Amylase is a target for type-2 diabetes mellitus treatment. However, small molecule inhibitors of α-amylase are currently scarce. In the course of developing small molecule α-amylase inhibitors, we designed and synthesized thiadiazole quinoline analogs (1-30), characterized by different spectroscopic techniques such as 1 HNMR and EI-MS and screened for α-amylase inhibitory potential. Thirteen analogs 1, 2, 3, 4, 5, 6, 22, 23, 25, 26, 27, 28 and 30 showed outstanding α-amylase inhibitory potential with IC 50 values ranges between 0.002±0.60 and 42.31±0.17μM which is many folds better than standard acarbose having IC 50 value 53.02±0.12μM. Eleven analogs 7, 9, 10, 11, 12, 14, 15, 17, 18, 19 and 24 showed good to moderate inhibitory potential while seven analogs 8, 13, 16, 20, 21 and 29 were found inactive. Our study identifies novel series of potent α-amylase inhibitors for further investigation. Structure activity relationship has been established. Copyright © 2017 Elsevier Inc. All rights reserved.

  10. Determination of activated plasma fibronectin using radioactive labelled collagen I

    DEFF Research Database (Denmark)

    Fenger, M

    1984-01-01

    The plasma concentration of biological active fibronectin was assayed by a protein binding assay using 125I-collagen I as ligand and heparin as activator. The standard curve is linear for a fibronectin range of 1.1-11 pmol (0.5-5.0 micrograms) and the coefficient of variation was less than 10...

  11. Plasma Renin Activity in Children with Protein Energy Malnutrition ...

    African Journals Online (AJOL)

    Plasma renin activity was measured by bio-assay in 100 children with kwashiorkor and in 20 healthy children, and also by radio-immunoassay in another 26 children with kwashiorkor and in another 20 healthy children. Both methods showed that (compared with healthy children) renin activity was significantly increased in ...

  12. Development of active porous medium filters based on plasma textiles

    International Nuclear Information System (INIS)

    Kuznetsov, Ivan A.; Saveliev, Alexei V.; Rasipuram, Srinivasan; Kuznetsov, Andrey V.; Brown, Alan; Jasper, Warren

    2012-01-01

    Inexpensive, flexible, washable, and durable materials that serve as antimicrobial filters and self-decontaminating fabrics are needed to provide active protection to people in areas regularly exposed to various biohazards, such as hospitals and bio research labs working with pathogens. Airlines and cruise lines need such material to combat the spread of infections. In households these materials can be used in HVAC filters to fight indoor pollution, which is especially dangerous to people suffering from asthma. Efficient filtering materials are also required in areas contaminated by other types of hazardous dust particulates, such as nuclear dust. The primary idea that guided the undertaken study is that a microplasma-generating structure can be embedded in a textile fabric to generate a plasma sheath (''plasma shield'') that kills bacterial agents coming in contact with the fabric. The research resulted in the development of a plasma textile that can be used for producing new types of self-decontaminating garments, fabrics, and filter materials, capable of activating a plasma sheath that would filter, capture, and destroy any bacteriological agent deposited on its surface. This new material relies on the unique antimicrobial and catalytic properties of cold (room temperature) plasma that is benign to people and does not cause thermal damage to many polymer textiles, such as Nomex and polypropylene. The uniqueness of cold plasma as a disinfecting agent lies in the inability of bacteria to develop resistance to plasma exposure, as they can for antibiotics. Plasma textiles could thus be utilized for microbial destruction in active antimicrobial filters (for continuous decontamination and disinfection of large amounts of air) as well as in self-decontaminating surfaces and antibacterial barriers (for example, for creating local antiseptic or sterile environments around wounds and burns).

  13. Development of active porous medium filters based on plasma textiles

    Science.gov (United States)

    Kuznetsov, Ivan A.; Saveliev, Alexei V.; Rasipuram, Srinivasan; Kuznetsov, Andrey V.; Brown, Alan; Jasper, Warren

    2012-05-01

    Inexpensive, flexible, washable, and durable materials that serve as antimicrobial filters and self-decontaminating fabrics are needed to provide active protection to people in areas regularly exposed to various biohazards, such as hospitals and bio research labs working with pathogens. Airlines and cruise lines need such material to combat the spread of infections. In households these materials can be used in HVAC filters to fight indoor pollution, which is especially dangerous to people suffering from asthma. Efficient filtering materials are also required in areas contaminated by other types of hazardous dust particulates, such as nuclear dust. The primary idea that guided the undertaken study is that a microplasma-generating structure can be embedded in a textile fabric to generate a plasma sheath ("plasma shield") that kills bacterial agents coming in contact with the fabric. The research resulted in the development of a plasma textile that can be used for producing new types of self-decontaminating garments, fabrics, and filter materials, capable of activating a plasma sheath that would filter, capture, and destroy any bacteriological agent deposited on its surface. This new material relies on the unique antimicrobial and catalytic properties of cold (room temperature) plasma that is benign to people and does not cause thermal damage to many polymer textiles, such as Nomex and polypropylene. The uniqueness of cold plasma as a disinfecting agent lies in the inability of bacteria to develop resistance to plasma exposure, as they can for antibiotics. Plasma textiles could thus be utilized for microbial destruction in active antimicrobial filters (for continuous decontamination and disinfection of large amounts of air) as well as in self-decontaminating surfaces and antibacterial barriers (for example, for creating local antiseptic or sterile environments around wounds and burns).

  14. Development of active porous medium filters based on plasma textiles

    Energy Technology Data Exchange (ETDEWEB)

    Kuznetsov, Ivan A.; Saveliev, Alexei V.; Rasipuram, Srinivasan; Kuznetsov, Andrey V.; Brown, Alan; Jasper, Warren [Mechanical and Aerospace Engineering, North Carolina State University, Raleigh, NC 27695 (United States); Textile Engineering Chemistry and Science, North Carolina State University, Raleigh, NC 27695 (United States)

    2012-05-15

    Inexpensive, flexible, washable, and durable materials that serve as antimicrobial filters and self-decontaminating fabrics are needed to provide active protection to people in areas regularly exposed to various biohazards, such as hospitals and bio research labs working with pathogens. Airlines and cruise lines need such material to combat the spread of infections. In households these materials can be used in HVAC filters to fight indoor pollution, which is especially dangerous to people suffering from asthma. Efficient filtering materials are also required in areas contaminated by other types of hazardous dust particulates, such as nuclear dust. The primary idea that guided the undertaken study is that a microplasma-generating structure can be embedded in a textile fabric to generate a plasma sheath (''plasma shield'') that kills bacterial agents coming in contact with the fabric. The research resulted in the development of a plasma textile that can be used for producing new types of self-decontaminating garments, fabrics, and filter materials, capable of activating a plasma sheath that would filter, capture, and destroy any bacteriological agent deposited on its surface. This new material relies on the unique antimicrobial and catalytic properties of cold (room temperature) plasma that is benign to people and does not cause thermal damage to many polymer textiles, such as Nomex and polypropylene. The uniqueness of cold plasma as a disinfecting agent lies in the inability of bacteria to develop resistance to plasma exposure, as they can for antibiotics. Plasma textiles could thus be utilized for microbial destruction in active antimicrobial filters (for continuous decontamination and disinfection of large amounts of air) as well as in self-decontaminating surfaces and antibacterial barriers (for example, for creating local antiseptic or sterile environments around wounds and burns).

  15. Surface properties of activated carbon treated by cold plasma heating

    Energy Technology Data Exchange (ETDEWEB)

    Norikazu, Kurano [Shigematsu works Co. Ltd., 267 Yashita, Iwatsuki 3390046 (Japan); Yamada, Hiroshi [Shigematsu works Co. Ltd., 267 Yashita, Iwatsuki 3390046 (Japan); Yajima, Tatsuhiko [Faculty of Engineering, Saitama Institute of Technology, 1690 Fusoiji, Okabe 3690293 (Japan); Sugiyama, Kazuo [Faculty of Engineering, Saitama University, 255 Shimo-okubo, Sakura-Ku, Saitama 3388570 (Japan)]. E-mail: sugi@apc.saitama-u.ac.jp

    2007-03-12

    To modify the surface properties of activated carbon powders, we have applied the cold plasma treatment method. The cold plasma was used to be generated in the evacuated reactor vessel by 2.45 GHz microwave irradiation. In this paper, changes of surface properties such as distribution of acidic functional groups and roughness morphology were examined. By the cold plasma treatment, activated carbons with large specific surface area of ca. 2000 m{sup 2}/g or more could be prepared in a minute. The amount of every gaseous organic compound adsorbed on the unit gram of treated activated carbons was more increased that on the unit gram of untreated carbons. Especially, the adsorbed amount of carbon disulfide was remarkably increased even if it was compared by the amount per unit surface area. These results suggest that the surface property of the sample was modified by the plasma treatment. It became apparent by observing SEM photographs that dust and impure particles in macropores of activated carbons were far more reduced by the plasma treatment than by the conventional heating in an electric furnace under vacuum. In addition, a bubble-like surface morphology of the sample was observed by AEM measurement. The amount of acidic functional groups at the surface was determined by using the Boehm's titration method. Consequently, the increase of lactone groups and the decrease of carboxyl groups were also observed.

  16. Modulation of Erythrocyte Plasma Membrane Redox System Activity by Curcumin

    Directory of Open Access Journals (Sweden)

    Prabhakar Singh

    2016-01-01

    Full Text Available Plasma membrane redox system (PMRS is an electron transport chain system ubiquitously present throughout all cell types. It transfers electron from intracellular substrates to extracellular acceptors for regulation of redox status. Curcumin, isolated from Curcuma longa, has modulatory effects on cellular physiology due to its membrane interaction ability and antioxidant potential. The present study investigates the effect of curcumin on PMRS activity of erythrocytes isolated from Wistar rats in vitro and in vivo and validated through an in silico docking simulation study using Molegro Virtual Docker (MVD. Effects of curcumin were also evaluated on level of glutathione (GSH and the oxidant potential of plasma measured in terms of plasma ferric equivalent oxidative potentials (PFEOP. Results show that curcumin significantly (p<0.01 downregulated the PMRS activity in a dose-dependent manner. Molecular docking results suggest that curcumin interacts with amino acids at the active site cavity of cytochrome b5 reductase, a key constituent of PMRS. Curcumin also increased the GSH level in erythrocytes and plasma while simultaneously decreasing the oxidant potential (PFEOP of plasma. Altered PMRS activity and redox status are associated with the pathophysiology of several health complications including aging and diabetes; hence, the above finding may explain part of the role of curcumin in health beneficial effects.

  17. Plasma Treated Active Carbon for Capacitive Deionization of Saline Water

    Directory of Open Access Journals (Sweden)

    Aiping Zeng

    2017-01-01

    Full Text Available The plasma treatment on commercial active carbon (AC was carried out in a capacitively coupled plasma system using Ar + 10% O2 at pressure of 4.0 Torr. The RF plasma power ranged from 50 W to 100 W and the processing time was 10 min. The carbon film electrode was fabricated by electrophoretic deposition. Micro-Raman spectroscopy revealed the highly increased disorder of sp2 C lattice for the AC treated at 75 W. An electrosorption capacity of 6.15 mg/g was recorded for the carbon treated at 75 W in a 0.1 mM NaCl solution when 1.5 V was applied for 5 hours, while the capacity of the untreated AC was 1.01 mg/g. The plasma treatment led to 5.09 times increase in the absorption capacity. The jump of electrosorption capacity by plasma treatment was consistent with the Raman spectra and electrochemical double layer capacitance. This work demonstrated that plasma treatment was a potentially efficient approach to activating biochar to serve as electrode material for capacitive deionization (CDI.

  18. Improvement of Raw-Starch Digestibility of Amylase from Aspergillus awamori by Gamma Radiation Mutation for Alcohol Production

    International Nuclear Information System (INIS)

    Kanlayakrit, Werasit; Piadang, Nattayana; Maweang, Metinee

    2006-09-01

    Aspergillus awamori was induced to mutation by gamma ray to improve raw-starch digestibility of amylase enzyme. Twenty fungal colonies were selected base on and amylase and glucoamylase activities including raw starch digestibility. The result showed that isolate A a(i)-2(16) was the best isolate for raw-starch digestion (65.64 %). It produced extracellular amylase enzyme which showed highest raw-starch digestibility more than wild type about 2 folds. Based on enzymes from solid culture showed activities higher from liquid medium. Therefore solid culture is suitable for fungal enzyme production.

  19. Plasma edge physics in an actively cooled tokamak

    International Nuclear Information System (INIS)

    Gunn, J.P.; Adamek, A.; Boucher, C.

    2005-01-01

    Tore Supra is a large tokamak with a plasma of circular cross section (major radius 2.4 m and minor radius 0.72 m) lying on a toroidal limiter. Tore Supra's main mission is the development of technology to inject up to 25 MW of microwave heating power and extract it continuously for up to 1000 s in steady state without uncontrolled overheating of, or outgassing from, plasma-facing components. The entire first wall of the tokamak is actively cooled by a high pressure water loop and special carbon fiber composite materials have been designed to handle power fluxes up to 10 MW/m 2 . The edge plasma on open magnetic flux surfaces that intersect solid objects plays an important role in the overall behaviour of the plasma. The transport of sputtered impurity ions and the fueling of the core plasma are largely governed by edge plasma density, temperature, and flow profiles. Measurements of these quantities are becoming more reliable and frequent in many tokamaks, and it has become clear that we do not understand them very well. Classical two-dimensional fluid modelling fails to reproduce many aspects of the experimental observations such as the significant thickness of the edge plasma, and the near-sonic flows that occur where none should be expected. It is suspected that plasma turbulence is responsible for these anomalies. In the Tore Supra tokamak, various kinds of Langmuir probes are used to characterize the edge plasma. We will present original measurements that demonstrate the universality of many phenomena that have been observed in X-point divertor tokamaks, especially concerning the ion flows. As in the JET tokamak, surprisingly large values of parallel Mach number are measured midway between the two strike zones, where one would expect to find nearly stagnant plasma if the particle source were poloidally uniform. We will present results of a novel experiment that provides evidence for a poloidally localized particle and energy source on the outboard midplane of

  20. Production, Purification, and Characterization of Thermostable α-Amylase Produced by Bacillus licheniformis Isolate AI20

    Directory of Open Access Journals (Sweden)

    Yasser R. Abdel-Fattah

    2013-01-01

    Full Text Available An optimization strategy, based on statistical experimental design, is employed to enhance the production of thermostable α-amylase by a thermotolerant B. licheniformis AI20 isolate. Using one variant at time (OVAT method, starch, yeast extract, and CaCl2 were observed to influence the enzyme production significantly. Thereafter, the response surface methodology (RSM was adopted to acquire the best process conditions among the selected variables, where a three-level Box-Behnken design was employed to create a polynomial quadratic model correlating the relationship between the three variables and α-amylase activity. The optimal combination of the major constituents of media for α-amylase production was 1.0% starch, 0.75% yeast extract, and 0.02% CaCl2. The predicted optimum α-amylase activity was 384 U/mL/min, which is two folds more than the basal medium conditions. The produced α-amylase was purified through various chromatographic techniques. The estimated enzyme molecular mass was 55 kDa and the α-amylase had an optimal temperature and pH of 60–80°C and 6–7.5, respectively. Values of Vmax and Km for the purified enzyme were 454 mU/mg and 0.709 mg/mL. The α-amylase enzyme showed great stability against different solvents. Additionally, the enzyme activity was slightly inhibited by detergents, sodium dodecyl sulphate (SDS, or chelating agents such as EDTA and EGTA. On the other hand, great enzyme stability against different divalent metal ions was observed at 0.1 mM concentration, but 10 mM of Cu2+ or Zn2+ reduced the enzyme activity by 25 and 55%, respectively.

  1. Transglycosylation by barley α-amylase 1

    DEFF Research Database (Denmark)

    Mótyán, János A.; Fazekas, Erika; Mori, Haruhide

    2011-01-01

    The transglycosylation activity of barley α-amylase 1 (AMY1) and active site AMY1 subsite mutant enzymes was investigated. We report here the transferase ability of the V47A, V47F, V47D and S48Y single mutants and V47K/S48G and V47G/S48D double mutant AMY1 enzymes in which the replaced amino acids...... play important role in substrate binding at subsites at −3 through −5. Although mutation increases the transglycosylation activity of enzymes, in the presence of acceptors the difference between wild type and mutants is not so significant. Oligomer transfer reactions of AMY1 wild type and its mutants...... as donor. 4-Methylumbelliferyl-α-d-maltoside, -maltotrioside, -maltotetraoside and -maltopentaoside have been synthesized. Products were identified by MALDI-TOF MS. 1H and 13C NMR analyses showed that AMY1 V47F preserved the stereo- and regioselectivity. The produced MU-α-d-MOSs of degree of polymerization...

  2. Contribution of Musa paradisiaca in the inhibition of α-amylase, α-glucosidase and Angiotensin-I converting enzyme in streptozotocin induced rats.

    Science.gov (United States)

    Shodehinde, Sidiqat A; Ademiluyi, Adedayo O; Oboh, Ganiyu; Akindahunsi, Afolabi A

    2015-07-15

    Unripe plantain based-diets are part of folklore remedy for the management of diabetes in tropical Africa; however, with the dearth of information on the rationale behind this practice; this study therefore, sought to investigate the antihyperglycemic effect of traditional unripe plantain products (Amala and Booli) in high fat fed/low dose streptozotocin-induced diabetic rats and to provide a possible rationale for their antidiabetic properties. Diabetes was induced experimentally by high fat fed/low dose streptozotocin-diabetic rats (25mg/kg body wt.) and the diabetic rats were fed diets supplemented with 20-40% Amala and Booli for 14 days. The effect of the diets on the blood glucose level, pancreatic α-amylase, intestinal α-glucosidase and Angiotensin-I converting enzyme (ACE) activities and plasma antioxidant status as well as amylose/amylopectin content of the unripe plantain products were determined. A marked increase in the blood glucose, α-amylase, α-glucosidase and ACE activities with a corresponding decrease in plasma antioxidant status was recorded in diabetic rats. However, these indices were significantly (P < 0.05) reversed after unripe plantain product supplemented diet treatments for 14 days. Also, the amylose/amylopectin ratio of the products is 1:3. This study revealed that unripe plantain products exert antihyperglycemic effects which could be attributed to the inhibition of α-amylase and α-glucosidase activities by their constituent phytochemicals as well as their amylose/amylopectin contents in the diabetic rats, hence, providing the possible rationale behind their antidiabetic properties. Copyright © 2015 Elsevier Inc. All rights reserved.

  3. Production and characterization of amylases from Zea mays malt

    Directory of Open Access Journals (Sweden)

    Joana Paula Menezes Biazus

    2009-08-01

    Full Text Available In this work the α and β-amylase enzymes were obtained from maize (Zea mays malt and were biochemistry characterized. A germination study to obtain the maize malt with good amylase activity was made. The maize seeds were germinated in laboratory and the enzymatic activity was measured daily. Activity dependence to germination time were fitted to an exponential model (A=A0eµt, which showed that the behaviour of enzymatic activity in the germination process was similar to the growth of the microorganism. Its model could be applied to describe the mechanism of α-amylase production for each maize varieties and others cereals. Maize malt characterization showed that α and β-amylase had optimal pH between 4-6.5, optimal temperature 50 and 90ºC, and molecular weight of 67.4 and 47.5kDa, respectively. This work contributed with the advances in biotechnology generating of conditions for application of a new and of low price amylases source.Neste trabalho as enzimas α e β-amilases foram obtidas de malte de milho e depois foram caracterizadas bioquimicamente. Um estudo da germinação foi feito para obtenção do malte com boa atividade amilásica. A germinação ocorreu em escala laboratorial e a atividade enzimática foi medida diariamente. Um modelo exponencial do tipo A=A0eµt foi ajustado a dependência do tempo de germinação com a atividade, mostrando que o comportamento da atividade enzimática no processo de germinação é semelhante ao crescimento de microorganismos. Este modelo pode ser aplicado para descrever o mecanismo de produção da α-amilase para cada variedade de milho e de outros cereais. A caracterização do malte de milho mostrou que as α e β-amilase têm pH ótimo entre 4,0-6,5, temperatura ótima de 50 e 90ºC, e massa molar de 67,4 e 47,5 kDa, respectivamente. Este trabalho contribuiu com os avanços da biotecnologia gerando condições de emprego de uma nova e barata fonte de amilases.

  4. Conditions of activation of yeast plasma membrane ATPase.

    Science.gov (United States)

    Sychrová, H; Kotyk, A

    1985-04-08

    The in vivo activation of the H+-ATPase of baker's yeast plasma membrane found by Serrano in 1983 was demonstrated with D-glucose aerobically and anaerobically (as well as in a respiration-deficient mutant) and, after suitable induction, with maltose, trehalose, and galactose. The activated but not the control ATPase was sensitive to oligomycin. No activation was possible in a cell-free extract with added glucose. The ATPase was not activated in yeast protoplasts which may account for the absence of glucose-stimulated secondary active transports in these wall-less cells and provide support for a microscopic coupling between ATPase activity and these transports in yeast cells.

  5. Characterizing electrostatic turbulence in tokamak plasmas with high MHD activity

    Energy Technology Data Exchange (ETDEWEB)

    Guimaraes-Filho, Z O; Santos Lima, G Z dos; Caldas, I L; Nascimento, I C; Kuznetsov, Yu K [Instituto de Fisica, Universidade de Sao Paulo, Caixa Postal 66316, 05315-970, Sao Paulo, SP (Brazil); Viana, R L, E-mail: viana@fisica.ufpr.b [Departamento de Fisica, Universidade Federal do Parana, Caixa Postal 19044, 81531-990, Curitiba, PR (Brazil)

    2010-09-01

    One of the challenges in obtaining long lasting magnetic confinement of fusion plasmas in tokamaks is to control electrostatic turbulence near the vessel wall. A necessary step towards achieving this goal is to characterize the turbulence level and so as to quantify its effect on the transport of energy and particles of the plasma. In this paper we present experimental results on the characterization of electrostatic turbulence in Tokamak Chauffage Alfven Bresilien (TCABR), operating in the Institute of Physics of University of Sao Paulo, Brazil. In particular, we investigate the effect of certain magnetic field fluctuations, due to magnetohydrodynamical (MHD) instabilities activity, on the spectral properties of electrostatic turbulence at plasma edge. In some TCABR discharges we observe that this MHD activity may increase spontaneously, following changes in the edge safety factor, or after changes in the radial electric field achieved by electrode biasing. During the high MHD activity, the magnetic oscillations and the plasma edge electrostatic turbulence present several common linear spectral features with a noticeable dominant peak in the same frequency. In this article, dynamical analyses were applied to find other alterations on turbulence characteristics due to the MHD activity and turbulence enhancement. A recurrence quantification analysis shows that the turbulence determinism radial profile is substantially changed, becoming more radially uniform, during the high MHD activity. Moreover, the bicoherence spectra of these two kinds of fluctuations are similar and present high bicoherence levels associated with the MHD frequency. In contrast with the bicoherence spectral changes, that are radially localized at the plasma edge, the turbulence recurrence is broadly altered at the plasma edge and the scrape-off layer.

  6. Improving the Yield of Glucoamylase and α-amylase in Solid-state Co-culture

    Directory of Open Access Journals (Sweden)

    Takefuji Haruka

    2016-01-01

    Full Text Available Steamed rice inoculated with Aspergillus oryzae, called Koji, is an essential ingredient for making amazake or brewing sake. However, A. oryzae usually offers low enzyme yield, especially in case of glucoamylase. Co-culture of Aspergillus and Rhizopus strains in Koji increased glucoamylase and α-amylase activities. The ratio of initial spore counts of A. oryzae and R. oryzae influenced the activity of amylolytic enzymes. When this ratio was 1:1, α-amylase showed maximum activity (573 U/g-substrate, 95 h, and when this ratio was 200:1, glucoamylase showed maximum activity (180 U/g-substrate, 95 h.

  7. Optimization, Purification, and Starch Stain Wash Application of Two New α-Amylases Extracted from Leaves and Stems of Pergularia tomentosa

    Directory of Open Access Journals (Sweden)

    Imen Lahmar

    2017-01-01

    Full Text Available A continuous research is attempted to fulfil the highest industrial demands of natural amylases presenting special properties. New α-amylases extracted from stems and leaves of Pergularia tomentosa, which is widespread and growing spontaneously in Tunisia, were studied by the means of their activities optimization and purification. Some similarities were recorded for the two identified enzymes: (i the highest amylase activity showed a promoted thermal stability at 50°C; (ii the starch substrate at 1% enhanced the enzyme activity; (iii the two α-amylases seem to be calcium-independent; (iv Zn2+, Cu2+, and Ag2+ were considered as important inhibitors of the enzyme activity. Following the increased gradient of elution on Mono Q-Sepharose column, an increase in the specific activity of 11.82-fold and 10.92-fold was recorded, respectively, for leaves and stems with the presence of different peaks on the purification profiles. Pergularia amylases activities were stable and compatible with the tested commercial detergents. The combination of plant amylase and detergent allowed us to enhance the wash performance with an increase of 35.24 and 42.56%, respectively, for stems and leaves amylases. Characterized amylases were reported to have a promoted potential for their implication notably in detergent industry as well as biotechnological sector.

  8. Improved α-Amylase Production by Dephosphorylation Mutation of CreD, an Arrestin-Like Protein Required for Glucose-Induced Endocytosis of Maltose Permease and Carbon Catabolite Derepression in Aspergillus oryzae.

    Science.gov (United States)

    Tanaka, Mizuki; Hiramoto, Tetsuya; Tada, Hinako; Shintani, Takahiro; Gomi, Katsuya

    2017-07-01

    Aspergillus oryzae produces copious amount of amylolytic enzymes, and MalP, a major maltose permease, is required for the expression of amylase-encoding genes. The expression of these genes is strongly repressed by carbon catabolite repression (CCR) in the presence of glucose. MalP is transported from the plasma membrane to the vacuole by endocytosis, which requires the homolog of E6-AP carboxyl terminus ubiquitin ligase HulA, an ortholog of yeast Rsp5. In yeast, arrestin-like proteins mediate endocytosis as adaptors of Rsp5 and transporters. In the present study, we examined the involvement of CreD, an arrestin-like protein, in glucose-induced MalP endocytosis and CCR of amylase-encoding genes. Deletion of creD inhibited the glucose-induced endocytosis of MalP, and CreD showed physical interaction with HulA. Phosphorylation of CreD was detected by Western blotting, and two serine residues were determined as the putative phosphorylation sites. However, the phosphorylation state of the serine residues did not regulate MalP endocytosis and its interaction with HulA. Although α-amylase production was significantly repressed by creD deletion, both phosphorylation and dephosphorylation mimics of CreD had a negligible effect on α-amylase activity. Interestingly, dephosphorylation of CreD was required for CCR relief of amylase genes that was triggered by disruption of the deubiquitinating enzyme-encoding gene creB The α-amylase activity of the creB mutant was 1.6-fold higher than that of the wild type, and the dephosphorylation mimic of CreD further improved the α-amylase activity by 2.6-fold. These results indicate that a combination of the dephosphorylation mutation of CreD and creB disruption increased the production of amylolytic enzymes in A. oryzae IMPORTANCE In eukaryotes, glucose induces carbon catabolite repression (CCR) and proteolytic degradation of plasma membrane transporters via endocytosis. Glucose-induced endocytosis of transporters is mediated by

  9. α-Amylase in Vaginal Fluid: Association With Conditions Favorable to Dominance of Lactobacillus.

    Science.gov (United States)

    Nasioudis, Dimitrios; Beghini, Joziani; Bongiovanni, Ann Marie; Giraldo, Paulo C; Linhares, Iara M; Witkin, Steven S

    2015-11-01

    Vaginal glycogen is degraded by host α-amylase and then converted to lactic acid by Lactobacilli. This maintains the vaginal pH at ≤4.5 and prevents growth of other bacteria. Therefore, host α-amylase activity may promote dominance of Lactobacilli. We evaluated whether the α-amylase level in vaginal fluid is altered in women with bacterial vaginosis (BV) and vulvovaginal candidiasis (VVC) and whether its concentration was associated with levels of lactic acid isomers and host mediators. Vaginal fluid was obtained from 43 women with BV, 50 women with VVC, and 62 women with no vulvovaginal disorders. Vaginal fluid concentrations of α-amylase, secretory leukocyte protease inhibitor (SLPI), hyaluronan, hyaluronidase-1, β-defensin, and elafin were measured by enzyme-linked immunosorbent assay (ELISA). Vaginal concentrations of neutrophil gelatinase-associated lipocalin (NGAL), matrix metalloproteinase (MMP) 8, and d- and l-lactic acid levels in these patients were previously reported. The median vaginal fluid α-amylase level was 1.83 mU/mL in control women, 1.45 mU/mL in women with VVC, and 1.07 mU/mL in women with BV. Vaginal levels of α-amylase were correlated with d-lactic acid (P = .003) but not with l-lactic acid (P > .05) and with SLPI (P vaginal lumen by hyaluronidase-1 and MMP-8 may increase glycogen availability and promote α-amylase activity. The subsequent enhanced availability of glycogen breakdown products would favor proliferation of Lactobacilli, the primary producers of d-lactic acid in the vagina. Concomitant production of NGAL and SLPI would retard growth of BV-related bacteria. © The Author(s) 2015.

  10. Evolution of active region loop plasma

    International Nuclear Information System (INIS)

    Krall, K.R.; Antiochos, S.K.

    1980-01-01

    We investigate numerically the adjustment of coronal active-region loops to changes in their heating rate. The one-dimensional hydrodynamic equations are solved subject to boundary conditions in which heat flux-induced mass exchange between coronal and chromospheric components is allowed. The calculated evolution of physical parameters suggests that (1) mass supplied during chromospheric evaporation is much more effective in moderating coronal temperature excursions than when downward heat flux if dissipated by a static chromosphere, and (2) the method by which rhe chromosphere responds to changing coronal conditions can significantly influence coronal readjustment time scales. Observations are cited which illustrate the range of possible fluctuations in the heating rates

  11. Fission and activation of uranium by fashion-plasma neutrons

    International Nuclear Information System (INIS)

    Lee, J.H.; Hochl, F.; McFarland, D.R.

    1978-01-01

    Disks of enriched and depleted uranium were irradiated by neutrons from the D-D fusions in a dense plasma-focus. A fission yield of 10 6 fissions-cm -3 in U 235 per pulse was determined with Ge(Li) gamme-ray spectrometry. Activation of U 238 caused increased beta activity after the plasma-neutron irradiation but alpha-particle spectrometry showed Pu 239 production was negligible. In addition, with a disk of lithium in the apparatus, 13.3 MeV neutrons from 7 Li(d,n) 8 Be was observed with a 80-m time-of-flight neutron detector. Dense plasma focuses are now operated not only in a single coaxial gun, but also in improved geometries, such as the hypocycloidal pinch and the staged plasma focus, from which a multiple plasma-focus array suitable for experimental verification of, and eventuel development into a fusion-fission hybrid reactor could be produced. (orig.) [de

  12. Mutational analysis of target enzyme recognition of the beta-trefoil fold barley alpha-amylase/subtilisin inhibitor

    DEFF Research Database (Denmark)

    Bønsager, Birgit Christine; Nielsen, Per K.; Abou Hachem, Maher

    2005-01-01

    The barley alpha-amylase/subtilisin inhibitor ( BASI) inhibits alpha-amylase 2 (AMY2) with subnanomolar affinity. The contribution of selected side chains of BASI to this high affinity is discerned in this study, and binding to other targets is investigated. Seven BASI residues along the AMY2-BASI...... interface and four residues in the putative protease-binding loop on the opposite side of the inhibitor were mutated. A total of 15 variants were compared with the wild type by monitoring the alpha-amylase and protease inhibitory activities using Blue Starch and azoalbumin, respectively, and the kinetics...

  13. Novel Mechanism of Plasma Prekallikrein (PK) Activation by Vascular Smooth Muscle Cells: Evidence of the presence of PK Activator

    OpenAIRE

    Keum, Joo-Seob; Jaffa, Miran A; Luttrell, Louis M; Jaffa, Ayad A.

    2014-01-01

    The contribution of plasma prekallikrein (PK) to vascular remodeling is becoming increasingly recognized. Plasma PK is activated when the zymogen PK is digested to an active enzyme by activated factor XII (FXII). Here, we present our findings that vascular smooth muscle cells (VSMC) activate plasma PK in the absence of FXII. Extracted plasma membrane and cytosolic fractions of VSMCs activate PK, but the rate of PK activation was greater by the membrane fraction. FXII neutralizing antibody did...

  14. Pancreatic α-Amylase Controls Glucose Assimilation by Duodenal Retrieval through N-Glycan-specific Binding, Endocytosis, and Degradation*

    Science.gov (United States)

    Date, Kimie; Satoh, Ayano; Iida, Kaoruko; Ogawa, Haruko

    2015-01-01

    α-Amylase, a major pancreatic protein and starch hydrolase, is essential for energy acquisition. Mammalian pancreatic α-amylase binds specifically to glycoprotein N-glycans in the brush-border membrane to activate starch digestion, whereas it significantly inhibits glucose uptake by Na+/glucose cotransporter 1 (SGLT1) at high concentrations (Asanuma-Date, K., Hirano, Y., Le, N., Sano, K., Kawasaki, N., Hashii, N., Hiruta, Y., Nakayama, K., Umemura, M., Ishikawa, K., Sakagami, H., and Ogawa, H. (2012) Functional regulation of sugar assimilation by N-glycan-specific interaction of pancreatic α-amylase with glycoproteins of duodenal brush border membrane. J. Biol. Chem. 287, 23104–23118). However, how the inhibition is stopped was unknown. Here, we show a new mechanism for the regulation of intestinal glucose absorption. Immunohistochemistry revealed that α-amylase in the duodena of non-fasted, but not fasted, pigs was internalized from the pancreatic fluid and immunostained. We demonstrated that after N-glycan binding, pancreatic α-amylase underwent internalization into lysosomes in a process that was inhibited by α-mannoside. The internalized α-amylase was degraded, showing low enzymatic activity and molecular weight at the basolateral membrane. In a human intestinal Caco-2 cell line, Alexa Fluor 488-labeled pancreatic α-amylase bound to the cytomembrane was transported to lysosomes through the endocytic pathway and then disappeared, suggesting degradation. Our findings indicate that N-glycan recognition by α-amylase protects enterocytes against a sudden increase in glucose concentration and restores glucose uptake by gradual internalization, which homeostatically controls the postprandial blood glucose level. The internalization of α-amylase may also enhance the supply of amino acids required for the high turnover of small intestine epithelial cells. This study provides novel and significant insights into the control of blood sugar during the absorption

  15. Improving the Yield of Glucoamylase and α-amylase in Solid-state Co-culture

    OpenAIRE

    Takefuji Haruka; Ninomiya Junko; Morita Hiroshi

    2016-01-01

    Steamed rice inoculated with Aspergillus oryzae, called Koji, is an essential ingredient for making amazake or brewing sake. However, A. oryzae usually offers low enzyme yield, especially in case of glucoamylase. Co-culture of Aspergillus and Rhizopus strains in Koji increased glucoamylase and α-amylase activities. The ratio of initial spore counts of A. oryzae and R. oryzae influenced the activity of amylolytic enzymes. When this ratio was 1:1, α-amylase showed maximum activity (573 U/g-subs...

  16. Contribution to the Chemistry of Plasma-Activated Water

    Science.gov (United States)

    Julák, J.; Hujacová, A.; Scholtz, V.; Khun, J.; Holada, K.

    2018-01-01

    Plasma-activated water (PAW) was prepared by exposure to nonthermal plasma produced by a positive dc corona discharge in a transient spark regime. The activation of water was performed in atmosphere of various surrounding gases (air, nitrogen, carbon dioxide, and argon). This PAW retains its biological activity, measured on the mouse neuroblastoma cells culture, even after storage for more than one year. The highest hydrogen peroxide content was found for PAWs prepared in the atmospheres of argon or carbon dioxide, whereas the PAWs prepared in air and nitrogen exhibited lower hydrogen peroxide content. The acidity of PAWs mediated by nitric and nitrous acid formation displayed an opposite trend. It is concluded that the long-lasting biological effect of PAW is mediated by hydrogen peroxide in acid milieu only, whereas other possible active components decompose rapidly.

  17. Geomagnetic activity effects on plasma sheet energy conversion

    Directory of Open Access Journals (Sweden)

    M. Hamrin

    2010-10-01

    Full Text Available In this article we use three years (2001, 2002, and 2004 of Cluster plasma sheet data to investigate what happens to localized energy conversion regions (ECRs in the plasma sheet during times of high magnetospheric activity. By examining variations in the power density, E·J, where E is the electric field and J is the current density obtained by Cluster, we have studied the influence on Concentrated Load Regions (CLRs and Concentrated Generator Regions (CGRs from variations in the geomagnetic disturbance level as expressed by the Kp, the AE, and the Dst indices. We find that the ECR occurrence frequency increases during higher magnetospheric activities, and that the ECRs become stronger. This is true both for CLRs and for CGRs, and the localized energy conversion therefore concerns energy conversion in both directions between the particles and the fields in the plasma sheet. A higher geomagnetic activity hence increases the general level of energy conversion in the plasma sheet. Moreover, we have shown that CLRs live longer during magnetically disturbed times, hence converting more electromagnetic energy. The CGR lifetime, on the other hand, seems to be unaffected by the geomagnetic activity level. The evidence for increased energy conversion during geomagnetically disturbed times is most clear for Kp and for AE, but there are also some indications that energy conversion increases during large negative Dst. This is consistent with the plasma sheet magnetically mapping to the auroral zone, and therefore being more tightly coupled to auroral activities and variations in the AE and Kp indices, than to variations in the ring current region as described by the Dst index.

  18. Inhibition of pancreatic lipase and amylase by extracts of different spices and plants.

    Science.gov (United States)

    Sellami, Mohamed; Louati, Hanen; Kamoun, Jannet; Kchaou, Ali; Damak, Mohamed; Gargouri, Youssef

    2017-05-01

    The aim of this study is to search new anti-obesity and anti-diabetic agents from plant and spices crude extracts as alternative to synthetic drugs. The inhibitory effect of 72 extracts was evaluated, in vitro, on lipase and amylase activities. Aqueous extracts of cinnamon and black tea exhibited an appreciable inhibitory effect on pancreatic amylase with IC 50 values of 18 and 87 μg, respectively. Aqueous extracts of cinnamon and mint showed strong inhibitory effects against pancreatic lipase with IC 50 of 45 and 62 μg, respectively. The presence of bile salts and colipase or an excess of interface failed to restore the lipase activity. Therefore, the inhibition of pancreatic lipase, by extracts of spices and plants, belongs to an irreversible inhibition. Crude extract of cinnamon showed the strongest anti-lipase and anti-amylase activities which offer a prospective therapeutic approach for the management of diabetes and obesity.

  19. A radioreceptor assay for measurement of plasma glucocorticoid binding activity

    International Nuclear Information System (INIS)

    Fan Jie

    1990-01-01

    A radioreceptor assay (RRA) for plasma glucocorticoid binding activity (GCBA) has been developed using glucocorticoid receptor in rat thymocytes. Unlike other assays for natural and certain synthetic corticosteroids, RRA measures the GCBA of all natural and synthetic GC in plasma. The range of standard curve was 0 ∼ 1.00 mg/L. The sensitivity was 0.01 mg/l. The recovery rate was 92.1%, and the intra and inter assay CV was 0.7% (n = 3) and 4.4% (n = 3) respectively. The level of corticosterone in 9 rat plasma samples was determined by RRA and CBG-isotope binding assay. There was a general correlation over a wide range between the values determined by the two assays (r = 0.95; P < 0.001). The measuring condition was described in detail

  20. Improved production, purification and some properties of α-amylase ...

    African Journals Online (AJOL)

    user

    2012-10-04

    Oct 4, 2012 ... nology industries with many potential applications in food, fermentation, textile and .... Large scale production of α-amylase enzyme in a laboratory fermentor ..... induction on amylase production and low cost (Li et al.,. 2010).

  1. Genetic analyses using GGE model and a mixed linear model approach, and stability analyses using AMMI bi-plot for late-maturity alpha-amylase activity in bread wheat genotypes.

    Science.gov (United States)

    Rasul, Golam; Glover, Karl D; Krishnan, Padmanaban G; Wu, Jixiang; Berzonsky, William A; Fofana, Bourlaye

    2017-06-01

    Low falling number and discounting grain when it is downgraded in class are the consequences of excessive late-maturity α-amylase activity (LMAA) in bread wheat (Triticum aestivum L.). Grain expressing high LMAA produces poorer quality bread products. To effectively breed for low LMAA, it is necessary to understand what genes control it and how they are expressed, particularly when genotypes are grown in different environments. In this study, an International Collection (IC) of 18 spring wheat genotypes and another set of 15 spring wheat cultivars adapted to South Dakota (SD), USA were assessed to characterize the genetic component of LMAA over 5 and 13 environments, respectively. The data were analysed using a GGE model with a mixed linear model approach and stability analysis was presented using an AMMI bi-plot on R software. All estimated variance components and their proportions to the total phenotypic variance were highly significant for both sets of genotypes, which were validated by the AMMI model analysis. Broad-sense heritability for LMAA was higher in SD adapted cultivars (53%) compared to that in IC (49%). Significant genetic effects and stability analyses showed some genotypes, e.g. 'Lancer', 'Chester' and 'LoSprout' from IC, and 'Alsen', 'Traverse' and 'Forefront' from SD cultivars could be used as parents to develop new cultivars expressing low levels of LMAA. Stability analysis using an AMMI bi-plot revealed that 'Chester', 'Lancer' and 'Advance' were the most stable across environments, while in contrast, 'Kinsman', 'Lerma52' and 'Traverse' exhibited the lowest stability for LMAA across environments.

  2. Expression of alpha-amylase in Bacillus licheniformis.

    OpenAIRE

    Rothstein, D M; Devlin, P E; Cate, R L

    1986-01-01

    In Bacillus licheniformis, alpha-amylase production varied more than 100-fold depending on the presence or absence of a catabolite-repressing carbon source in the growth medium. alpha-Amylase was produced during the growth phase and not at the onset of the stationary phase. Induction of alpha-amylase correlated with synthesis of mRNA initiating at the promoter of the alpha-amylase gene.

  3. Aminotransferases and Leucine Aminopeptidase Activity in Blood Plasma of Chickens

    International Nuclear Information System (INIS)

    Kraljevic, P.; Stojevic, Z.; Milinkovic-Tur, S.; Simpraga, M.; Miljanic, S.

    1998-01-01

    It has been reported that irradiation of mammals by gama-rays cause increase of some enzyme activity in their blood plasma (Miller and Gates 1949; Milch and Albaum 1959; Hughes 1958; Miholjcic et al. 1979). In our previous papers (Kraljevic et al., 1982; Kraljevic and Emanovic 1993) it has been shown that activities of some enzymes in the blood plasma of chickens after an intramuscular injection of radioactive isotope 32 P. In this paper an attempt has been made to investigate the influence of gamma-ray irradiation of the whole body of chickens upon activity of some enzymes in their blood plasma. We also wanted to investigate whether the activity of aspartate-aminotransferase (AST), alanine aminotransferase (ALT) and leucine-aminopeptidase (LAP) may serve as an additional test for functional liver damage in chickens caused by gamma-ray. Fifty day old hybrid male chickens of heavy Jata breeds were irradiated by gamma-ray in the dose of 7,23±0,95 Gy. Blood samples were taken from the wing vein on days 1, 3, 5, 7, 9 and 15 after irradiation. Activity of AST, ALT, and LAP in the blood plasma were determined spectrophotometrically using Boehringer Mannheim GmbH optimized kits. At the end of the experiment all birds were sacrificed and, as well as died birds were photomorphologically and histologically investigated. The obtained results showed decrease of activity of all three enzymes during the whole period of investigation, but significant decrease showed only AST and LAP. It seems that both enzymes may serve as additional test for functional liver damage in chickens by external gamma-rays. (author)

  4. Amylase production under solid state fermentation by a bacterial ...

    African Journals Online (AJOL)

    This study was concerned with the screening of a suitable isolate and optimization of cultural conditions for the biosynthesis of thermostable amylase under solid state fermentation (SSF). Twenty seven isolates were screened for amylase production out of which one isolate designated as W74 showed maximal amylase ...

  5. High Temperature-Induced Expression of Rice α-Amylases in Developing Endosperm Produces Chalky Grains

    Directory of Open Access Journals (Sweden)

    Masaru Nakata

    2017-12-01

    Full Text Available Global warming impairs grain filling in rice and reduces starch accumulation in the endosperm, leading to chalky-appearing grains, which damages their market value. We found previously that high temperature-induced expression of starch-lytic α-amylases during ripening is crucial for grain chalkiness. Because the rice genome carries at least eight functional α-amylase genes, identification of the α-amylase(s that contribute most strongly to the production of chalky grains could accelerate efficient breeding. To identify α-amylase genes responsible for the production of chalky grains, we characterized the histological expression pattern of eight α-amylase genes and the influences of their overexpression on grain appearance and carbohydrate components through a series of experiments with transgenic rice plants. The promoter activity of most α-amylase genes was elevated to various extents at high temperature. Among them, the expression of Amy1A and Amy3C was induced in the internal, especially basal to dorsal, region of developing endosperm, whereas that of Amy3D was confined near the ventral aleurone. These regions coincided with the site of occurrence of chalkiness, which was in clear contrast to conventionally known expression patterns of the enzyme in the scutellum and aleurone during seed germination. Furthermore, overexpression of α-amylase genes, except for Amy3E, in developing endosperm produced various degrees of chalky grains without heat exposure, whereas that of Amy3E yielded normal translucent grains, as was the case in the vector control, even though Amy3E-overexpressing grains contained enhanced α-amylase activities. The weight of the chalky grains was decreased due to reduced amounts of starch, and microscopic observation of the chalky part of these grains revealed that their endosperm consisted of loosely packed round starch granules that had numerous pits on their surface, confirming the hydrolysis of the starch reserve by

  6. Immunoradiometric quantitation of tissue plasminogen activator-related antigen in human plasma: crypticity phenomenon and relationship to plasma fibrinolysis

    International Nuclear Information System (INIS)

    Wun, T.C.; Capuano, A.

    1987-01-01

    A two-site immunoradiometric assay for tissue plasminogen activator (tPA) antigen has been developed using immunoaffinity purified antibody. Various treatments enhanced the detection of tPA antigen in the plasma samples. Maximum detection was obtained by acidification of plasma to pH 4.8 to 6.5 or addition of 0.5 mol/L of L-lysine or L-arginine. Acidification or addition of lysine to plasma is also required for maximum immunoadsorption of plasma tPA antigen on anti-tPA-Ig-sepharose. These results indicate that plasma tPA antigen is partially cryptic to antibody in untreated plasma. The plasma tPA antigen isolated by immunoadsorption of either untreated plasma or acidified plasma on anti-tPA-Ig-sepharose consists mainly of a 100-kd plasminogen activator species as determined by fibrin-agar zymography. The 100-kd activity is possibly a tPA:inhibitor complex. A standardized sample preparation method was conveniently adopted by mixing 3 vol of plasma and 1 vol of 2 mol/L of L-lysine for the assay. Reconstitution and recovery studies showed that the method is specific and permits full detection of both free tPA and tPA:inhibitor complex. The validity of the assay is further supported by the finding that the spontaneous plasma fibrinolysis previously demonstrated to be dependent on plasma tPA antigen is correlated with tPA antigen content. Using the standardized assay, we found that tPA antigen concentrations in 16 blood bank plasmas are equivalent to 3.7 to 20 ng of 60 kd tPA/mL. In all the plasma tested, more than half of the antigen is undetected unless the plasma is treated as described above

  7. Immunoradiometric quantitation of tissue plasminogen activator-related antigen in human plasma: crypticity phenomenon and relationship to plasma fibrinolysis

    Energy Technology Data Exchange (ETDEWEB)

    Wun, T.C.; Capuano, A.

    1987-05-01

    A two-site immunoradiometric assay for tissue plasminogen activator (tPA) antigen has been developed using immunoaffinity purified antibody. Various treatments enhanced the detection of tPA antigen in the plasma samples. Maximum detection was obtained by acidification of plasma to pH 4.8 to 6.5 or addition of 0.5 mol/L of L-lysine or L-arginine. Acidification or addition of lysine to plasma is also required for maximum immunoadsorption of plasma tPA antigen on anti-tPA-Ig-sepharose. These results indicate that plasma tPA antigen is partially cryptic to antibody in untreated plasma. The plasma tPA antigen isolated by immunoadsorption of either untreated plasma or acidified plasma on anti-tPA-Ig-sepharose consists mainly of a 100-kd plasminogen activator species as determined by fibrin-agar zymography. The 100-kd activity is possibly a tPA:inhibitor complex. A standardized sample preparation method was conveniently adopted by mixing 3 vol of plasma and 1 vol of 2 mol/L of L-lysine for the assay. Reconstitution and recovery studies showed that the method is specific and permits full detection of both free tPA and tPA:inhibitor complex. The validity of the assay is further supported by the finding that the spontaneous plasma fibrinolysis previously demonstrated to be dependent on plasma tPA antigen is correlated with tPA antigen content. Using the standardized assay, we found that tPA antigen concentrations in 16 blood bank plasmas are equivalent to 3.7 to 20 ng of 60 kd tPA/mL. In all the plasma tested, more than half of the antigen is undetected unless the plasma is treated as described above.

  8. Plasma nitriding monitoring reactor: A model reactor for studying plasma nitriding processes using an active screen

    Science.gov (United States)

    Hamann, S.; Börner, K.; Burlacov, I.; Spies, H.-J.; Strämke, M.; Strämke, S.; Röpcke, J.

    2015-12-01

    A laboratory scale plasma nitriding monitoring reactor (PLANIMOR) has been designed to study the basics of active screen plasma nitriding (ASPN) processes. PLANIMOR consists of a tube reactor vessel, made of borosilicate glass, enabling optical emission spectroscopy (OES) and infrared absorption spectroscopy. The linear setup of the electrode system of the reactor has the advantages to apply the diagnostic approaches on each part of the plasma process, separately. Furthermore, possible changes of the electrical field and of the heat generation, as they could appear in down-scaled cylindrical ASPN reactors, are avoided. PLANIMOR has been used for the nitriding of steel samples, achieving similar results as in an industrial scale ASPN reactor. A compact spectrometer using an external cavity quantum cascade laser combined with an optical multi-pass cell has been applied for the detection of molecular reaction products. This allowed the determination of the concentrations of four stable molecular species (CH4, C2H2, HCN, and NH3). With the help of OES, the rotational temperature of the screen plasma could be determined.

  9. Plasma nitriding monitoring reactor: A model reactor for studying plasma nitriding processes using an active screen

    International Nuclear Information System (INIS)

    Hamann, S.; Röpcke, J.; Börner, K.; Burlacov, I.; Spies, H.-J.; Strämke, M.; Strämke, S.

    2015-01-01

    A laboratory scale plasma nitriding monitoring reactor (PLANIMOR) has been designed to study the basics of active screen plasma nitriding (ASPN) processes. PLANIMOR consists of a tube reactor vessel, made of borosilicate glass, enabling optical emission spectroscopy (OES) and infrared absorption spectroscopy. The linear setup of the electrode system of the reactor has the advantages to apply the diagnostic approaches on each part of the plasma process, separately. Furthermore, possible changes of the electrical field and of the heat generation, as they could appear in down-scaled cylindrical ASPN reactors, are avoided. PLANIMOR has been used for the nitriding of steel samples, achieving similar results as in an industrial scale ASPN reactor. A compact spectrometer using an external cavity quantum cascade laser combined with an optical multi-pass cell has been applied for the detection of molecular reaction products. This allowed the determination of the concentrations of four stable molecular species (CH 4 , C 2 H 2 , HCN, and NH 3 ). With the help of OES, the rotational temperature of the screen plasma could be determined

  10. Plasma nitriding monitoring reactor: A model reactor for studying plasma nitriding processes using an active screen

    Energy Technology Data Exchange (ETDEWEB)

    Hamann, S., E-mail: hamann@inp-greifswald.de; Röpcke, J. [INP-Greifswald, Felix-Hausdorff-Str. 2, 17489 Greifswald (Germany); Börner, K.; Burlacov, I.; Spies, H.-J. [TU Bergakademie Freiberg, Institute of Materials Engineering, Gustav-Zeuner-Str. 5, 09599 Freiberg (Germany); Strämke, M.; Strämke, S. [ELTRO GmbH, Arnold-Sommerfeld-Ring 3, 52499 Baesweiler (Germany)

    2015-12-15

    A laboratory scale plasma nitriding monitoring reactor (PLANIMOR) has been designed to study the basics of active screen plasma nitriding (ASPN) processes. PLANIMOR consists of a tube reactor vessel, made of borosilicate glass, enabling optical emission spectroscopy (OES) and infrared absorption spectroscopy. The linear setup of the electrode system of the reactor has the advantages to apply the diagnostic approaches on each part of the plasma process, separately. Furthermore, possible changes of the electrical field and of the heat generation, as they could appear in down-scaled cylindrical ASPN reactors, are avoided. PLANIMOR has been used for the nitriding of steel samples, achieving similar results as in an industrial scale ASPN reactor. A compact spectrometer using an external cavity quantum cascade laser combined with an optical multi-pass cell has been applied for the detection of molecular reaction products. This allowed the determination of the concentrations of four stable molecular species (CH{sub 4}, C{sub 2}H{sub 2}, HCN, and NH{sub 3}). With the help of OES, the rotational temperature of the screen plasma could be determined.

  11. Purification and some properties of a novel maltohexaose-producing exo-amylase from Aerobacter aerogenes.

    Science.gov (United States)

    Kainuma, K; Wako, K; Kobayashi, S; Nogami, A; Suzuki, S

    1975-12-18

    Maltohexaose producing amylase (EC 3.2.1.-) is the fourth known exo-amylase, the three previously known being glucoamylase, beta-amylase and Pseudomonas stutzeri maltotetraose producing amylase. The enzyme after release from Aerobacter aerogenes cells by 0.1% sodium lauryl sulfate extraction was purified by ammonium sulfate precipitation, DEAE-Sephadex column chromatography and Sephadex G-100 gel filtration to 80-fold of the original sodium lauryl sulfate extract activity, It gave a single band on disc electrophoresis, and the molecular weight by gel filtration was 54 000. This amylase showed maximal activity at 50 degrees C and pH 6.80. The pH stability range was relatively wide, the enzyme retaining more than 90% of its initial activity in the range of 6.50-9.0. 80% of the activity was retained after 15 min at 50 degrees C. This enzyme produced maltohexaose from starch, amylose and amylopectin by exo-attack, but did not act on alpha- or beta-cyclodextrin, pullulan or maltohexaitol. Also the enzyme acted on beta-limit dextrins of amylopectin and glycogen to form branched oligosaccharides. The unusual reaction of this enzyme on beta-limit dextrin is discussed from the standpoint of the stereochemistry of 1,4-alpha- and 1,6-alpha-glucosidic bonds. This is the anomalous amylase for which it is recognized that 1,6-alpha-glucosidic linkages in the substrates can mimic the effect of 1,4-alpha-bonds, as previously observed in pseudo-priming reactions of E. coli phosphorylase.

  12. New insight into structure/function relationships in plant alpha-amylase family GH13 members

    DEFF Research Database (Denmark)

    Seo, Eun-Seong; Andersen, Joakim Mark; Nielsen, Morten Munch

    2010-01-01

    Two carbohydrate binding surface sites (SBSs) on barley α-amylase 1 (AMY1) of glycoside hydrolase family 13 (GH13) displayed synergy in interactions with starch granules, thus being pivotal for hydrolysis of supramolecular substrates. Mutational analysis showed that SBS1 is more critical for the ......Two carbohydrate binding surface sites (SBSs) on barley α-amylase 1 (AMY1) of glycoside hydrolase family 13 (GH13) displayed synergy in interactions with starch granules, thus being pivotal for hydrolysis of supramolecular substrates. Mutational analysis showed that SBS1 is more critical...... binding domains (SBDs) mediate binding to starch granules. SBDs are currently categorised into 9 carbohydrate binding module (CBM) families. A novel CBM20 subfamily encountered in regulatory enzymes possesses characteristically low affinity for β-CD. Although α-amylase is essential for starch mobilisation...... in germinating barley seeds, efficient degradation requires the concerted action of α-amylase, β-amylase, limit dextrinase (LD) and possibly α-glucosidase. Limit dextrinase (LD) is encoded by a single gene and represents the sole debranching activity during germination. Recent expression of functional LD...

  13. Occurrence of toxicity among protease, amylase, and color mutants of a nontoxic soy sauce koji mold

    International Nuclear Information System (INIS)

    Kalayanamitr, A.; Bhumiratana, A.; Flegel, T.W.; Glinsukon, T.; Shinmyo, A.

    1987-01-01

    A soy sauce koji mold, Aspergillus flavus var. columnaris Raper and Fennel (ATCC 44310), was treated with UV irradiation to obtain mutant strains possessing high protease activities, high amylase activities, and light-colored conidia. Selected mutant strains were tested for toxicity, and some were found acutely toxic to weanling rats, although all were negative for aflatoxin production

  14. Relation between salivary amylase and cortisol respnses to different stress tasks: Impact of sex

    NARCIS (Netherlands)

    van Stegeren, A.H.; Wolf, O.T.; Kindt, M.

    2008-01-01

    Neuro-endocrine markers such as salivary alpha amylase (sAA) and cortisol (CORT) play an important role in establishing human responses to stressful events. Whereas sAA levels reflect sympathetic system activity, salivary cortisol appears to be a valid measure for HPA axis activity. Although many

  15. Advanced qualification methodology for actively cooled plasma facing components

    Science.gov (United States)

    Durocher, A.; Escourbiac, F.; Grosman, A.; Boscary, J.; Merola, M.; Cismondi, F.; Courtois, X.; Farjon, J. L.; Missirlian, M.; Schlosser, J.; Tivey, R.

    2007-12-01

    The use of high heat flux plasma facing components (PFCs) in steady state fusion devices requires high reliability. These components have to withstand heat fluxes in the range 10-20 MW m-2 involving a number of severe engineering constraints. Feedback from the experience of various industrial manufacturings showed that the bonding of the refractory armour material onto the metallic heat sink causes generic difficulties strongly depending on material qualities and specific design. As the heat exhaust capability and lifetime of PFCs during plasma operation are directly linked to the manufacturing quality, a set of qualification activities such as active infrared thermography, lock-in and acoustic measurements were performed during the component development phases following a qualification route. This paper describes the major improvements stemming from better measurement accuracy and refined data processing and analyses recent developments aimed at investigating the capability to qualify the component in situ during its lifetime.

  16. Advanced qualification methodology for actively cooled plasma facing components

    International Nuclear Information System (INIS)

    Durocher, A.; Escourbiac, F.; Grosman, A.; Boscary, J.; Merola, M.; Cismondi, F.; Courtois, X.; Farjon, J.L.; Missirlian, M.; Schlosser, J.; Tivey, R.

    2007-01-01

    The use of high heat flux plasma facing components (PFCs) in steady state fusion devices requires high reliability. These components have to withstand heat fluxes in the range 10-20 MW m -2 involving a number of severe engineering constraints. Feedback from the experience of various industrial manufacturings showed that the bonding of the refractory armour material onto the metallic heat sink causes generic difficulties strongly depending on material qualities and specific design. As the heat exhaust capability and lifetime of PFCs during plasma operation are directly linked to the manufacturing quality, a set of qualification activities such as active infrared thermography, lock-in and acoustic measurements were performed during the component development phases following a qualification route. This paper describes the major improvements stemming from better measurement accuracy and refined data processing and analyses recent developments aimed at investigating the capability to qualify the component in situ during its lifetime

  17. Functional implications of plasma membrane condensation for T cell activation.

    Directory of Open Access Journals (Sweden)

    Carles Rentero

    2008-05-01

    Full Text Available The T lymphocyte plasma membrane condenses at the site of activation but the functional significance of this receptor-mediated membrane reorganization is not yet known. Here we demonstrate that membrane condensation at the T cell activation sites can be inhibited by incorporation of the oxysterol 7-ketocholesterol (7KC, which is known to prevent the formation of raft-like liquid-ordered domains in model membranes. We enriched T cells with 7KC, or cholesterol as control, to assess the importance of membrane condensation for T cell activation. Upon 7KC treatment, T cell antigen receptor (TCR triggered calcium fluxes and early tyrosine phosphorylation events appear unaltered. However, signaling complexes form less efficiently on the cell surface, fewer phosphorylated signaling proteins are retained in the plasma membrane and actin restructuring at activation sites is impaired in 7KC-enriched cells resulting in compromised downstream activation responses. Our data emphasizes lipids as an important medium for the organization at T cell activation sites and strongly indicates that membrane condensation is an important element of the T cell activation process.

  18. Growth temperature of different local isolates of Bacillus sp. in the solid state affects production of raw starch digesting amylases

    Directory of Open Access Journals (Sweden)

    Šokarda-Slavić Marinela

    2014-01-01

    Full Text Available Natural amylase producers, wild type strains of Bacillus sp., were isolated from different regions of Serbia. Strains with the highest amylase activity based on the starch-agar plate test were grown on solid-state fermentation (SSF on triticale. The influence of the substrate and different cultivation temperature (28 and 37°C on the production of amylase was examined. The tested strains produced α-amylases when grown on triticale grains both at 28 and at 37°C, but the activity of amylases and the number and intensity of the produced isoforms were different. Significant hydrolysis of raw cornstarch was obtained with the Bacillus sp. strains 2B, 5B, 18 and 24B. The produced α-amylases hydrolyzed raw cornstarch at a temperature below the temperature of gelatinization, but the ability for hydrolysis was not directly related to the total enzyme activity, suggesting that only certain isoforms are involved in the hydrolysis. [Projekat Ministarstva nauke Republike Srbije, br. 172048

  19. Polymeric amylase nanoparticles as a new semi-synthetic enzyme system for hydrolysis of starch.

    Science.gov (United States)

    Say, R; Şenay, R Hilal; Biçen, Özlem; Ersöz, Arzu; Şişman Yılmaz, Filiz; Akgöl, Sinan; Denizli, Adil

    2013-05-01

    α-Amylase (EC 3.2.1.1; α-D-1,4,glucan glucanohydrolase) catalyzes the hydrolysis of α-D-(1,4)-glucosidic linkages in starch, glycogen, and various malto-oligosaccharides, by releasing α-anomeric products. In this study, a novel method has been developed to prepare nanoprotein particles that carry α-amylase as a monomer by using a photosensitive microemulsion polymerization process. The nanostructured α-amylase with photosensitive features have been characterized by fluorescence spectroscopy, transmission electron microscopy (TEM) and Zeta Sizer. The fluorescence intensity of amylase nanoparticles was determined to be 658 a.u. at 610 nm and the average particle size of nanoamylase was found to be about 71.8 nm. Both free α-amylase and nanoparticles were used in the hydrolysis of starch under varying reaction conditions such as pH and temperature that affect enzyme activity and the results were compared to each other. Km values were 0.26 and 0.87 mM and Vmax values were 0.36 IU mg(-1) and 22.32 IU mg(-1) for nanoenzyme and free enzyme, respectively. Then, thermal stability, storage stability and reusability were investigated and according to the results, activity was preserved 60% at 60 °C; 20% at 70-80 °C temperature values and 80% after 105 days storage. Finally after 10 cycles, the activity was preserved 90% and this novel enzymatic polymeric amylase nanoparticle has showed considerable potential as reusable catalyst. Copyright © 2012 Elsevier B.V. All rights reserved.

  20. High yield of amylase from Aspergillus niger by the effect of gamma irradiation

    Energy Technology Data Exchange (ETDEWEB)

    El-Saadany, R.M.A.; Salem, F.A.; El-Manawaty, H.K.

    1984-02-01

    When irradiated rice was used as a media for Aspergillus niger a noticeable increase of amylase production was observed. Molecular degradation of starch molecules did occur, and an increase in starch acidity and solubility was noticed, whereas a marked decrease in viscosity as well as swelling capacity was observed. Gelatinization time and temperature of irradiated starch became shorter or lower resp. These results showed that internal changes in irradiated starch molecules and an alteration in its molecular configuration occured. They may affect the pathway of the growth of the fungi Aspergillus niger. When the amount of amylase was determined by measuring enzyme activity, it was observed that amylases in the irradiated media were higher than in the control media.

  1. A high yield of amylase from Aspergillus niger by the effect of gamma irradiation

    International Nuclear Information System (INIS)

    El-Saadany, R.M.A.; Salem, F.A.; El-Manawaty, H.K.

    1984-01-01

    When irradiated rice was used as a media for Aspergillus niger a noticeable increase of amylase production was observed. Molecular degradation of starch molecules did occure, and an increase in starch acidity and solubility was noticed, whereas a marked decrease in viscosity as well as swelling capacity was observed. Gelatinization time and temperature of irradiated starch became shorter or lower resp. These results showed that internal changes in irradiated starch molecules and an alteration in its molecular configuration occured. They may affect the pathway of the growth of the fungi Aspergillus niger. When the amount of amylase was determined by measuring enzyme activity, it was observed that amylases in the irradiated media were higher than in the control media. (orig.) [de

  2. Amylase-Binding Protein B of Streptococcus gordonii Is an Extracellular Dipeptidyl-Peptidase▿

    OpenAIRE

    Chaudhuri, Biswendu; Paju, Susanna; Haase, Elaine M.; Vickerman, M. Margaret; Tanzer, Jason M.; Scannapieco, Frank A.

    2008-01-01

    The oral commensal bacterium Streptococcus gordonii interacts with salivary amylase via two amylase-binding proteins, AbpA and AbpB. Based on sequence analysis, the 20-kDa AbpA protein is unique to S. gordonii, whereas the 82-kDa AbpB protein appears to share sequence homology with other bacterial dipeptidases. The aim of this study was to verify the peptidase activity of AbpB and further explore its potential functions. The abpB gene was cloned, and histidine-tagged AbpB (His-AbpB) was expre...

  3. Supplementation of Carbohydrate to Enhance the α-amylase Production by Bacillus licheniformis ATCC 6346 in Presence of Seed Cakes

    Directory of Open Access Journals (Sweden)

    Vengadaramana, A.

    2012-01-01

    Full Text Available Aims: The effect of carbohydrate and amino acids on the production of a-amylase by Bacillus licheniformis ATCC 6346 was investigated. Methodology and results: To find out the influence of carbohydrate the total carbohydrate content of the medium containing different concentration (2-18 g/L of defatted seed cake powder of sesamum and mustard containing medium was kept constant by the addition of soluble starch separately. The highest a-amylase activity obtained in the medium containing 18g/L mustard (59.11+b1.48 U/mL and sesamum seed cake powder (55.23+b1.55 U/mL. The results indicated that under these conditions the carbohydrate content had no effect on the production of a-amylase. Effect of amino acids (0.2g/L of glycine, methionine, proline, lysine, leucine, threonine, serine, arginine, alanine, glutamic acid, tryptophan, glutamine, asparagine, histidine, valine, phenylalanine, isoleucine and mixture of amino acids on the production of a-amylase in fermentation medium was investigated. Among the different amino acids supplemented, eight amino acids improved the a-amylase production but casaminoacids slightly inhibited the enzyme production. In presence of tryptophan highest enzyme activity was obtained than control. Conclusion, significance and impact of study: In these study amino acids especially tryptophan takes part in a particular role rather than carbohydrate in the production of a-amylase from B. licheniformis ATCC 6346.

  4. Post-heparin plasma lipoprotein lipase, but not hepatic lipase activity, is related to plasma adiponectin in type 2 diabetic patients and healthy subjects

    NARCIS (Netherlands)

    De Vries, R; Wolffenbuttel, BHR; Sluiter, WJ; Van Tol, A; Dullaart, RPF

    2005-01-01

    The aim of this study was to determine the relationships of plasma adiponectin with post-heparin plasma lipoprotein lipase (LPL) and hepatic lipase (HL) activities, and to evaluate whether plasma adiponectin contributes to diabetes-associated dyslipidaemia. Plasma adiponectin, post-heparin plasma

  5. How to Patch Active Plasma and Collisionless Sheath: Practical Guide

    International Nuclear Information System (INIS)

    Kaganovich, Igor D.

    2002-01-01

    Most plasmas have a very thin sheath compared with the plasma dimension. This necessitates separate calculations of the plasma and sheath. The Bohm criterion provides the boundary condition for calculation of plasma profiles. To calculate sheath properties, a value of electric field at the plasma-sheath interface has to be specified in addition to the Bohm criterion. The value of the boundary electric field and robust procedure to approximately patch plasma and collisionless sheath with a very good accuracy are reported

  6. Investigating the Hydrolysis of Starch Using "a"-Amylase Contained in Dishwashing Detergent and Human Saliva

    Science.gov (United States)

    Munegumi, Toratane; Inutsuka, Masato; Hayafuji, Yukitaka

    2016-01-01

    Although saliva has commonly been used to teach about digestion by organisms, the phenomenon of digestion is actually caused by enzymes as catalytic substances. This activity explores the hydrolysis of starch by "a"-amylase in cleaning materials as well as a comparison with the similar reaction using human saliva. The fact that the…

  7. Isolation of α-amylase from malted rice (Wita 7) extract using ...

    African Journals Online (AJOL)

    The crude α-amylase from the 8th day malt extract was subsequently purified using starch column procedure. The purified extract recorded an estimated activity of 75.549 U/ml. The project further examined the effects of varying the ratios of starch adsorbent height to column diameter (L/D) during the purification step. For the ...

  8. Determinants of salivary evening alpha-amylase in a large sample free of psychopathology

    NARCIS (Netherlands)

    Veen, Gerthe; Giltay, Erik J.; Vreeburg, Sophie A.; Licht, Carmilla M. M.; Cobbaert, Christa M.; Zitman, Frans G.; Penninx, Brenda W. J. H.

    Objective: Recently, salivary alpha-amylase (sAA) has been proposed as a suitable index for sympathetic activity and dysregulation of the autonomic nervous system (ANS). Although determinants of sAA have been described, they have not been studied within the same study with a large sample size

  9. Interactions between Barley a-Amylases, Substrates, Inhibitors and Regulatory Proteins

    DEFF Research Database (Denmark)

    Hachem, Maher Abou; Bozonnet, Sophie; Willemoës, Martin

    2006-01-01

    Barley a-amylase binds sugars at two sites on the enzyme surface in addition to the active site. Crystallography and site-directed mutagenesis highlight the importance of aromatic residues at these surface sites as demonstrated by Kd values determined for ß-cyclodextrin by surface plasmon resonan...

  10. Characterization of a digestive α-amylase in the larvae of Pieris brassicae L. (Lepidoptera: Pieridae

    Directory of Open Access Journals (Sweden)

    Arash eZibaee

    2016-03-01

    Full Text Available The current study deals with a digestive α-amylase in the larvae of Pieris brassicae L. through purification, enzymatic characterization, gene expression and in vivo effect of a specific inhibitor, Acarbose. Although α-amylase activity was the highest in the whole gut homogenate of larvae but compartmentalization of amylolytic activity showed an equal activity in posterior midgut (PM and anterior midgut (AM. A three step purification using ammonium sulfate, Sepharyl G-100 and DEAE-Cellulose Fast flow revealed an enzyme with a specific activity of 5.18 U/mg, recovery of 13.20, purification fold of 19.25 and molecular weight of 88 kDa. The purified α-amylase had the highest activity at optimal pH and temperature of 8 and 35 ºC. Also, the enzyme had Vmax values of 4.64 and 3.02 U/mg protein and Km values of 1.37 and 1.74% using starch and glycogen as substrates, respectively. Different concentrations of acarbose, ethylenediamine tetraacetic acid and ethylene glycol-bis (β-aminoethylether N, N, N′, N′-tetraacetic acid significantly decreased activity of the purified α-amylase. The 4th instar larvae of P. brassicae were fed on the treated leaves of Raphanus sativus L. with 0.22 mM of Acarbose to find in vivo effects on nutritional indices, α-amylase activity and gene expression. The significant differences were only found in conversion efficiency of digested food, relative growth rate and metabolic cost of control and fed larvae on Acarbose. Also, amylolytic activity significantly decreased in the treated larvae by both biochemical and native-PAGE experiments. Results of RT-PCR revealed a gene with 621 bp length responsible for α-amylase expression that had 75% identity with Papilio xuthus and P. polytes. Finally, qRT-PCR revealed higher expression of α-amylase in control larvae compared to acarbose-fed ones.

  11. Antidiabetic Indian Plants: A Good Source of Potent Amylase Inhibitors

    Directory of Open Access Journals (Sweden)

    Menakshi Bhat

    2011-01-01

    Full Text Available Diabetes is known as a multifactorial disease. The treatment of diabetes (Type II is complicated due to the inherent patho-physiological factors related to this disease. One of the complications of diabetes is post-prandial hyperglycemia (PPHG. Glucosidase inhibitors, particularly α-amylase inhibitors are a class of compounds that helps in managing PPHG. Six ethno-botanically known plants having antidiabetic property namely, Azadirachta indica Adr. Juss.; Murraya koenigii (L. Sprengel; Ocimum tenuflorum (L. (syn: Sanctum; Syzygium cumini (L. Skeels (syn: Eugenia jambolana; Linum usitatissimum (L. and Bougainvillea spectabilis were tested for their ability to inhibit glucosidase activity. The chloroform, methanol and aqueous extracts were prepared sequentially from either leaves or seeds of these plants. It was observed that the chloroform extract of O. tenuflorum; B. spectabilis; M. koenigii and S. cumini have significant α-amylase inhibitory property. Plants extracts were further tested against murine pancreatic, liver and small intestinal crude enzyme preparations for glucosidase inhibitory activity. The three extracts of O. tenuflorum and chloroform extract of M. koenigi showed good inhibition of murine pancreatic and intestinal glucosidases as compared with acarbose, a known glucosidase inhibitor.

  12. Photosynthesis Activates Plasma Membrane H+-ATPase via Sugar Accumulation.

    Science.gov (United States)

    Okumura, Masaki; Inoue, Shin-Ichiro; Kuwata, Keiko; Kinoshita, Toshinori

    2016-05-01

    Plant plasma membrane H(+)-ATPase acts as a primary transporter via proton pumping and regulates diverse physiological responses by controlling secondary solute transport, pH homeostasis, and membrane potential. Phosphorylation of the penultimate threonine and the subsequent binding of 14-3-3 proteins in the carboxyl terminus of the enzyme are required for H(+)-ATPase activation. We showed previously that photosynthesis induces phosphorylation of the penultimate threonine in the nonvascular bryophyte Marchantia polymorpha However, (1) whether this response is conserved in vascular plants and (2) the process by which photosynthesis regulates H(+)-ATPase phosphorylation at the plasma membrane remain unresolved issues. Here, we report that photosynthesis induced the phosphorylation and activation of H(+)-ATPase in Arabidopsis (Arabidopsis thaliana) leaves via sugar accumulation. Light reversibly phosphorylated leaf H(+)-ATPase, and this process was inhibited by pharmacological and genetic suppression of photosynthesis. Immunohistochemical and biochemical analyses indicated that light-induced phosphorylation of H(+)-ATPase occurred autonomously in mesophyll cells. We also show that the phosphorylation status of H(+)-ATPase and photosynthetic sugar accumulation in leaves were positively correlated and that sugar treatment promoted phosphorylation. Furthermore, light-induced phosphorylation of H(+)-ATPase was strongly suppressed in a double mutant defective in ADP-glucose pyrophosphorylase and triose phosphate/phosphate translocator (adg1-1 tpt-2); these mutations strongly inhibited endogenous sugar accumulation. Overall, we show that photosynthesis activated H(+)-ATPase via sugar production in the mesophyll cells of vascular plants. Our work provides new insight into signaling from chloroplasts to the plasma membrane ion transport mechanism. © 2016 American Society of Plant Biologists. All Rights Reserved.

  13. Improved α-amylase production by Aspergillus oryzae after a double deletion of genes involved in carbon catabolite repression.

    Science.gov (United States)

    Ichinose, Sakurako; Tanaka, Mizuki; Shintani, Takahiro; Gomi, Katsuya

    2014-01-01

    In filamentous fungi, the expression of secretory glycoside hydrolase encoding genes, such as those for amylases, cellulases, and xylanases, is generally repressed in the presence of glucose. CreA and CreB have been observed to be regulating factors for carbon catabolite repression. In this study, we generated single and double deletion creA and/or creB mutants in Aspergillus oryzae. The α-amylase activities of each strain were compared under various culture conditions. For the wild-type strain, mRNA levels of α-amylase were markedly decreased in the later stage of submerged culture under inducing conditions, whereas this reduced expression was not observed for single creA and double creA/creB deletion mutants. In addition, α-amylase activity of the wild-type strain was reduced in submerged culture containing high concentrations of inducing sugars, whereas all constructed mutants showed higher α-amylase activities. In particular, the α-amylase activity of the double deletion mutant in a medium containing 5% starch was >10-fold higher than that of the wild-type strain under the same culture conditions. In solid-state cultures using wheat bran as a substrate, the α-amylase activities of single creA and double deletion mutants were >2-fold higher than that of the wild-type strain. These results suggested that deleting both creA and creB resulted in dramatic improvements in the production of secretory glycoside hydrolases in filamentous fungi.

  14. Influence of media composition on the production of alkaline α-amylase from Bacillus subtilis CB-18.

    Science.gov (United States)

    Ogbonnaya, Nwokoro; Odiase, Anthonia

    2012-01-01

    Starch, a homopolysaccharide is an important and an abundant food reserve and energy source. Starches are processed to yield different products which find many industrial applications. Alpha-amylases hydrolyze starch by cleaving α-1,4-glucosidic bonds and have been used in food, textile and pharmaceutical industries [Sun et al. 2010]. Enzymatic conversion of starch with amylase presents an economically superior alternative to the conventional method of starch gelatinization. Alkaline α-amylase has an important position in the global enzyme market as a constituent of detergent. In this paper, we screened soil bacteria and an isolate, alkalophilic Bacillus subtilis CB-18 was found to produce an alkaline α-amylase in different media. MATERIAL AND METHODS. Screening of the isolates for amylolytic activity was carried out by growing bacteria isolated from the soil in starch agar plates and subsequently staining the plates with iodine solution to reveal zones of hydrolysis of starch. The selected isolate, Bacillus subtlis CB-18 was grown in different media at alkaline pH to evaluate the influence of media composition on alkaline α-amylase production. Enzyme assay was carried out by growing the culture in a broth medium and obtaining cell - free culture supernatant after centrifugation at 2515 × g for 15 minutes Amylase activity was determined by incubating 0.5 ml of crude enzyme solution in 0.1M Tris/HCl buffer (pH 8.5) with 0.5 ml of 1% soluble starch solution. The reaction was terminated by the addition of DNS reagent and reducing sugar produced from the amylolytic reaction was determined. Bacillus subtilis CB-18 used for this work was selected because it produced 7 mm zone diameter on starch agar plate. This organism was cultured in different alkaline broth media containing 2% soluble starch as inducer carbohydrate for α-amylase production. Among the carbon sources used for enzyme production, sorbitol was the best to stimulate enzyme production with α-amylase

  15. Molecular cloning of alpha-amylases from cotton boll weevil, Anthonomus grandis and structural relations to plant inhibitors: an approach to insect resistance.

    Science.gov (United States)

    Oliveira-Neto, Osmundo B; Batista, João A N; Rigden, Daniel J; Franco, Octávio L; Falcão, Rosana; Fragoso, Rodrigo R; Mello, Luciane V; dos Santos, Roseane C; Grossi-de-Sá, Maria F

    2003-01-01

    Anthonomus grandis, the cotton boll weevil, causes severe cotton crop losses in North and South America. Here we demonstrate the presence of starch in the cotton pollen grains and young ovules that are the main A. grandis food source. We further demonstrate the presence of alpha-amylase activity, an essential enzyme of carbohydrate metabolism for many crop pests, in A. grandis midgut. Two alpha-amylase cDNAs from A. grandis larvae were isolated using RT-PCR followed by 5' and 3' RACE techniques. These encode proteins with predicted molecular masses of 50.8 and 52.7kDa, respectively, which share 58% amino acid identity. Expression of both genes is induced upon feeding and concentrated in the midgut of adult insects. Several alpha-amylase inhibitors from plants were assayed against A. grandis alpha-amylases but, unexpectedly, only the BIII inhibitor from rye kernels proved highly effective, with inhibitors generally active against other insect amylases lacking effect. Structural modeling of Amylag1 and Amylag2 showed that different factors seem to be responsible for the lack of effect of 0.19 and alpha-AI1 inhibitors on A. grandis alpha-amylase activity. This work suggests that genetic engineering of cotton to express alpha-amylase inhibitors may offer a novel route to A. grandis resistance.

  16. Partial purification and characterization of {alpha}-amylases from one insecticide-resistant population of Sitophilus zeamais

    Energy Technology Data Exchange (ETDEWEB)

    Lopes, K.V.; Oliveira, M.G.A.; Paixao, G.P.; Visotto, L.E. [Universidade Federal de Vicosa (UFV), MG (Brazil). Dept. de Bioquimica e Biologia Molecular; Veloso, R.V.S.; Marinho, J.S.; Guedes, R.N.C. [Universidade Federal de Vicosa (UFV), MG (Brazil). Dept. de Biologia Animal; Oliveira, J.A. [Universidade Federal de Vicosa (UFV), MG (Brazil). Dept. de Quimica

    2008-07-01

    Full text: {alpha}-Amylases (EC 3.2.1.1) constitute a family of endo-amylases that catalyze the hydrolysis of a-D- (1,4)-glucan linkages in st ach components and various other related carbohydrates. They play a central role in carbohydrate metabolism of animals, plants and microorganisms. Many insects, especially those that feed on grain products during larval and/or adult life, depend on their amylases for survival. This is particularly true for the Sitophilus zeamais Motschulsky, a cosmopolitan pest of stored products. It is mainly controlled by insecticides. Amylases from adults of S.zeamais insecticide-resistant were purified by using a sequential procedure of glycogen-complex precipitation and ion exchange chromatography. Specific activity increased from 58,0454 AU/dL/mg protein in the crude homogenate to 2558,8720 AU/dL/mg protein in the final purified sample. Amylase unit (AU/dL) refers to the amount of amylase that hydrolysis 10 mg starch in 30 min at 37 deg C. The purified amylase ran as a single protein band on SDS-PAGE. From a plot of log molecular weight against relative mobility in 10% acrylamide gel, molecular weight was estimated to be 56 kDa. The enzyme had a K{sub m} of 0,2243 g/L for soluble starch and was most active at ph 5,0. The temperature of major activity was 40 deg C. The activity of enzyme was unaffected by presence or absence of Cl{sup -} and Ca{sup 2+}.

  17. Partial purification and characterization of α-amylases from one insecticide-resistant population of Sitophilus zeamais

    International Nuclear Information System (INIS)

    Lopes, K.V.; Oliveira, M.G.A.; Paixao, G.P.; Visotto, L.E.; Veloso, R.V.S.; Marinho, J.S.; Guedes, R.N.C.; Oliveira, J.A.

    2008-01-01

    Full text: α-Amylases (EC 3.2.1.1) constitute a family of endo-amylases that catalyze the hydrolysis of a-D- (1,4)-glucan linkages in st ach components and various other related carbohydrates. They play a central role in carbohydrate metabolism of animals, plants and microorganisms. Many insects, especially those that feed on grain products during larval and/or adult life, depend on their amylases for survival. This is particularly true for the Sitophilus zeamais Motschulsky, a cosmopolitan pest of stored products. It is mainly controlled by insecticides. Amylases from adults of S.zeamais insecticide-resistant were purified by using a sequential procedure of glycogen-complex precipitation and ion exchange chromatography. Specific activity increased from 58,0454 AU/dL/mg protein in the crude homogenate to 2558,8720 AU/dL/mg protein in the final purified sample. Amylase unit (AU/dL) refers to the amount of amylase that hydrolysis 10 mg starch in 30 min at 37 deg C. The purified amylase ran as a single protein band on SDS-PAGE. From a plot of log molecular weight against relative mobility in 10% acrylamide gel, molecular weight was estimated to be 56 kDa. The enzyme had a K m of 0,2243 g/L for soluble starch and was most active at ph 5,0. The temperature of major activity was 40 deg C. The activity of enzyme was unaffected by presence or absence of Cl - and Ca 2+

  18. Computer modeling of active experiments in space plasmas

    International Nuclear Information System (INIS)

    Bollens, R.J.

    1993-01-01

    The understanding of space plasmas is expanding rapidly. This is, in large part, due to the ambitious efforts of scientists from around the world who are performing large scale active experiments in the space plasma surrounding the earth. One such effort was designated the Active Magnetospheric Particle Tracer Explorers (AMPTE) and consisted of a series of plasma releases that were completed during 1984 and 1985. What makes the AMPTE experiments particularly interesting was the occurrence of a dramatic anomaly that was completely unpredicted. During the AMPTE experiment, three satellites traced the solar-wind flow into the earth's magnetosphere. One satellite, built by West Germany, released a series of barium and lithium canisters that were detonated and subsequently photo-ionized via solar radiation, thereby creating an artificial comet. Another satellite, built by Great Britain and in the vicinity during detonation, carried, as did the first satellite, a comprehensive set of magnetic field, particle and wave instruments. Upon detonation, what was observed by the satellites, as well as by aircraft and ground-based observers, was quite unexpected. The initial deflection of the ion clouds was not in the ambient solar wind's flow direction (rvec V) but rather in the direction transverse to the solar wind and the background magnetic field (rvec V x rvec B). This result was not predicted by any existing theories or simulation models; it is the main subject discussed in this dissertation. A large three dimensional computer simulation was produced to demonstrate that this transverse motion can be explained in terms of a rocket effect. Due to the extreme computer resources utilized in producing this work, the computer methods used to complete the calculation and the visualization techniques used to view the results are also discussed

  19. Progress of pancreatitis disease biomarker alpha amylase enzyme by new nano optical sensor.

    Science.gov (United States)

    Attia, M S; Al-Radadi, Najlaa S

    2016-12-15

    A new nano optical sensor binuclear Pd-(2-aminothiazole) (urea), Pd(atz,ur) complex was prepared and characterized for the assessment of the activity of alpha amylase enzyme in urine and serum samples for early diagnosis of Pancreatitis disease. The assessment of alpha amylase activity is carried out by the quenching of the luminescence intensity of the nano optical sensor binuclear Pd(atz,ur) complex at 457nm by the 2-chloro-4-nitrophenol (2-CNP) which produced from the reaction of the enzyme with 2-chloro-4-nitrophenyl-α-d-maltotrioside (CNPG3) substrate. The remarkable quenching of the luminescence intensity at 457nm of nano Pd(atz,ur) doped in sol-gel matrix by various concentrations of the 2-CNP was successfully used as an optical sensor for the assessment of α-amylase activity. The calibration plot was achieved over the concentration range 8.5×10(-6) to 1.9×10(-9)molL(-1) 2-CNP with a correlation coefficient of (0.999) and a detection limit of (7.4×10(-10)molL(-1)). The method was used satisfactorily for the assessment of the α-amylase activity over activity range (3-321U/L) in different urine and serum samples of pancreatitis patients. The assessment of the alpha amylase biomarker by the proposed method increases its sensitivity (96.88%) and specificity (94.41%) for early diagnosis of pancreatitis diseases. Copyright © 2016 Elsevier B.V. All rights reserved.

  20. Plasma B-esterase activities in European raptors.

    Science.gov (United States)

    Roy, Claudie; Grolleau, Gérard; Chamoulaud, Serge; Rivière, Jean-Louis

    2005-01-01

    B-esterases are serine hydrolases composed of cholinesterases, including acetylcholinesterase (AChE) and butyrylcholinesterase (BChE), and carboxylesterase (CbE). These esterases, found in blood plasma, are inhibited by organophosphorus (OP) and carbamate (CB) insecticides and can be used as nondestructive biomarkers of exposure to anticholinesterase insecticides. Furthermore, B-esterases are involved in detoxification of these insecticides. In order to establish the level of these enzymes and to have reference values for their normal activities, total plasma cholinesterase (ChE), AChE and BChE activities, and plasma CbE activity were determined in 729 European raptors representing 20 species, four families, and two orders. The diurnal families of the Falconiforme order were represented by Accipitridae and Falconidae and the nocturnal families of the Strigiforme order by Tytonidae and Strigidae. Intraspecies differences in cholinesterase activities according to sex and/or age were investigated in buzzards (Buteo buteo), sparrowhawks (Accipiter nisus), kestrels (Falco tinnunculus), barn owls (Tyto alba), and tawny owls (Strix aluco). Sex-related differences affecting ChE and AChE activities were observed in young kestrels (2-3-mo-old) and age-related differences in kestrels (ChE and AChE), sparrowhawks (AChE), and tawny owls (ChE, AChE, and BChE). The interspecies analysis yielded a negative correlation between ChE activity and body mass taking into account the relative contribution of AChE and BChE to ChE activity, with the exception of the honey buzzard (Pernis apivorus). The lowest ChE activities were found in the two largest species, Bonelli's eagle (Hieraaetus fasciatus) and Egyptian vulture (Neophron percnopterus) belonging to the Accipitridae family. The highest ChE activities were found in the relatively small species belonging to the Tytonidae and Strigidae families and in honey buzzard of the Accipitridae family. Species of the Accipitridae, Tytonidae, and

  1. Atividade da amilase no intestino delgado de coelhos alimentados com dietas contendo diferentes níveis de amido antes e após desmame = Amylase activity in the small intestine of rabbits fed with diets containing starch levels before and after weaning

    Directory of Open Access Journals (Sweden)

    Haroldo Garcia de Faria

    2006-07-01

    Full Text Available Para avaliar a atividade amilásica em coelhos após a desmama, foram utilizados 200 animais da Raça Nova Zelândia Branco, submetidos a diferentes padrões de alimentação desde a fase de lactação. Quarenta deles foram abatidos aos 35 dias de idade e 80, aos 50 e aos 70 dias de idade. Para modular o padrão de consumo de leite e ração antes da desmama, as ninhadas equalizadas em 8 láparos do nascimento até ao 16º dia de idade, a partir desta idade, até a desmama, foram submetidos à alta e baixa disponibilidade de leite, mantendo ninhadas com 4 ou 8 láparos recebendo dietas com nível normal (15% e alto (24% de amido. A partir da desmama, os animais de cada um dos 4 padrões de alimentação até a desmama passaram a receber as mesmasdietas com níveis normal e alto de amido, em um esquema fatorial 2x2x2 (ninhadas submetidas à baixa e alta disponibilidade de leite versus dois níveis de amido na dieta até a desmama versus doisníveis de amido na dieta após a desmama. Os resultados mostraram que a manutenção de ninhadas com 4 ou 8 animais entre o 16º e 35º dia de lactação não modificou (P>0,05 a atividade da amilase em qualquer das idades avaliadas após o desmame. O uso de dietas com alto teor de amido durante a lactação aumentou (POne experiment was carried out to study the amylase activity in the small intestine (jejunum and ileum of rabbits from litters maintained on different feeding patterns. Two hundred White New Zealand rabbits were analyzed, out of which 40 35- days-old rabbits, 80 50-days-old rabbits and 80 70-days-old rabbits were slaughtered. To modulate the pattern of feed and milk intake before weaning, the litters were modulated for eight young rabbits until 16 days old. From 16 days until weaning, they were submitted to low or high milkavailability, litters maintained with four or eight young rabbits receiving diets containing normal or high starch level. The animals were distributed in a factorial design 2

  2. High quality actively cooled plasma facing components for fusion

    International Nuclear Information System (INIS)

    Nygren, R.

    1993-01-01

    This paper interweaves some suggestions for developing actively-cooled PFCs (plasma facing components) for future fusion devices with supporting examples taken from the design, fabrication and operation of Tore Supra's Phase III Outboard Pump Limiter (OPL). This actively-cooled midplane limiter, designed for heat and particle removal during long pulse operation, has been operated in essentially thermally steady state conditions. From experience with testing to identify braze flaws in the OPL, recommendations are made to analyze the impact of joining flaws on thermal-hydraulic performance of PFCs and to validate a method of inspection for such flaws early in the design development. Capability for extensive in-service monitoring of future PFCs is also recommended and the extensive calorimetry and IR thermography used to confirm and update safe operating limits for power handling of the OPL are reviewed

  3. Enhanced amylase production by fusarium solani in solid state fermentation

    International Nuclear Information System (INIS)

    Bakri, Y.; Jawhar, M.; Arabi, M.I.E.

    2014-01-01

    The present study illustrates the investigation carried out on the production of amylase by Fusarium species under solid state fermentation. All the tested Fusarium species were capable of producing amylase. A selected F. solani isolate SY7, showed the highest amylase production in solid state fermentation. Different substrates were screened for enzyme production. Among the several agronomic wastes, wheat bran supported the highest yield of amylase (141.18 U/g of dry substrate) after 3 days of incubation. Optimisation of the physical parameters revealed the optimum pH, temperature and moisture level for amylase production by the isolate as 8.0, 25 C and 70%, respectively. The above results indicate that the production of amylase by F. solani isolate SY7 could be improved by a further optimisation of the medium and culture conditions. (author)

  4. Synchronization to light and mealtime of daily rhythms of locomotor activity, plasma glucose and digestive enzymes in the Nile tilapia (Oreochromis niloticus).

    Science.gov (United States)

    Guerra-Santos, Bartira; López-Olmeda, José Fernando; de Mattos, Bruno Olivetti; Baião, Alice Borba; Pereira, Denise Soledade Peixoto; Sánchez-Vázquez, Francisco Javier; Cerqueira, Robson Bahia; Albinati, Ricardo Castelo Branco; Fortes-Silva, Rodrigo

    2017-02-01

    The light-dark cycle and feeding can be the most important factors acting as synchronizers of biological rhythms. In this research we aimed to evaluate synchronization to feeding schedule of daily rhythms of locomotor activity and digestive enzymes of tilapia. For that purpose, 120 tilapias (65.0±0.6g) were distributed in 12 tanks (10 fish per tank) and divided into two groups. One group was fed once a day at 11:00h (zeitgeber time, ZT6) (ML group) and the other group was fed at 23:00h (ZT18) (MD group). The fish were anesthetized to collect samples of blood, stomach and midgut at 4-hour intervals over a period of 24h. Fish fed at ML showed a diurnal locomotor activity (74% of the total daily activity occurring during the light phase) and synchronization to the feeding schedule, as this group showed anticipation to the feeding time. Fish fed at MD showed a disruption in the pattern of locomotor activity and became less diurnal (59%). Alkaline protease activity in the midgut showed daily rhythm with the achrophase at the beginning of the dark phase in both ML and MD groups. Acid protease and amylase did not show significant daily rhythms. Plasma glucose showed a daily rhythm with the achrophase shifted by 12h in the ML and MD groups. These results revealed that the feeding time and light cycle synchronize differently the daily rhythms of behavior, digestive physiology and plasma metabolites in the Nile tilapia, which indicate the plasticity of the circadian system and its synchronizers. Copyright © 2016 Elsevier Inc. All rights reserved.

  5. Attenuation of Porphyromonas gingivalis oral infection by α-amylase and pentamidine.

    Science.gov (United States)

    Li, Ying; Miao, Yu-Song; Fu, Yun; Li, Xi-Ting; Yu, Shao-Jie

    2015-08-01

    The Porphyromonas gingivalis bacterium is one of the most influential pathogens in oral infections. In the current study, the antimicrobial activity of α-amylase and pentamidine against Porphyromonas gingivalis was evaluated. Their in vitro inhibitory activity was investigated with the agar overlay technique, and the minimal inhibitory and bactericidal concentrations were determined. Using the bactericidal concentration, the antimicrobial actions of the inhibitors were investigated. In the present study, multiple techniques were utilized, including scanning electron microscopy (SEM), general structural analysis and differential gene expression analysis. The results obtained from SEM and bactericidal analysis indicated a notable observation; the pentamidine and α-amylase treatment destroyed the structure of the bacterial cell membranes, which led to cell death. These results were used to further explore these inhibitors and the mechanisms by which they act. Downregulated expression levels were observed for a number of genes coding for hemagglutinins and gingipains, and various genes involved in hemin uptake, chromosome replication and energy production. However, the expression levels of genes associated with iron storage and oxidative stress were upregulated by α-amylase and pentamidine. A greater effect was noted in response to pentamidine treatment. The results of the present study demonstrate promising therapeutic potential for α-amylases and pentamidine. These molecules have the potential to be used to develop novel drugs and broaden the availability of pharmacological tools for the attenuation of oral infections caused by Porphyromonas gingivalis.

  6. Production and immobilization of alpha amylase using biotechnology techniques for use in biological and medical applications

    International Nuclear Information System (INIS)

    Mobasher, E.E.F.

    2009-01-01

    The immobilized enzymes on polymeric supports are prepared for purpose of repeated use and the possibilities of continuous reaction system. One of the most important properties is the stability of proteins when they are used in some medical and industrial applications. The immobilization of the enzymes improves this property as well as many other properties.In this study, alpha amylase was purified and immobilized onto two different polymers. α- amylase was used in this study for its biological and industrial applications. It is used in paper textile, pharmaceutical applications, food, and detergent industries. α- amylase was found in plants, animals, and microorganisms. Purification of α-amylase from microorganisms is the main source of α-amylase because it was excreted from many bacteria and fungi. In this study, α-amylase was purified from Aspergillus niger. Fractional precipitation of the α- amylase produced by A. niger with 80% ammonium sulphate saturation. The crude enzyme was applied on column chromatography packed with Sephadex G 100 for purification. The active eluents containing partially purified enzyme were collected for further investigation. The specific activity of α-amylase was (34.9 U/mg) which was corresponding to 2.09 fold purification for the tested organism. The purified α-amylase was immobilized by entrapment method into two types of polymers. One of them was natural consist of chitosan and alginate. The other polymer was synthetic consist of N- isopropyl acrylamide and alginate. The temperature optimum and thermal inactivation showed a severe loss in the activity of the free enzymes, while the temperature profile of the immobilized enzymes was much broader at higher temperatures demonstrating the effectiveness of the polymer protecting the enzymes. Also, the immobilized enzymes (natural polymer and synthetic polymer) showed higher thermal stability. Optimum ph and stability showed that immobilization of enzymes resulted in more

  7. Fluorogenic MMP activity assay for plasma including MMPs complexed to α2-macroglobulin

    NARCIS (Netherlands)

    Beekman, B.; Drijfhout, J.W.; Ronday, H.K.; TeKoppele, J.M.

    1999-01-01

    Elevated MMP activities are implicated in tissue degradation in, e.g., arthritis and cancer. The present study was designed to measure MMP enzyme activity in plasma. Free active MMP is unlikely to be present in plasma: upon entering the circulation, active MMP is expected to be captured by the

  8. Science and technology of plasma activated direct wafer bonding

    Science.gov (United States)

    Roberds, Brian Edward

    This dissertation studied the kinetics of silicon direct wafer bonding with emphasis on low temperature bonding mechanisms. The project goals were to understand the topological requirements for initial bonding, develop a tensile test to measure the bond strength as a function of time and temperature and, using the kinetic information obtained, develop lower temperature methods of bonding. A reproducible surface metrology metric for bonding was best described by power spectral density derived from atomic force microscopy measurements. From the tensile strength kinetics study it was found that low annealing temperatures could be used to obtain strong bonds, but at the expense of longer annealing times. Three models were developed to describe the kinetics. A diffusion controlled model and a reaction rate controlled model were developed for the higher temperature regimes (T > 600sp°C), and an electric field assisted oxidation model was proposed for the low temperature range. An in situ oxygen plasma treatment was used to further enhance the field-controlled mechanism which resulted in dramatic increases in the low temperature bonding kinetics. Multiple internal transmission Fourier transform infrared spectroscopy (MIT-FTIR) was used to monitor species evolution at the bonded interface and a capacitance-voltage (CV) study was undertaken to investigate charge distribution and surface states resulting from plasma activation. A short, less than a minute, plasma exposure prior to contacting the wafers was found to obtain very strong bonds for hydrophobic silicon wafers at very low temperatures (100sp°C). This novel bonding method may enable new technologies involving heterogeneous material systems or bonding partially fabricated devices to become realities.

  9. REB-instability with magneto-active inhomogeneous warm plasma

    International Nuclear Information System (INIS)

    El-Shorbagy, K.H.

    2000-07-01

    The beam-plasma heating due to a relativistic electron beam (REB) under the effect of an external static magnetic field is investigated. It is considered that a longitudinal 1-D oscillations exist in the plasma, which is inhomogeneous and bounded in the direction of the beam propagation. It is found that the variation in the plasma density has a profound effect on the spatial beam-plasma instability. Besides, the external static magnetic field and warmness of plasma electron leads to more power absorption from the electron beam, and consequently an auxiliary plasma heating. (author)

  10. REB-Instability with Magneto-Active Inhomogeneous Warm Plasma

    International Nuclear Information System (INIS)

    El-Shorbagy, Kh.H.

    2000-01-01

    The beam-plasma heating due to a relativistic electron beam (REB) under the effect of an external static magnetic field is investigated. It is considered that a longitudinal 1-D oscillations exist in the plasma, which is inhomogeneous and bounded in the direction of the beam propagation. It is found that the variation in the plasma density has a profound effect on the spatial beam-plasma instability. Besides, the external static magnetic field and warmness of plasma electron leads to more power absorption from the electron beam, and consequently an auxiliary plasma heating

  11. Bottleneck in secretion of α-amylase in Bacillus subtilis.

    Science.gov (United States)

    Yan, Shaomin; Wu, Guang

    2017-07-19

    Amylase plays an important role in biotechnology industries, and Gram-positive bacterium Bacillus subtilis is a major host to produce heterogeneous α-amylases. However, the secretion stress limits the high yield of α-amylase in B. subtilis although huge efforts have been made to address this secretion bottleneck. In this question-oriented review, every effort is made to answer the following questions, which look simple but are long-standing, through reviewing of literature: (1) Does α-amylase need a specific and dedicated chaperone? (2) What signal sequence does CsaA recognize? (3) Does CsaA require ATP for its operation? (4) Does an unfolded α-amylase is less soluble than a folded one? (5) Does α-amylase aggregate before transporting through Sec secretion system? (6) Is α-amylase sufficient stable to prevent itself from misfolding? (7) Does α-amylase need more disulfide bonds to be stabilized? (8) Which secretion system does PrsA pass through? (9) Is PrsA ATP-dependent? (10) Is PrsA reused after folding of α-amylase? (11) What is the fate of PrsA? (12) Is trigger factor (TF) ATP-dependent? The literature review suggests that not only the most of those questions are still open to answers but also it is necessary to calculate ATP budget in order to better understand how B. subtilis uses its energy for production and secretion.

  12. Plasma composition in a sigmoidal anemone active region

    International Nuclear Information System (INIS)

    Baker, D.; Van Driel-Gesztelyi, L.; Green, L. M.; Carlyle, J.; Brooks, D. H.; Démoulin, P.; Steed, K.

    2013-01-01

    Using spectra obtained by the EUV Imaging Spectrometer (EIS) instrument onboard Hinode, we present a detailed spatially resolved abundance map of an active region (AR)-coronal hole (CH) complex that covers an area of 359'' × 485''. The abundance map provides first ionization potential (FIP) bias levels in various coronal structures within the large EIS field of view. Overall, FIP bias in the small, relatively young AR is 2-3. This modest FIP bias is a consequence of the age of the AR, its weak heating, and its partial reconnection with the surrounding CH. Plasma with a coronal composition is concentrated at AR loop footpoints, close to where fractionation is believed to take place in the chromosphere. In the AR, we found a moderate positive correlation of FIP bias with nonthermal velocity and magnetic flux density, both of which are also strongest at the AR loop footpoints. Pathways of slightly enhanced FIP bias are traced along some of the loops connecting opposite polarities within the AR. We interpret the traces of enhanced FIP bias along these loops to be the beginning of fractionated plasma mixing in the loops. Low FIP bias in a sigmoidal channel above the AR's main polarity inversion line, where ongoing flux cancellation is taking place, provides new evidence of a bald patch magnetic topology of a sigmoid/flux rope configuration.

  13. Actively cooled plasma facing components qualification, commissioning and health monitoring

    International Nuclear Information System (INIS)

    Escourbiac, F.; Durocher, A.; Grosman, A.; Courtois, X.; Farjon, J.-L.; Schlosser, J.; Merola, M.; Tivey, R.

    2006-01-01

    In modern steady state magnetic fusion devices, actively cooled plasma facing components (PFC) have to handle heat fluxes in the range of 10-20 MW/m 2 . This generates a number of engineering constraints: the armour materials must be refractory and compatible with plasma wall interaction requirements (low sputtering and/or low atomic number); the heat sink must offer high thermal conductivity, high mechanical resistance and sufficient ductility; the component cooling system -which is generally based on the circulation of pressurized water in the PFC's heat sink - must offer high thermal heat transfer efficiency. Furthermore, the assembling of the refractory armour material onto the metallic heat sink causes generic difficulties strongly depending on thermo-mechanical properties of materials and design requirements. Life time of the PFC during plasma operation are linked to their manufacturing quality, in particular they are reduced by the possible presence of flaw assembling. The fabrication of PFC in an industrial frame including their qualification and their commissioning - which consists in checking the manufacturing quality during and at the end of manufacture - is a real challenge. From experience gained at Tore Supra on carbon fibre composite flat tiles technology components, it was assessed that a set of qualifications activities must be operated during R(and)D and manufacturing phases. Dedicated Non Destructive Technique (NDT) based on advanced active infrared thermography was developed for this purpose, afterwards, correlations between NDT, high heat flux testing and thermomechanical modelling were performed to analyse damage detection and propagation, and define an acceptance criteria valuable for industrial application. Health monitoring using lock-in technique was also recently operated in-situ of the Tore Supra tokamak for detection of possible defect propagation during operations, presence of acoustic precursor for critical heat flux detection induced

  14. An exceptionally cold-adapted alpha-amylase from a metagenomic library of a cold and alkaline environment

    DEFF Research Database (Denmark)

    Vester, Jan Kjølhede; Glaring, Mikkel Andreas; Stougaard, Peter

    2015-01-01

    A cold-active α-amylase, AmyI3C6, identified by a functional metagenomics approach was expressed in Escherichia coli and purified to homogeneity. Sequence analysis showed that the AmyI3C6 amylase was similar to α-amylases from the class Clostridia and revealed classical characteristics of cold......-adapted enzymes, as did comparison of the kinetic parameters Km and kcat to a mesophilic α-amylase. AmyI3C6 was shown to be heat-labile. Temperature optimum was at 10-15 °C, and more than 70 % of the relative activity was retained at 1 °C. The pH optimum of AmyI3C6 was at pH 8-9, and the enzyme displayed activity...... in two commercial detergents tested, suggesting that the AmyI3C6 α-amylase may be useful as a detergent enzyme in environmentally friendly, low-temperature laundry processes....

  15. Microencapsulation of Purified Amylase Enzyme from Pitaya (Hylocereus polyrhizus Peel in Arabic Gum-Chitosan using Freeze Drying

    Directory of Open Access Journals (Sweden)

    Mehrnoush Amid

    2014-03-01

    Full Text Available Amylase is one of the most important enzymes in the world due to its wide application in various industries and biotechnological processes. In this study, amylase enzyme from Hylocereus polyrhizus was encapsulated for the first time in an Arabic gum-chitosan matrix using freeze drying. The encapsulated amylase retained complete biocatalytic activity and exhibited a shift in the optimum temperature and considerable increase in the pH and temperature stabilities compared to the free enzyme. Encapsulation of the enzyme protected the activity in the presence of ionic and non-ionic surfactants and oxidizing agents (H2O2 and enhanced the shelf life. The storage stability of amylase is found to markedly increase after immobilization and the freeze dried amylase exhibited maximum encapsulation efficiency value (96.2% after the encapsulation process. Therefore, the present study demonstrated that the encapsulation of the enzyme in a coating agent using freeze drying is an efficient method to keep the enzyme active and stable until required in industry.

  16. Synthesis and in vitro study of benzofuran hydrazone derivatives as novel alpha-amylase inhibitor.

    Science.gov (United States)

    Taha, Muhammad; Shah, Syed Adnan Ali; Imran, Syahrul; Afifi, Muhammad; Chigurupati, Sridevi; Selvaraj, Manikandan; Rahim, Fazal; Ullah, Hayat; Zaman, Khalid; Vijayabalan, Shantini

    2017-12-01

    The α-amylase acts as attractive target to treat type-2 diabetes mellitus. Therefore in discovering a small molecule as α-amylase inhibitor, we have synthesized benzofuran carbohydrazide analogs (1-25), characterized through different spectroscopic techniques such as 1 HNMR and EI-MS. All screened analog shows good α-amylase inhibitory potentials with IC 50 value ranging between 1.078±0.19 and 2.926±0.05µM when compared with acarbose having IC 50 =0.62±0.22µM. Only nine analogs among the series such as analogs 3, 5, 7, 8, 10, 12, 21, 23 and 24 exhibit good inhibitory potential with IC 50 values 1.644±0.128, 1.078±0.19, 1.245±0.25, 1.843±0.19, 1.350±0.24, 1.629±0.015, 1.353±0.232, 1.359±0.119 and 1.488±0.07µM when compare with standard drug acarbose. All other analogs showed good to moderate α-amylase inhibitory potentials. The SAR study was conducted on the basis of substituent difference at the phenyl ring. The binding interaction between analogs and active site of enzyme was confirmed by docking studies. Copyright © 2017 Elsevier Inc. All rights reserved.

  17. Michaelis kinetic analysis of extracellular cellulase and amylase excreted by Lactobacillus plantarum during cassava fermentation

    Science.gov (United States)

    Frediansyah, Andri; Kurniadi, Muhamad

    2017-01-01

    Our previous study reveal that single culture of Lactobacillus plantarum has ability to ferment cassava tuber in relation to produce modified cassava flour (mocaf). It was used to accelerate a fermentation process. L. plantarum grow well and produce some extracellular enzymes i.e. cellulase to change the structure and breakdown the cell wall of cassava tuber. Then, the starchy materials will be hydrolyzed by i.e. amylase into simple sugar and convert to organic acid. All of these process will give new characteristic of cassava i.e. lower fiber content, good flavor, taste, aroma and texture and the amount of cyanide acid is lower. Therefore this present study was to analyze Michaelis kinetics of extracellular carboxymethyl cellulase and amylase production by L. plantarum during cassava fermentation. The maximum carboxymethyl cellulase and amylase activity of 8.60 U/ml and 14.07 U/ml, respectively, were obtained from filtrate which has been incubated at 37°C for 18 h under stationary conditions. The Vmax and Km of CMCase were 0.8506 × 10-3 U/ml and 0.9594 × 10-3 g/mL, respectively. For amylase were 9.291 × 10-3 U/ml and 0.9163 × 10-3 g/ml, respectively.

  18. Functional characterization and crystal structure of thermostable amylase from Thermotoga petrophila, reveals high thermostability and an unusual form of dimerization

    DEFF Research Database (Denmark)

    Hameed, Uzma; Price, Ian; Ikram-Ul-Haq

    2017-01-01

    and able to hydrolyze starch into dextrin between 90 and 100°C, with optimum activity at 98°C and pH8.5. The activity increased in the presence of Rb(1+), K(1+) and Ca(2+) ions, whereas other ions inhibited activity. The crystal structure of Tp-AmyS at 1.7Å resolution showed common features of the GH-13...... of salivary amylase from a previous crystal structure, and thus could be a functional feature of some amylases....

  19. Activities report of the National Space Research Institute Plasma Laboratory for the period 1988/1989

    International Nuclear Information System (INIS)

    Ludwig, Gerson Otto.

    1990-11-01

    This report describes the activities performed in the period 1988/1989 by the National Space Research Institute (INPE/SCT) Plasma Laboratory (LAP). The report presents the main results in the following research lines: plasma physics, plasma technology, and controlled thermonuclear fusion. (author). 49 figs., 3 tabs

  20. Plasma renin activity in patients with ischaemic heart disease

    International Nuclear Information System (INIS)

    Urbanek, J.; Hofman, O.; Reisenauer, R.; Slaby, A.

    1977-01-01

    Plasma renin activity (PRA) stimulated by upright posture was measured in 300 men aged 45-64 years using a radioimmunoassay of angiotensin-I. The examined subjects were normotensive or patients with benign essential hypertension and were divided into 6 groups according to the absence of manifest atherosclerosis, the presence of definite angina pectoris or a history of myocardial infarction. Each group contained 50 unselected subjects, with a comparable mean age. Significant differences in mean PRA were found between corresponding groups of hypertensives and normotensives, the values in hypertensives being lower. The percentage of low renin values was higher in hypertensives with ischaemic heart disease than in other groups. It is suggested that this finding might be explained by functional disturbances in the kidneys in hypertensives with ischaemic heart disease. (orig.) [de

  1. Plasma renin activity in patients with ischaemic heart disease

    Energy Technology Data Exchange (ETDEWEB)

    Urbanek, J; Hofman, O; Reisenauer, R; Slaby, A [Karlova Universita, Prague (Czechoslovakia). Inst. of Biophysics and Nuclear Medicine; Karlova Universita, Prague (Czechoslovakia). IV. Dept. of Internal Medicine; Vyzkumny Ustav Endokrinologicky, Prague [Czechoslovakia

    1977-04-01

    Plasma renin activity (PRA) stimulated by upright posture was measured in 300 men aged 45 to 64 years using a radioimmunoassay of angiotensin-I. The examined subjects were normotensive or patients with benign essential hypertension and were divided into 6 groups according to the absence of manifest atherosclerosis, the presence of definite angina pectoris or a history of myocardial infarction. Each group contained 50 unselected subjects, with a comparable mean age. Significant differences in mean PRA were found between corresponding groups of hypertensives and normotensives, the values in hypertensives being lower. The percentage of low renin values was higher in hypertensives with ischaemic heart disease than in other groups. It is suggested that this finding might be explained by functional disturbances in the kidneys in hypertensives with ischaemic heart disease.

  2. Effect of gas pressure on active screen plasma nitriding response

    International Nuclear Information System (INIS)

    Nishimoto, Akio; Nagatsuka, Kimiaki; Narita, Ryota; Nii, Hiroaki; Akamatsu, Katsuya

    2010-01-01

    An austenitic stainless steel AISI 304 was active screen plasma nitrided using a 304 steel screen to investigate the effect of the gas pressure on the ASPN response. The sample was treated for 18 ks at 723 K in 25% N2 + 75% H2 gases. The gas pressure was changed to 100, 600 and 1200 Pa. The distance between screen and sample was also changed to 10, 30 and 50 mm. The nitrided samples were characterized by appearance observation, surface roughness, optical microscopy, X-ray diffraction, and microhardness testing. After nitriding, polygonal particles with a normal distribution were observed at the center and edges of all the ASPN-treated sample surfaces. Particles on the sample surfaces were finer with an increase in the gas pressure. The nitrided layer with a greater and homogeneous thickness was obtained at a low gas pressure of 100 Pa. (author)

  3. Beam-plasma coupling physics in support of active experiments

    Science.gov (United States)

    Yakymenko, K.; Delzanno, G. L.; Roytershteyn, V.

    2017-12-01

    The recent development of compact relativistic accelerators might open up a new era of active experiments in space, driven by important scientific and national security applications. Examples include using electron beams to trace magnetic field lines and establish causality between physical processes occurring in the magnetosphere and those in the ionosphere. Another example is the use of electron beams to trigger waves in the near-Earth environment. Waves could induce pitch-angle scattering and precipitation of energetic electrons, acting as an effective radiation belt remediation scheme. In this work, we revisit the coupling between an electron beam and a magnetized plasma in the framework of linear cold-plasma theory. We show that coupling can occur through two different regimes. In the first, a non-relativistic beam radiates through whistler waves. This is well known, and was in fact the focus of many rockets and space-shuttle campaigns aimed at demonstrating whistler emissions in the eighties. In the second regime, the beam radiates through extraordinary (R-X) modes. Nonlinear simulations with a highly-accurate Vlasov code support the theoretical results qualitatively and demonstrate that the radiated power through R-X modes can be much larger than in the whistler regime. Test-particle simulations in the wave electromagnetic field will also be presented to assess the efficiency of these waves in inducing pitch-angle scattering via wave-particle interactions. Finally, the implications of these results for a rocket active experiment in the ionosphere and for a radiation belt remediation scheme will be discussed.

  4. The effects of residual platelets in plasma on plasminogen activator inhibitor-1 and plasminogen activator inhibitor-1-related assays.

    Directory of Open Access Journals (Sweden)

    Marlien Pieters

    Full Text Available Due to controversial evidence in the literature pertaining to the activity of plasminogen activator inhibitor-1 in platelets, we examined the effects of residual platelets present in plasma (a potential pre-analytical variable on various plasminogen activator inhibitor-1 and plasminogen activator inhibitor-1-related assays. Blood samples were collected from 151 individuals and centrifuged at 352 and 1500 g to obtain plasma with varying numbers of platelet. In a follow-up study, blood samples were collected from an additional 23 individuals, from whom platelet-poor (2000 g, platelet-containing (352 g and platelet-rich plasma (200 g were prepared and analysed as fresh-frozen and after five defrost-refreeze cycles (to determine the contribution of in vitro platelet degradation. Plasminogen activator inhibitor-1 activity, plasminogen activator inhibitor-1 antigen, tissue plasminogen activator/plasminogen activator inhibitor-1 complex, plasma clot lysis time, β-thromboglobulin and plasma platelet count were analysed. Platelet α-granule release (plasma β-thromboglobulin showed a significant association with plasminogen activator inhibitor-1 antigen levels but weak associations with plasminogen activator inhibitor-1 activity and a functional marker of fibrinolysis, clot lysis time. Upon dividing the study population into quartiles based on β-thromboglobulin levels, plasminogen activator inhibitor-1 antigen increased significantly across the quartiles while plasminogen activator inhibitor-1 activity and clot lysis time tended to increase in the 4th quartile only. In the follow-up study, plasma plasminogen activator inhibitor-1 antigen was also significantly influenced by platelet count in a concentration-dependent manner. Plasma plasminogen activator inhibitor-1 antigen levels increased further after complete platelet degradation. Residual platelets in plasma significantly influence plasma plasminogen activator inhibitor-1 antigen levels mainly

  5. Isolation, purification and characterization of β-amylase from Dioscorea hispida Dennst

    Science.gov (United States)

    Oktiarni, Dwita; Lusiana, Simamora, Febri Yanti; Gaol, Jusni M. Lumban

    2015-09-01

    β-amylase (E.C 3.2.1.2) is an enzyme commonly found in plants and bacteria. The enzyme is an exo-acting carbohydrolase which hydrolyzes α-1.4-glucosidic linkages of starch, removing maltose units from the non-reducing end of the polysaccharide chain, producing β-maltose and β-limit dextrin as the final product. β-amylase is widely distributed in the higher plants such as sweet potato. Besides the use in starch hydrolysis, starch-converting enzymes are also used in a number of other industrial applications, such as laundry and porcelain detergents or as anti-stalling agents in baking. This enzyme was extracted from Dioscorea hispida Dennst in 0.05 M acetate buffer pH 4.8 and followed by ammonium sulfate fractionation at cold temperature (10°C). Ammonium sulfate fractionation was shared into fraction of 0-60%, 60-70%, 70-80% and 80-100%. The fraction containing high of specific activity (determined by Somogyi-Nelson and Lowry methods) was futher purified by dialysis. Fraction with high enzyme activity of β-amylase were fraction 60-70% and 70-80%, with specific activity of Dioscorea hispida Dennst were 1.32 and 1.55 mg sugar.mg protein-1.minute-1, whereas specific activity of crude extract enzyme was 0.21 mg sugar.mg protein-1.minute-1. After purified with dialysis, fraction with high enzyme activity of β-amylase were fraction of 60-70% and 70-80%, with specific activity of Dioscorea hispida Dennst was 2.72 and 4.24 mg sugar.mg protein-1.minute-1. The purified Dioscorea hispida Dennst β-amylase from dialysis showed increasing in spesific activity the crude enzyme as much as 24 folds. The characterization of enzyme showed that Dioscorea hispida Dennst derived enzyme had optimum pH of 5.5 and temperature of 70°C. The kinetic parameters of purified Dioscorea hispida Dennst β-amylase showed that the KMapp, Vmaxapp value and Hill constant were 0.0211 mg/ml, 9.63 mg sugar.minute-1 and 1.34, respectively.

  6. Plasma cholinesterase activity of rats, western grey kangaroos, alpacas, sheep, cattle, and horses.

    Science.gov (United States)

    Mayberry, Chris; Mawson, Peter; Maloney, Shane K

    2015-01-01

    Plasma cholinesterase activity levels of various species may be of interest to toxicologists or pathologists working with chemicals that interfere with the activity of plasma cholinesterase. We used a pH titration method to measure the plasma cholinesterase activity of six mammalian species. Plasma cholinesterase activity varied up to 50-fold between species: sheep (88 ± 45 nM acetylcholine degraded per ml of test plasma per minute), cattle (94 ± 35), western grey kangaroos (126 ± 92), alpaca (364 ± 70), rats (390 ± 118) and horses (4539 ± 721). We present a simple, effective technique for the assay of plasma cholinesterase activity levels from a range of species. Although labour-intensive, it requires only basic laboratory equipment. Copyright © 2015 Elsevier Inc. All rights reserved.

  7. Cow dung is an ideal fermentation medium for amylase production in solid-state fermentation by Bacillus cereus

    Directory of Open Access Journals (Sweden)

    Ponnuswamy Vijayaraghavan

    2015-12-01

    Full Text Available Amylase production by Bacillus cereus IND4 was investigated by solid state fermentation (SSF using cow dung substrate. The SSF conditions were optimized by using one-variable-at-a-time approach and two level full factorial design. Two level full factorial design demonstrated that moisture, pH, fructose, yeast extract and ammonium sulphate have significantly influenced enzyme production (p < 0.05. A central composite design was employed to investigate the optimum concentration of these variables affecting amylase production. Maximal amylase production of 464 units/ml of enzyme was observed in the presence of 100% moisture, 0.1% fructose and 0.01% ammonium sulphate. The enzyme production increased three fold compared to the original medium. The optimum pH and temperature for the activity of amylase were found to be 8.0 and 50 °C, respectively. This enzyme was highly stable at wide pH range (7.0–9.0 and showed 32% enzyme activity after initial denaturation at 50 °C for 1 h. This is the first detailed report on the production of amylase by microorganisms using cow dung as the low cost medium.

  8. Enhanced Production and Characterization of a Solvent Stable Amylase from Solvent Tolerant Bacillus tequilensis RG-01: Thermostable and Surfactant Resistant

    Directory of Open Access Journals (Sweden)

    Soni Tiwari

    2014-01-01

    Full Text Available Ten bacterial strains isolated from the soil samples in the presence of cyclohexane were screened for amylase production. Among them, culture RG-01 was adjudged as the best amylase producer and was identified as Bacillus tequilensis from MTCC, Chandigarh. The isolate showed maximum amylase production (8100 U/mL in the presence of starch, peptone, and Ca2+ ions at 55°C pH 7.0 within 24 h of incubation. The enzyme was stable in the presence of n-dodecane, isooctane, n-decane, xylene, toluene, n-hexane, n-butanol, and cyclohexane, respectively. The presence of benzene, methanol, and ethanol marginally reduced the amylase stability, respectively. The enzyme was showed it 100% activity at 55°C and pH 7.0 with 119% and 127% stability at 55°C and pH 7.0, respectively. The enzyme was also stable in the presence of SDS, Tween-40, Tween-60, and Tween-80 (1% and was found stimulatory effect, respectively. Only Triton-X-100 showed a moderate inhibitory effect (5% on amylase activity. This isolate (Bacillus tequilensis RG-01 may be useful in several industrial applications owing to its thermotolerant and organic solvents and surfactants resistance characteristics.

  9. Molecular cloning and characterization of an α-amylase cDNA highly expressed in major feeding stages of the coffee berry borer, Hypothenemus hampei.

    Science.gov (United States)

    Bezerra, C A; Macedo, L L P; Amorim, T M L; Santos, V O; Fragoso, R R; Lucena, W A; Meneguim, A M; Valencia-Jimenez, A; Engler, G; Silva, M C M; Albuquerque, E V S; Grossi-de-Sa, M F

    2014-12-10

    α-Amylases are common enzymes responsible for hydrolyzing starch. Insect-pests, whose larvae develop in seeds, rely obligatorily on α-amylase activity to digest starch, as their major food source. Considering the relevance of insect α-amylases and the natural α-amylase inhibitors present in seeds to protect from insect damage, we report here the molecular cloning and nucleotide sequence of the full-length AmyHha cDNA of the coffee berry borer, Hypothenemus hampei, a major insect-pest of coffee crops. The AmyHha sequence has 1879 bp, containing a 1458 bp open reading frame, which encodes a predicted protein with 485 amino acid residues, with a predicted molecular mass of 51.2 kDa. The deduced protein showed 55-79% identity to other insect α-amylases, including Anthonomus grandis, Ips typographus and Sitophilus oryzae α-amylases. In depth analysis revealed that the highly conserved three amino acid residues (Asp184, Glu220, and Asp285), which compose the catalytic site are also presented in AmyHha amylase. The AmyHha gene seems to be a single copy in the haploid genome and AmyHha transcription levels were found higher in L2 larvae and adult insects, both corresponding to major feeding phases. Modeling of the AmyHha predicted protein uncovered striking structural similarities to the Tenebrio molitor α-amylase also displaying the same amino acid residues involved in enzyme catalysis (Asp184, Glu220 and Asp285). Since AmyHha gene was mostly transcribed in the intestinal tract of H. hampei larvae, the cognate α-amylase could be considered a high valuable target to coffee bean insect control by biotechnological strategies. Copyright © 2014. Published by Elsevier B.V.

  10. Purification and Characterization of α-Amylase from Waste Bread ...

    African Journals Online (AJOL)

    M.Irshad

    2012-04-24

    Apr 24, 2012 ... The objective of this study was to purify and characterize the α-amylase for industrial perspective. The production of α-amylase through solid-state fermentation by Ganoderma tsuage was investigated by using waste bread as substrates. Production parameters were optimized as 2 mL of inoculum size,.

  11. Application of microbial α-amylase in industry - A review

    Directory of Open Access Journals (Sweden)

    Paula Monteiro de Souza

    2010-12-01

    Full Text Available Amylases are one of the main enzymes used in industry. Such enzymes hydrolyze the starch molecules into polymers composed of glucose units. Amylases have potential application in a wide number of industrial processes such as food, fermentation and pharmaceutical industries. α-Amylases can be obtained from plants, animals and microorganisms. However, enzymes from fungal and bacterial sources have dominated applications in industrial sectors. The production of α-amylase is essential for conversion of starches into oligosaccharides. Starch is an important constituent of the human diet and is a major storage product of many economically important crops such as wheat, rice, maize, tapioca, and potato. Starch-converting enzymes are used in the production of maltodextrin, modified starches, or glucose and fructose syrups. A large number of microbial α-amylases has applications in different industrial sectors such as food, textile, paper and detergent industries. The production of α-amylases has generally been carried out using submerged fermentation, but solid state fermentation systems appear as a promising technology. The properties of each α-amylase such as thermostability, pH profile, pH stability, and Ca-independency are important in the development of fermentation process. This review focuses on the production of bacterial and fungal α-amylases, their distribution, structural-functional aspects, physical and chemical parameters, and the use of these enzymes in industrial applications.

  12. Method for using a yeast alpha-amylase promoter

    Science.gov (United States)

    Gao, Johnway; Skeen, Rodney S.; Hooker, Brian S.; Anderson, Daniel B.

    2003-04-22

    The present invention provides the promoter clone discovery of an alpha-amylase gene of a starch utilizing yeast strain Schwanniomyces castellii. The isolated alpha-amylase promoter is an inducible promoter, which can regulate strong gene expression in starch culture medium.

  13. Expression and characterization of α-Amylases from penicillium ...

    African Journals Online (AJOL)

    In an attempt to enhance the industrial production of α-amylases in the tropics, sterile fresh bread was inoculated with spore suspensions of Penicillium citrinum at 25 oC. Extracellular α-amylases were produced and subjected to partial purification by ammonium sulphate precipitation and dialysis. Further purification by gel ...

  14. Purification and characterization of α-amylase from Ganoderma ...

    African Journals Online (AJOL)

    The objective of this study was to purify and characterize the α-amylase for industrial perspective. The production of α-amylase through solid-state fermentation by Ganoderma tsuage was investigated by using waste bread as substrates. Production parameters were optimized as 2 mL of inoculum size, moisture 50%, ...

  15. Application of microbial α-amylase in industry - A review.

    Science.gov (United States)

    de Souza, Paula Monteiro; de Oliveira Magalhães, Pérola

    2010-10-01

    Amylases are one of the main enzymes used in industry. Such enzymes hydrolyze the starch molecules into polymers composed of glucose units. Amylases have potential application in a wide number of industrial processes such as food, fermentation and pharmaceutical industries. α-Amylases can be obtained from plants, animals and microorganisms. However, enzymes from fungal and bacterial sources have dominated applications in industrial sectors. The production of α-amylase is essential for conversion of starches into oligosaccharides. Starch is an important constituent of the human diet and is a major storage product of many economically important crops such as wheat, rice, maize, tapioca, and potato. Starch-converting enzymes are used in the production of maltodextrin, modified starches, or glucose and fructose syrups. A large number of microbial α-amylases has applications in different industrial sectors such as food, textile, paper and detergent industries. The production of α-amylases has generally been carried out using submerged fermentation, but solid state fermentation systems appear as a promising technology. The properties of each α-amylase such as thermostability, pH profile, pH stability, and Ca-independency are important in the development of fermentation process. This review focuses on the production of bacterial and fungal α-amylases, their distribution, structural-functional aspects, physical and chemical parameters, and the use of these enzymes in industrial applications.

  16. Two Strategies for Microbial Production of an Industrial Enzyme-Alpha-Amylase

    Science.gov (United States)

    Bernhardsdotter, Eva C. M. J.; Garriott, Owen; Pusey, Marc L.; Ng, Joseph D.

    2003-01-01

    Extremophiles are microorganisms that thrive in, from an anthropocentric view, extreme environments including hot springs, soda lakes and arctic water. This ability of survival at extreme conditions has rendered extremophiles to be of interest in astrobiology, evolutionary biology as well as in industrial applications. Of particular interest to the biotechnology industry are the biological catalysts of the extremophiles, the extremozymes, whose unique stabilities at extreme conditions make them potential sources of novel enzymes in industrial applications. There are two major approaches to microbial enzyme production. This entails enzyme isolation directly from the natural host or creating a recombinant expression system whereby the targeted enzyme can be overexpressed in a mesophilic host. We are employing both methods in the effort to produce alpha-amylases from a hyperthermophilic archaeon (Thermococcus) isolated from a hydrothermal vent in the Atlantic Ocean, as well as from alkaliphilic bacteria (Bacillus) isolated from a soda lake in Tanzania. Alpha-amylases catalyze the hydrolysis of internal alpha-1,4-glycosidic linkages in starch to produce smaller sugars. Thermostable alpha-amylases are used in the liquefaction of starch for production of fructose and glucose syrups, whereas alpha-amylases stable at high pH have potential as detergent additives. The alpha-amylase encoding gene from Thermococcus was PCR amplified using carefully designed primers and analyzed using bioinformatics tools such as BLAST and Multiple Sequence Alignment for cloning and expression in E.coli. Four strains of Bacillus were grown in alkaline starch-enriched medium of which the culture supernatant was used as enzyme source. Amylolytic activity was detected using the starch-iodine method.

  17. Engineering high α-amylase levels in wheat grain lowers Falling Number but improves baking properties.

    Science.gov (United States)

    Ral, Jean-Philippe; Whan, Alex; Larroque, Oscar; Leyne, Emmett; Pritchard, Jeni; Dielen, Anne-Sophie; Howitt, Crispin A; Morell, Matthew K; Newberry, Marcus

    2016-01-01

    Late maturity α-amylase (LMA) and preharvest sprouting (PHS) are genetic defects in wheat. They are both characterized by the expression of specific isoforms of α-amylase in particular genotypes in the grain prior to harvest. The enhanced expression of α-amylase in both LMA and PHS results in a reduction in Falling Number (FN), a test of gel viscosity, and subsequent downgrading of the grain, along with a reduced price for growers. The FN test is unable to distinguish between LMA and PHS; thus, both defects are treated similarly when grain is traded. However, in PHS-affected grains, proteases and other degradative process are activated, and this has been shown to have a negative impact on end product quality. No studies have been conducted to determine whether LMA is detrimental to end product quality. This work demonstrated that wheat in which an isoform α-amylase (TaAmy3) was overexpressed in the endosperm of developing grain to levels of up to 100-fold higher than the wild-type resulted in low FN similar to those seen in LMA- or PHS-affected grains. This increase had no detrimental effect on starch structure, flour composition and enhanced baking quality, in small-scale 10-g baking tests. In these small-scale tests, overexpression of TaAmy3 led to increased loaf volume and Maillard-related browning to levels higher than those in control flours when baking improver was added. These findings raise questions as to the validity of the assumption that (i) LMA is detrimental to end product quality and (ii) a low FN is always indicative of a reduction in quality. This work suggests the need for a better understanding of the impact of elevated expression of specific α-amylase on end product quality. © 2015 The Authors. Plant Biotechnology Journal published by Society for Experimental Biology and The Association of Applied Biologists and John Wiley & Sons Ltd.

  18. Unripe Fruit's Extract of Quince (Cydonia oblonga Miller as a Potent Alpha-amylase Inhibitor

    Directory of Open Access Journals (Sweden)

    Mostafa Koutb

    2012-01-01

    Full Text Available The use of alpha-amylase inhibitors has recently gained in popularity with the success and growth of carbohydrate restricted diets. In this study, two different stages from the unripe fruits of quince (Cydonia oblonga Miller have been tested for their potentiality in alpha-amylase inhibition as a key enzyme in carbohydrates assimilation. Our results revealed that addition of different concentrations from extracts (0, 2, 4, 6, 8mg of dry mass of each stage of unripe fruits resulted in drastically decrease in the enzymatic activity of alpha-amylase by the percent of (0%, 42.6%, 21%, 26.3%, and 16.9% for the stage 1. Extracts from the stage 2 were more effective in enzymatic inhibition (0%, 26.9%, 3.8%, 0.2%, and 0.4%. The GC/MS analysis revealed that quince extract contains (sorbitol, quinic acid, p-vinylphenol and cyclopropane carboxylic acid. To explore which components are involved in the inhibition process, two pure components of the quince extract (sorbitol and quinic acid were used in inhibition assay. Neither sorbitol nor quinic acid shows any significant inhibition; therefore, these two components could be excluded from the inhibition process. Our current study suggested that p-vinylphenol and cyclopropane carboxylic acid might act as a-amylase inhibitors in vitro separately or synergistically. The possible explanation for the presence of cyclopropane carboxylic acid (CPCA in this critical phase of the unripe fruit will be discussed. This study suggests that the unripe fruits of quince can be used as a natural starch blocker containing alpha-amylase inhibitors which would be of interest for people requiring carbohydrate restricted diets.

  19. The design of actively cooled plasma-facing components

    International Nuclear Information System (INIS)

    Scheerer, M.; Smid, I.; Bolt, H.; Gervash, A.; Linke, J.

    2001-01-01

    In future fusion devices, like in the stellarator Wendelstein 7-X, the target plates of the divertor will be exposed to heat loads up to power densities of 10 MW/m 2 for 1000 s. For this purpose actively cooled target elements with an internal coolant flow return, made of 2-D CFC armor tiles brazed onto a two tube cooling structure were developed and manufactured at the Forschungszentrum Juelich. Individual bent- and coolant flow reversal elements were used to achieve a high flexibility in the shape of the target elements. A special brazing technology, using a thin layer of plasma-arc deposited titanium was used for the bonding of the cooling structure to the plasma facing armor (PFA). FEM-simulations of the thermal and mechanical behavior show that a detachment of about 25% of the bonded area between the copper tubes and the PFA can be tolerated, without exceeding the critical heat flux at 15 MW/m 2 or a surface temperature of 1400 C at 10 MW/m 2 by using twisted tape inserts with a twist ratio of 2 at a cooling water velocity of 10 m/s. Thermal cycling tests in an electron beam facility up to a power density level 10.5 MW/m 2 show a very good behavior of parts of the target elements, which confirms the performance under fusion relevant conditions. Even defected parts in the bonding interface of the target elements, known from ultrasonic inspections before, show no change in the thermal performance under cycling, which confirms also the structural integrity of partly defected regions. (orig.)

  20. Plasma Hazards and Acceptance for International Space Station Extravehicular Activities

    Science.gov (United States)

    Patton, Thomas

    2010-09-01

    Extravehicular activity(EVA) is accepted by NASA and other space faring agencies as a necessary risk in order to build and maintain a safe and efficient laboratory in space. EVAs are used for standard construction and as contingency operations to repair critical equipment for vehicle sustainability and safety of the entire crew in the habitable volume. There are many hazards that are assessed for even the most mundane EVA for astronauts, and the vast majority of these are adequately controlled per the rules of the International Space Station Program. The need for EVA repair and construction has driven acceptance of a possible catastrophic hazard to the EVA crewmember which cannot currently be controlled adequately. That hazard is electrical shock from the very environment in which they work. This paper describes the environment, causes and contributors to the shock of EVA crewmembers attributed to the ionospheric plasma environment in low Earth orbit. It will detail the hazard history, and acceptance process for the risk associated with these hazards that give assurance to a safe EVA. In addition to the hazard acceptance process this paper will explore other factors that go into the decision to accept a risk including criticality of task, hardware design and capability, and the probability of hazard occurrence. Also included will be the required interaction between organizations at NASA(EVA Office, Environments, Engineering, Mission Operations, Safety) in order to build and eventually gain adequate acceptance rationale for a hazard of this kind. During the course of the discussion, all current methods of mitigating the hazard will be identified. This paper will capture the history of the plasma hazard analysis and processes used by the International Space Station Program to formally assess and qualify the risk. The paper will discuss steps that have been taken to identify and perform required analysis of the floating potential shock hazard from the ISS environment

  1. A Study on Plasma Renin Activity in Essential Hypertension

    International Nuclear Information System (INIS)

    Choe, Kang Won; Lee, Jung Sang; Cho, Bo Yeon; Koh, Chang Soon; Lee, Mun Ho

    1975-01-01

    Radioimmunoassay for the measurement of plasma renin activity (PRA) was performed in 43 normal Koreans and 45 patients with essential hypertension. Plasma samples were drawn in supine position in the morning and after upright posture for 4 hours. Urinary sodium excretion rates were measured in the concurrent 24 hour urine samples, an index of their sodium balance. The results were as follows: 1) There was an inverse correlation between 24 hr sodium excretion and PRA. The normal values of PRA in supine position ranged from 1.0 to 7.0 ng/ml/hr. when 24 hour sodium excretion were between 50 to 150 mEq. PRA in elderly tended to be low. 2) When stimulated by 4 hour upright posture, PRA increased by 2.6 times from the baseline value. 3) Of the 45 patients with essential hypertension, PRA was low in 10 cases (22.2%), normal in 28 cases (62.2%), and high in 7 cases (15.6%). 4) In the normal and high renin groups, who tended to be younger in ages, mean diastolic blood pressure and BUN were higher than in low renin group. Though hypertensive retinopathy and left ventricular hypertrophy in ECG were more prevalent in the former, no significant differences were noted as in the case of serum cholesterol. 5) There were 8 cases of cardiovascular complications (7 with cerebral vascular accident, 1 with myocardial infarction); 3 in low renin group (30%), 2 in normal renin (7.1%) and 3 in higher renin group (42.9%). This figure indicated higher rate of cardiovascular complications in higher renin groups, and lower rate in normal renin group. But the incidence of the complication was not significantly low in low renin group.

  2. α-Amylase: an enzyme specificity found in various families of glycoside hydrolases

    DEFF Research Database (Denmark)

    Janeček, Štefan; Svensson, Birte; MacGregor, E. Ann

    2014-01-01

    of all carbohydrate-active enzymes, it is one of the most frequently occurring glycoside hydrolases (GH). α-Amylase is the main representative of family GH13, but it is probably also present in the families GH57 and GH119, and possibly even in GH126. Family GH13, known generally as the main α...... investigation because of an obvious, but unexpected, homology with inverting β-glucan-active hydrolases....

  3. Structural and regulatory differences in amylase isoenzymes in germinating Brazilian barley cultivars

    Directory of Open Access Journals (Sweden)

    Georg-Kraemer J.E.

    2003-01-01

    Full Text Available The amylase electrophoretic patterns of 10 Brazilian brewing-barley varieties with different malting grades and diastatic power were analyzed during the 7-day germination which occurs during the malting process. Intra and inter-variety genetic variability was observed at both the structural and regulatory level. In the first few days after germination all varieties showed a few active loci, all of them with low activity. In subsequent days, new loci became active and those already detected since early germination showed increased activity. All varieties showed a continuous increase in amylase synthesis until the 3rd and/or 4th day after germination. Some varieties maintained high amylase activity until the last day of germination, while others showed a decrease in activity on the 5th or 6th day. No specific band increased or decreased its intensity independently of the others. A total of 14 loci were detected, out of which only one locus was polymorphic, indicating very low structural genetic variability, with only 2.8% polymorphic loci, an average of 1.04 alleles per loci, and an average expected heterozygosity of only 0.7%. The mean intra-variety Jaccard similarity coefficient complement (1 - S J was 0.009. The mean intra-variety difference based on regulatory differences was higher (1 - S J = 0.17 than that obtained based on structural differences, suggesting differential gene activation. Inter-variety differentiation also showed low structural variability, with 1 - S J = 0.026 and a Nei genetic distance (D value of 0.0076, and a remarkable increase in divergence caused by differential gene activation (1 - S J = 0.34. These results indicate that regulatory polymorphism is the principal agent responsible for amylase variability in the barley varieties analyzed.

  4. Vacuum Plasma Spraying W-coated Reduced Activation Structural Steels for Fusion Plasma Facing Components

    Energy Technology Data Exchange (ETDEWEB)

    Noh, Sanghoon; Kim, Tae Kyu [Korea Atomic Energy Research Institute, Daejeon (Korea, Republic of)

    2014-05-15

    Tungsten (W) and its alloys are considered as candidate materials for plasma facing materials of the first wall and diverter components in fusion reactor systems because of high sputtering resistance and low tritium retention in a fusion environment. Therefore, it is considered that the joining between W and reduced activation structural steels, and its evaluation, are critical issues for the development of fusion reactors. However, the joining between these materials is a very challenging process because of significant differences in their physical properties, particularly the mismatch of coefficients of thermal expansion (CTE). For instance, the CTE of pure W is known to be about 4.3Χ10{sup -6}K{sup -1}; however, that of martensitic steels reaches over three times, about 12-14Χ10{sup -6}K{sup -1} at room temperature even up to 373K. Nevertheless, several joining techniques have been developed for joining between W and structural steels, such as a vapor deposition method, brazing and diffusion bonding. Meanwhile, vacuum plasma spraying (VPS) is supposed to be one of the prospective methods to fabricate a sufficient W layer on the steel substrates because of the coating of a large area with a relatively high fabricating rate. In this study, the VPS method of W powders on reduced activation steels was employed, and its microstructure and hardness distribution were investigated. ODS ferritic steels and F82H steel were coated by VPS-W, and the microstructure and hardness distribution were investigated. A microstructure analysis revealed that pure W was successfully coated on steel substrates by the VPS process without an intermediate layer, in spite of a mismatch of the CTE between dissimilar materials. After neutron irradiation, irradiation hardening significantly occurred in the VPSW. However, the hardening of VPS-W was lesser than that of bulk W irradiated HFIR at 773K. Substrate materials, ODS ferritic steels, and F82H steel, did not show irradiation hardening

  5. Efeitos de níveis de ácido L-glutâmico e de vitamina K da dieta sobre a atividade de alfa-amilase em frangos de corte Effects of dietary levels of L-glutamic acid and vitamin K in the activity of alpha-amylase of chicks

    Directory of Open Access Journals (Sweden)

    Flavia Escapini Fanchiotti

    2005-04-01

    Full Text Available Foram investigados os efeitos nutricionais de dois níveis de ácido L-glutâmico (L-Glu combinados com quatro níveis de vitamina K (Vit K sobre a atividade de alfa-amilase no quimo e pâncreas de aves de corte. Frangos de corte machos de um dia foram criados em baterias aquecidas e alimentados, à vontade, com dietas contendo todos L-aminoácidos essenciais, minerais e vitaminas (exceto Vit K até os 14 dias de idade. O experimento foi realizado em esquema fatorial, em delineamento inteiramente casualizado 2x4, com quatro repetições de oito aves cada. A dieta básica foi suplementada com 6,25 e 12,5% de L-Glu combinados com 0,02; 0,2; 20,0 e 200,0 mg de Vit K/kg de ração. Efeitos significativos de L-Glu e Vit K foram observados no quimo. A atividade específica máxima (1.005,78±245,25 UI/mg de proteína foi observada nas aves alimentadas com 6,25% de L-Glu e 20,0 mg de Vit K. Houve redução da atividade com a suplementação de 12,5% de L-Glu. No pâncreas, não houve interação entre L-Glu e Vit K, todavia, foi observado efeito de L-Glu sobre as atividades relativas, expressas em UI/g de tecido e UI/100 g de peso corporal. Os resultados sugerem que a associação entre L-Glu e Vit K interfere na atividade enzimática de alfa-amilase em aves de corte submetidas a dietas purificadas. Os resultados indicam que o nível de 12,5% de L-Glu associado aos diferentes níveis de vitamina K reduziu a atividade enzimática.The effects of two levels of L-glutamic acid (L-Glu combined with four levels of vitamin K (Vit K were studied with the objective of evaluating the activities of alpha-amylase in the chyme and pancreas of chicks. Day-old male broilers were reared in eletrically heated batteries, fed with purified amino acids diets, minerals and vitamins (except Vit K at the requirement levels, for 14 days. Feed and water were provided ad libitum. The experimental design was a factorial 2x4 with four replicates with eight chicks each The

  6. Microbial amylases in the production of alcohol

    Energy Technology Data Exchange (ETDEWEB)

    Pieper, H J

    1970-01-01

    This book is based on experiments carried out in the experimental distillery of the University of Hohenheim on the use of microbial enzyme preparations for processing wheat and maize, with particular reference to comparison of green and cured malts. The subject is divided into the following chapters: introduction (pp. -14); raw materials (pp. 5-6); enzymic dextrinizing and saccharification agents (pp. 6-10); technology of alcohol production with microbial amylses (pp. 11-27); experiments into, results of and discussion on special problems of the mashing and fermentation process with reference to application of microbial amylases (pp. 28-45); Analytical methods (pp. 46-51); and Resume (pp. 5254).

  7. High quality actively cooled plasma-facing components for fusion

    International Nuclear Information System (INIS)

    Nygren, R.E.

    1995-01-01

    This paper interweaves some suggestions for developing actively cooled plasma-facing components (PFCs) for future fusion devices, with supporting examples taken from the design, fabrication and operation of Tore Supra's Phase III outboard pump limiter (OPL). This actively cooled midplane limiter, designed for heat and particle removal during long-pulse operation, has been operated under essentially thermally steady state conditions. Testing to identify braze flaws, analysis of the impact of joining flaws on the thermal-hydraulic performance of the OPL, and the extensive calorimetry and IR thermography used to confirm and update safe operating limits for power handling of the OPL are reviewed. This experience suggests that, for PFCs in future fusion devices, flaw-tolerant designs are possible; analyses of the impacts of flaws on performance can provide criteria for quality assurance; and validating appropriate methods of inspection for such flaws early in the design development of PFCs is prudent. The need for in-service monitoring is also discussed. (orig.)

  8. Active screen plasma nitriding enhances cell attachment to polymer surfaces

    International Nuclear Information System (INIS)

    Kaklamani, Georgia; Bowen, James; Mehrban, Nazia; Dong, Hanshan; Grover, Liam M.; Stamboulis, Artemis

    2013-01-01

    Active screen plasma nitriding (ASPN) is a well-established technique used for the surface modification of materials, the result of which is often a product with enhanced functional performance. Here we report the modification of the chemical and mechanical properties of ultra-high molecular weight poly(ethylene) (UHMWPE) using 80:20 (v/v) N 2 /H 2 ASPN, followed by growth of 3T3 fibroblasts on the treated and untreated polymer surfaces. ASPN-treated UHMWPE showed extensive fibroblast attachment within 3 h of seeding, whereas fibroblasts did not successfully attach to untreated UHMWPE. Fibroblast-coated surfaces were maintained for up to 28 days, monitoring their metabolic activity and morphology throughout. The chemical properties of the ASPN-treated UHMWPE surface were studied using X-ray photoelectron spectroscopy, revealing the presence of C-N, C=N, and C≡N chemical bonds. The elastic modulus, surface topography, and adhesion properties of the ASPN-treated UHMWPE surface were studied over 28 days during sample storage under ambient conditions and during immersion in two commonly used cell culture media.

  9. Study on the role of active radicals on plasma sterilization inside small diameter flexible polymeric tubes

    Science.gov (United States)

    Mstsuura, Hiroto; Fujiyama, Takatomo; Okuno, Yasuki; Furuta, Masakazu; Okuda, Shuichi; Takemura, Yuichiro

    2015-09-01

    Recently, atmospheric pressure discharge plasma has gathered attention in various fields. Among them, plasma sterilization with many types of plasma source has studied for decades and its mechanism is still an open question. If active radicals produced in plasma has main contribution of killing bacterias, direct contact of the so-called plasma flame might not be necessary. To confirm this, sterilization inside small diameter flexible polymeric tubes is studied in present work. DBD type plasma jet is produce by flowing helium gas in a glass tube. A long polymeric tube is connected and plasma jet is introduced into it. Plasma flame length depends on helium gas flow rate, but limited to about 10 cm in our experimental condition. E.colis set at the exit plasma source is easily killed during 10 min irradiation. At the tube end (about 20 cm away from plasma source exit), sterilization is possible with 30 min operation. This result shows that active radical is produced with helium plasma and mist contained in sample, and it can be transferred more than 20 cm during it life time. More plasma diagnostic data will also be shown at the conference. This work was partially supported by the ''ZE Research Program, IAE(ZE27B-4).

  10. Liver insulinase and insulin-like activity of the blood plasma in irradiated rats

    Energy Technology Data Exchange (ETDEWEB)

    Zhikhareva, A I; Dokshina, G A [Tomskij Gosudarstvennyj Univ. (USSR)

    1975-05-01

    Comparative quantitative analysis of the functional effect of radiation on the activity of liver insulinase of irradiated rats has shown that the insulinase activity of the blood plasma decreases (21-45%) one to three days after the exposure at betatron. Insulinase activity of the liver extracts is also inhibited (16-22%) as compared to intact liver extracts. Twelve days after the exposure and later, insulin-like activity of the plasma and the enzyme activity increase up to 37 per cent.

  11. The enhanced inhibition of water extract of black tea under baking treatment on α-amylase and α-glucosidase.

    Science.gov (United States)

    Tong, Da-Peng; Zhu, Ke-Xue; Guo, Xiao-Na; Peng, Wei; Zhou, Hui-Ming

    2018-02-01

    This paper studied the inhibition of water extract of natural or baked black tea on the activity of α-amylase and α- glucosidase. Baking treatment was found to be one effective way to enhance the inhibition of black tea on both α-amylase and α- glucosidase, and IC 50 of water extract of baked black tea (BBTWE) were 1.213mg/mL and 4.190mg/mL, respectively, while IC 50 of water extract of black tea (BTWE) were 1.723mg/mL and 6.056mg/mL, respectively. This study further studied the mechanism of the effect of water extract on α-amylase and α- glucosidase using HPLC, circular dichroism, and synchronous fluorescence. HPLC analysis of tea polyphenols showed that the content of tea polyphenols with low polarity increased after baking. In addition, BBTWE had higer abilty on decreasing the hydrophobicity of tryptophan residues than BTWE for both α-amylase and α- glucosidase.The increase of α-helix proportion of α-amylase when treated with BBTWE was more obvious than that when treated with BTWE. In a word, thermal process of baked foods may be beneficial for tea polyphenols to reduce the rate of starch digestion. Copyright © 2017 Elsevier B.V. All rights reserved.

  12. A hyper-thermostable α-amylase from Pyrococcus furiosus accumulates in Nicotiana tabacum as functional aggregates.

    Science.gov (United States)

    Zhu, Hong; Reynolds, L Bruce; Menassa, Rima

    2017-06-19

    Alpha amylase hydrolyzes α-bonds of polysaccharides such as starch and produces malto-oligosaccharides. Its starch saccharification applications make it an essential enzyme in the textile, food and brewing industries. Commercially available α-amylase is mostly produced from Bacillus or Aspergillus. A hyper-thermostable and Ca 2++ independent α-amylase from Pyrococcus furiosus (PFA) expressed in E.coli forms insoluble inclusion bodies and thus is not feasible for industrial applications. We expressed PFA in Nicotiana tabacum and found that plant-produced PFA forms functional aggregates with an accumulation level up to 3.4 g/kg FW (fresh weight) in field conditions. The aggregates are functional without requiring refolding and therefore have potential to be applied as homogenized plant tissue without extraction or purification. PFA can also be extracted from plant tissue upon dissolution in a mild reducing buffer containing SDS. Like the enzyme produced in P. furiosus and in E. coli, plant produced PFA preserves hyper-thermophilicity and hyper-thermostability and has a long shelf life when stored in lyophilized leaf tissue. With tobacco's large biomass and high yield, hyper-thermostable α-amylase was produced at a scale of 42 kg per hectare. Tobacco may be a suitable bioreactor for industrial production of active hyperthermostable alpha amylase.

  13. Effect of breed on plasma endothelin-1 concentration, plasma renin activity, and serum cortisol concentration in healthy dogs

    DEFF Research Database (Denmark)

    Höglund, K.; Lequarré, A.-S.; Ljungvall, I.

    2016-01-01

    BACKGROUND: There are breed differences in several blood variables in healthy dogs. OBJECTIVE: Investigate breed variation in plasma endothelin-1 (ET-1) concentration, plasma renin activity, and serum cortisol concentration. ANIMALS: Five-hundred and thirty-one healthy dogs of 9 breeds examined...... at 5 centers (2-4 breeds/center). METHODS: Prospective observational study. Circulating concentrations of ET-1 and cortisol, and renin activity, were measured using commercially available assays. Absence of organ-related or systemic disease was ensured by thorough clinical investigations, including...

  14. Plasma-enhanced atomic layer deposition of silicon dioxide films using plasma-activated triisopropylsilane as a precursor

    International Nuclear Information System (INIS)

    Jeon, Ki-Moon; Shin, Jae-Su; Yun, Ju-Young; Jun Lee, Sang; Kang, Sang-Woo

    2014-01-01

    The plasma-enhanced atomic layer deposition (PEALD) process was developed as a growth technique of SiO 2 thin films using a plasma-activated triisopropylsilane [TIPS, ((iPr) 3 SiH)] precursor. TIPS was activated by an argon plasma at the precursor injection stage of the process. Using the activated TIPS, it was possible to control the growth rate per cycle of the deposited films by adjusting the plasma ignition time. The PEALD technique allowed deposition of SiO 2 films at temperatures as low as 50 °C without carbon impurities. In addition, films obtained with plasma ignition times of 3 s and 10 s had similar values of root-mean-square surface roughness. In order to evaluate the suitability of TIPS as a precursor for low-temperature deposition of SiO 2 films, the vapor pressure of TIPS was measured. The thermal stability and the reactivity of the gas-phase TIPS with respect to water vapor were also investigated by analyzing the intensity changes of the C–H and Si–H peaks in the Fourier-transform infrared spectrum of TIPS

  15. Plasma soluble urokinase plasminogen activator receptor in children with urinary tract infection

    DEFF Research Database (Denmark)

    Wittenhagen, Per; Andersen, Jesper Brandt; Hansen, Anita

    2011-01-01

    In this prospective study we investigated the role of plasma levels of soluble urokinase plasminogen activator receptor (suPAR) in children with urinary tract infection.......In this prospective study we investigated the role of plasma levels of soluble urokinase plasminogen activator receptor (suPAR) in children with urinary tract infection....

  16. Self-consistent dynamo-like activity in turbulent plasmas

    International Nuclear Information System (INIS)

    Bhattacharjee, A.; Hameiri, E.

    1986-05-01

    The evolution of turbulent plasmas is investigated within the framework of resistive magnetohydrodynamics. The functional form of the mean electric field is derived for fluctuations generated by tearing and resistive interchange modes. It is shown that a bath of such local and global modes in pinches causes toroidal field-reversal with finite pressure gradients in the plasma

  17. Antidiabetic and Plasma Endogenous Antioxidant Activity of alstonia ...

    African Journals Online (AJOL)

    Alstonia boonei is a well-known plant of medicinal value but its effect on endogenous plasma antioxidant in diabetes remains unknown. Thus, need to investigate the effects of the methanolic extract of the plant on plasma bilirubin and uric acid level in alloxan induced diabetes rabbits. Twenty five rabbits divided into five ...

  18. Biochemical characterization of digestive carbohydrases from Xanthogaleruca luteola and inhibition of its α-amylase by inhibitors extracted from the common bean

    Directory of Open Access Journals (Sweden)

    Sharifi Mahbobeh

    2011-01-01

    Full Text Available Xanthogaleruca luteola Müll. (Col.: Chrysomelidae is a major urban insect pest on elm trees in Iran. Digestion in the alimentary canal of the elm leaf beetle is facilitated by some carbohydrases which are responsible for the digestion of carbohydrates. The presence of digestive carbohydrases was determined in the digestive system of adult and last larval instar of the elm leaf beetle. The specific activity of α-amylase in the digestive system of adult females and last larval instars were 0.49± 0.05 and 0.72± 0.07 μmol/min/mg protein, respectively. Also, the amylase activity in the midgut of the last larval instar was 3.125- and 4.16-fold higher than that its activity in the foregut and hindgut, respectively. Results showed that optimum activity for α-amylase was found at pH 5. As calculated from Lineweaver-Burk plots, the Km values for α-amylase were 0.64 and 1.44 mg/ml, when glycogen and starch were used as substrates, respectively. The effect of pH and temperature on α- and ß-glucosidase and α- and β-galactosidase activities was determined in the digestive system of X. luteola. Results showed that the activity of α- and ß-glucosidases in adult females was higher than in larvae, but the Яgalactosidase activity in larvae was more than that of the adult. In adult females the glucosidase activity was higher than the galactosidase activity. The zymogram pattern in the native gel revealed that X. luteola α-amylase, β-glucosidase and β-galactosidase in the digestive system had one, three and one isoforms. α-amylase inhibitors, purified from Phaseolus vulgaris L. with an ion-exchange DEAE cellulose column showed good inhibitory activity on X. luteola gut α-amylase.

  19. Evaluation of the effects of a plasma activated medium on cancer cells

    Energy Technology Data Exchange (ETDEWEB)

    Mohades, S.; Laroussi, M., E-mail: mlarouss@odu.edu; Sears, J.; Barekzi, N.; Razavi, H. [Plasma Engineering and Medicine Institute, Old Dominion University, Norfolk, Virginia 23529 (United States)

    2015-12-15

    The interaction of low temperature plasma with liquids is a relevant topic of study to the field of plasma medicine. This is because cells and tissues are normally surrounded or covered by biological fluids. Therefore, the chemistry induced by the plasma in the aqueous state becomes crucial and usually dictates the biological outcomes. This process became even more important after the discovery that plasma activated media can be useful in killing various cancer cell lines. Here, we report on the measurements of concentrations of hydrogen peroxide, a species known to have strong biological effects, produced by application of plasma to a minimum essential culture medium. The activated medium is then used to treat SCaBER cancer cells. Results indicate that the plasma activated medium can kill the cancer cells in a dose dependent manner, retain its killing effect for several hours, and is as effective as apoptosis inducing drugs.

  20. Screening alpha-glucosidase and alpha-amylase inhibitors from natural compounds by molecular docking in silico.

    Science.gov (United States)

    Jhong, Chien-Hung; Riyaphan, Jirawat; Lin, Shih-Hung; Chia, Yi-Chen; Weng, Ching-Feng

    2015-01-01

    The alpha-glucosidase inhibitor is a common oral anti-diabetic drug used for controlling carbohydrates normally converted into simple sugars and absorbed by the intestines. However, some adverse clinical effects have been observed. The present study seeks an alternative drug that can regulate the hyperglycemia by down-regulating alpha-glucosidase and alpha-amylase activity by molecular docking approach to screen the hyperglycemia antagonist against alpha-glucosidase and alpha-amylase activities from the 47 natural compounds. The docking data showed that Curcumin, 16-hydroxy-cleroda-3,13-dine-16,15-olide (16-H), Docosanol, Tetracosanol, Antroquinonol, Berberine, Catechin, Quercetin, Actinodaphnine, and Rutin from 47 natural compounds had binding ability towards alpha-amylase and alpha-glucosidase as well. Curcumin had a better biding ability of alpha-amylase than the other natural compounds. Analyzed alpha-glucosidase activity reveals natural compound inhibitors (below 0.5 mM) are Curcumin, Actinodaphnine, 16-H, Quercetin, Berberine, and Catechin when compared to the commercial drug Acarbose (3 mM). A natural compound with alpha-amylase inhibitors (below 0.5 mM) includes Curcumin, Berberine, Docosanol, 16-H, Actinodaphnine/Tetracosanol, Catechin, and Quercetin when compared to Acarbose (1 mM). When taken together, the implication is that molecular docking is a fast and effective way to screen alpha-glucosidase and alpha-amylase inhibitors as lead compounds of natural sources isolated from medicinal plants. © 2015 International Union of Biochemistry and Molecular Biology.

  1. SusG: A Unique Cell-Membrane-Associated [alpha]-Amylase from a Prominent Human Gut Symbiont Targets Complex Starch Molecules

    Energy Technology Data Exchange (ETDEWEB)

    Koropatkin, Nicole M.; Smith, Thomas J. (Danforth)

    2010-09-21

    SusG is an {alpha}-amylase and part of a large protein complex on the outer surface of the bacterial cell and plays a major role in carbohydrate acquisition by the animal gut microbiota. Presented here, the atomic structure of SusG has an unusual extended, bilobed structure composed of amylase at one end and an unprecedented internal carbohydrate-binding motif at the other. Structural studies further demonstrate that the carbohydrate-binding motif binds maltooligosaccharide distal to, and on the opposite side of, the amylase catalytic site. SusG has an additional starch-binding site on the amylase domain immediately adjacent to the active cleft. Mutagenesis analysis demonstrates that these two additional starch-binding sites appear to play a role in catabolism of insoluble starch. However, elimination of these sites has only a limited effect, suggesting that they may have a more important role in product exchange with other Sus components.

  2. Inducing mechanism of dextrins with different de values on production of alpha-amylase by B. subtilis zjf-1A5

    International Nuclear Information System (INIS)

    Sun, J.; Zhao, R.; Liu, B.

    2014-01-01

    Alpha-amylase was widely used in food industries, textile technology, paper manufacturing and so on. In this paper, the inducing mechanism of corn dextrins with different DE values (dextrose equivalent value) on production of a-amylase by Bacillus subtilis (B.subtilis) ZJF-1A5 was investigated. The results showed that the yield of a-amylase by B.subtilis ZJF-1A5 was increased by using dextrin with a certain DE value range as carbon source, which could be attributed to the presence of oligosaccharide in dextrins. By ordinary fermentation with oligosaccharide as carbon source, it was found that the inducing activity of maltopentaose was the strongest. It could be confirmed that the dextrins played important roles during the process of production of a-amylase by B.subtilis ZJF-1A5. (author)

  3. Study on the immobilization of alpha-amylase by radiation-induced polymerization at low-temperature, (3)

    International Nuclear Information System (INIS)

    Yoshida, Masaru; Kumakura, Minoru; Kaetsu, Isao

    1975-07-01

    The immobilization of α-amylase in high concentration (50-200 mg) by radiation induced polymerization at low temperature, with HEMA has been studied. A feature of the high concentration α-amylase system is phase separation of the mixed solution prior to polymerization, markedly at HEMA concentrations above 50%. Useful immobilization is possible, however, by irradiation of the suspended composition at -196 0 C, which is obtained by shaking the phase-separated system. At temperatures below 0 0 C, the immobilization is possible, but not above this because of the phase separation. The polymerizability of HEMA changes abruptly at 0 0 C. The largest polymerization rate is obtained at -24 0 C, possibly due to phase change by crystallization of water of the buffer solution at 0 0 C. Activity of the immobilized high-concentration α-amylase is as high as 80-85% being somewhat higher than that in the low-concentration case. (auth.)

  4. High-level production of α-amylase by manipulating the expression of alanine racamase in Bacillus licheniformis.

    Science.gov (United States)

    He, Penghui; Zhang, Zeying; Cai, Dongbo; Chen, Yaozhong; Wang, Hao; Wei, Xuetuan; Li, Shunyi; Chen, Shouwen

    2017-09-01

    To improve target protein production by manipulating expression levels of alanine racemase in Bacillus licheniformis. The gene of dal was identified to be responsible for alanine racemase function. Based on the selection marker of dal, a food-grade expression system was constructed in B. licheniformis, and effects of different dal expression levels mediated by promoters on α-amylase production were investigated. The highest α-amylase activity (155 U/ml) was obtained in BL10D/pP43SAT-PtetDal, increased by 27% compared with that of the control strain BL10/pP43SAT in tetracycline-based system (123 U/ml). Moreover, the dal transcriptional level was not correlated positively with that of amyL. A food-grade system for high-level production of α-amylase was constructed in B. licheniformis, revealing that expression levels of selection marker significantly affected target protein production.

  5. Amylase production by solid-state fermentation of agro-industrial wastes using Bacillus sp.

    Directory of Open Access Journals (Sweden)

    Rajshree Saxena

    2011-12-01

    Full Text Available Solid state fermentation was carried out using various agro- industrial wastes with the best amylase producing strain isolated from soil. Different physicochemical conditions were varied for maximum enzyme production. The strain produced about 5400 units/g of amylase at 1:3 moisture content, 20% inoculum, after 72 h of incubation with Mustard Oil seed cake as the substrate. The optimum temperature and pH of the enzyme activity were found to be 50ºC and 6 respectively. The enzyme was found to be thermostable at 70ºC for about 2 h without any salt. It showed stability at pH range 5-7. The metal ions as Na+, Ca++, Mg++ and Co++ enhanced the enzyme activity.

  6. Immobilization of alpha-amylase from Bacillus circulans GRS 313 on coconut fiber.

    Science.gov (United States)

    Dey, Gargi; Nagpal, Varima; Banerjee, Rintu

    2002-01-01

    A simple and inexpensive method for immobilizing alpha-amylase from Bacillus circulans GRS 313 on coconut fiber was developed. The immobilization conditions for highest efficiency were optimized with respect to immobilization pH of 5.5, 30 degrees C, contact time of 4 h, and enzyme to support a ratio of 1:1 containing 0.12 mg/mL of protein. The catalytic properties of the immobilized enzyme were compared with that of the free enzyme. The activity of amylase adsorbed on coconut fiber was 38.7 U/g of fiber at its optimum pH of 5.7 and 48 degrees C, compared with the maximum activity of 40.2 U/mL of free enzyme at the optimum pH of 4.9 and 48 degrees C. The reutilization capacity of the immobilized enzyme was up to three cycles.

  7. Effect of plasma activated water on the postharvest quality of button mushrooms, Agaricus bisporus.

    Science.gov (United States)

    Xu, Yingyin; Tian, Ying; Ma, Ruonan; Liu, Qinghong; Zhang, Jue

    2016-04-15

    Non-thermal plasma is a new approach to improving microbiological safety while maintaining the sensory attributes of the treated foods. Recent research has reported that plasma activated water (PAW) can also efficiently inactivate a wide variety of microorganisms. This study invested the effects of plasma-activated water soaking on the postharvest preservation of button mushrooms (Agaricus bisporus) over seven days of storage at 20°C. Plasma activated water reduced the microbial counts by 1.5 log and 0.5 log for bacteria and fungi during storage, respectively. Furthermore, the corresponding physicochemical and biological properties were assessed between plasma activated water soaking groups and control groups. The results for firmness, respiration rate and relative electrical conductivity suggested that plasma activated water soaking can delay mushroom softening. Meanwhile, no significant change was observed in the color, pH, or antioxidant properties of A. bisporus treated with plasma activated water. Thus, plasma activated water soaking is a promising method for postharvest fresh-keeping of A. bisporus. Copyright © 2015 Elsevier Ltd. All rights reserved.

  8. Properties and Gamma Radiation Stability of Immobilized Alpha Amylase on Synthetic and Natural Polymer Blends

    International Nuclear Information System (INIS)

    Ismaill, S.A.; Mobasher, E.F.; Shousha, M.A.

    2009-01-01

    αAmylase was immobilized onto two different copolymers. One of them was chitosan/alginate copolymer. The other copolymer was N- isopropyl acrylamide and alginate. αAmylase was immobilized by entrapment method. The optimum temperature and thermal inactivation of the free enzyme and the immobilized one were investigated. The activity of the immobilized enzyme was stable at higher temperature. Immobilized enzyme was stable under different ph. The immobilized enzymes showed a slight decrease in the relative activity after being used 12 times. Storage of the free and immobilized enzymes for 2 months showed that the free αamylase lost most of its catalytic activity after storage at this period. The storage of the immobilized enzyme in dry state was much better than that in the wet state. Storage at room temperature showed much less stability of the immobilized enzyme than in 4 degree C. Exposure the free and immobilized enzymes to gamma- radiation at doses (0-50 kGy) showed complete loss in activity of free enzyme at 5 kGy, while the immobilized enzyme showed high resistance to gamma- radiation. The kinetic studies of free and immobilized enzymes showed that the immobilization process increased Km and decreased V m ax values of the enzyme

  9. Properties and Gamma Radiation Stability of Immobilized Alpha Amylase on Synthetic and Natural Polymer Blends

    International Nuclear Information System (INIS)

    Ismaill, S.A.; Mobasher, E.F.; Shousha, M.A.

    2008-01-01

    αAmylase was immobilized onto two different copolymers. One of them was chitosan/alginate copolymer. The other copolymer was N- isopropyl acrylamide and alginate. αAmylase was immobilized by entrapment method. The optimum temperature and thermal inactivation of the free enzyme and the immobilized one were investigated. The activity of the immobilized enzyme was stable at higher temperature. Immobilized enzyme was stable under different ph. The immobilized enzymes showed a slight decrease in the relative activity after being used 12 times. Storage of the free and immobilized enzymes for 2 months showed that the free αamylase lost most of its catalytic activity after storage at this period. The storage of the immobilized enzyme in dry state was much better than that in the wet state. Storage at room temperature showed much less stability of the immobilized enzyme than in 4 degree C. Exposure the free and immobilized enzymes to gamma- radiation at doses (0-50 kGy) showed complete loss in activity of free enzyme at 5 kGy, while the immobilized enzyme showed high resistance to gamma- radiation. The kinetic studies of free and immobilized enzymes showed that the immobilization process increased Km and decreased V m ax values of the enzyme

  10. Study on the immobilization of alpha-amylase by radiation-induced polymerization at low-temperature, (1)

    International Nuclear Information System (INIS)

    Yoshida, Masaru; Kumakura, Minoru; Kaetsu, Isao

    1975-07-01

    The immobilization of enzymes by radiation-induced polymerization at low temperatures has been studied. It is important to know how the enzymes are affected by irradiation. The radiation effect of enzyme itself before immobilization must thus be investigated. In radiation effect on α-amylase from Bacillus subtilis, interesting results were obtained, as follows. The enzyme is very stable for irradiation in the total dose range of 1 x 10 4 to 1 x 10 7 R, and the activity is hardly affected. And further, the relative activity increases by irradiation, when the α-amylase is of high purity or contains some appropriate additive. A certain substance such as diatomaceous earth or CaCl 2 thus decreases the activity, while the addition of DRIERITE composed mainly of CaSO 4 increases the activity. α-Amylase is then more stable and higher in activity in the irradiation at lower temperatures. The activity is independent of presence or absence of the ambient air. In conclusion, α-amylase is very stable for irradiation at low temperatures; therefore, its immobilization by polymerization at low temperature is recommended. (auth.)

  11. Quantitative digital image analysis of chromogenic assays for high throughput screening of alpha-amylase mutant libraries.

    Science.gov (United States)

    Shankar, Manoharan; Priyadharshini, Ramachandran; Gunasekaran, Paramasamy

    2009-08-01

    An image analysis-based method for high throughput screening of an alpha-amylase mutant library using chromogenic assays was developed. Assays were performed in microplates and high resolution images of the assay plates were read using the Virtual Microplate Reader (VMR) script to quantify the concentration of the chromogen. This method is fast and sensitive in quantifying 0.025-0.3 mg starch/ml as well as 0.05-0.75 mg glucose/ml. It was also an effective screening method for improved alpha-amylase activity with a coefficient of variance of 18%.

  12. Low plasma aldosterone despite normal plasma renin activity in uncomplicated type 1 diabetes mellitus : effects of RAAS stimulation

    NARCIS (Netherlands)

    Luik, PT; Kerstens, MN; Hoogenberg, K; Navis, GJ; Dullaart, RPF

    Background Data on levels and responsiveness of PRA and aldosterone in type 1 diabetes mellitus are conflicting. Earlier studies were not standardized with respect to the type of diabetes mellitus, the presence of diabetic complications or sodium intake. Therefore, we studied plasma renin activity

  13. Bactericidal active ingredient in cryopreserved plasma-treated water with the reduced-pH method for plasma disinfection

    Science.gov (United States)

    Kitano, Katsuhisa; Ikawa, Satoshi; Nakashima, Yoichi; Tani, Atsushi; Yokoyama, Takashi; Ohshima, Tomoko

    2016-09-01

    For the plasma disinfection of human body, plasma sterilization in liquid is crucial. We found that the plasma-treated water (PTW) has strong bactericidal activity under low pH condition. Physicochemical properties of PTW is discussed based on chemical kinetics. Lower temperature brings longer half-life and the bactericidal activity of PTW can be kept by cryopreservation. High performance PTW, corresponding to the disinfection power of 22 log reduction (B. subtilis spore), can be obtained by special plasma system equipped with cooling device. This is equivalent to 65% H2O2, 14% sodium hypochlorite and 0.33% peracetic acid, which are deadly poison for human. But, it is deactivated soon at higher temperature (4 sec. at body temperature), and toxicity to human body seems low. For dental application, PTW was effective on infected models of human extracted tooth. Although PTW has many chemical components, respective chemical components in PTW were isolated by ion chromatography. In addition to peaks of H2O2, NO2- and NO3-, a specific peak was detected. and only this fraction had bactericidal activity. Purified active ingredient of PTW is the precursor of HOO, and further details will be discussed in the presentation. MEXT (15H03583, 23340176, 25108505). NCCE (23-A-15).

  14. Molecular Identification of a Newly Isolated Bacillus subtilis BI19 and Optimization of Production Conditions for Enhanced Production of Extracellular Amylase

    Directory of Open Access Journals (Sweden)

    Biplab Kumar Dash

    2015-01-01

    Full Text Available A study was carried out with a newly isolated bacterial strain yielding extracellular amylase. The phylogenetic tree constructed on the basis of 16S rDNA gene sequences revealed this strain as clustered with the closest members of Bacillus sp. and identified as Bacillus subtilis BI19. The effect of various fermentation conditions on amylase production through shake-flask culture was investigated. Rice flour (1.25% as a cheap natural carbon source was found to induce amylase production mostly. A combination of peptone and tryptone as organic and ammonium sulfate as inorganic nitrogen sources gave highest yield. Maximum production was obtained after 24 h of incubation at 37°C with an initial medium pH 8.0. Addition of surfactants like Tween 80 (0.25 g/L and sodium lauryl sulfate (0.2 g/L resulted in 28% and 15% increase in enzyme production, respectively. Amylase production was 3.06 times higher when optimized production conditions were used. Optimum reaction temperature and pH for crude amylase activity were 50°C and 6.0, respectively. The crude enzyme showed activity and stability over a fair range of temperature and pH. These results suggest that B. subtilis BI19 could be exploited for production of amylase at relatively low cost and time.

  15. Molecular Identification of a Newly Isolated Bacillus subtilis BI19 and Optimization of Production Conditions for Enhanced Production of Extracellular Amylase.

    Science.gov (United States)

    Dash, Biplab Kumar; Rahman, M Mizanur; Sarker, Palash Kumar

    2015-01-01

    A study was carried out with a newly isolated bacterial strain yielding extracellular amylase. The phylogenetic tree constructed on the basis of 16S rDNA gene sequences revealed this strain as clustered with the closest members of Bacillus sp. and identified as Bacillus subtilis BI19. The effect of various fermentation conditions on amylase production through shake-flask culture was investigated. Rice flour (1.25%) as a cheap natural carbon source was found to induce amylase production mostly. A combination of peptone and tryptone as organic and ammonium sulfate as inorganic nitrogen sources gave highest yield. Maximum production was obtained after 24 h of incubation at 37 °C with an initial medium pH 8.0. Addition of surfactants like Tween 80 (0.25 g/L) and sodium lauryl sulfate (0.2 g/L) resulted in 28% and 15% increase in enzyme production, respectively. Amylase production was 3.06 times higher when optimized production conditions were used. Optimum reaction temperature and pH for crude amylase activity were 50 °C and 6.0, respectively. The crude enzyme showed activity and stability over a fair range of temperature and pH. These results suggest that B. subtilis BI19 could be exploited for production of amylase at relatively low cost and time.

  16. On methods of measuring the relative plasma composition by active charge exchange

    International Nuclear Information System (INIS)

    Herrmann, W.

    1991-04-01

    Two methods of measuring the hydrogenic composition of plasma with an active diagnostic beam are discussed: Evaluation at equal energies of the neutrals leaving the plasma and at equal velocities. Evaluation at equal velocity has the advantage that the plasma composition, particle penetration and stripping efficiency of the analyzer do not enter the evaluation. The only plasma parameter that has to be known is the plasma temperature. If measurement at two different velocities is possible, the temperature can also be evaluated from the corresponding four fluxes without any further knowledge of plasma or beam parameters. It is discussed under what circumstances evaluation of the plasma composition is possible for non-Maxwellian distribution functions. As the halo effect may be the main source of error, it is shown that the application of a helium beam may considerably reduce this problem. (orig.)

  17. The Importance of Surface-Binding Site towards Starch-Adsorptivity Level in α-Amylase: A Review on Structural Point of View

    Directory of Open Access Journals (Sweden)

    Umi Baroroh

    2017-01-01

    Full Text Available Starch is a polymeric carbohydrate composed of glucose. As a source of energy, starch can be degraded by various amylolytic enzymes, including α-amylase. In a large-scale industry, starch processing cost is still expensive due to the requirement of high temperature during the gelatinization step. Therefore, α-amylase with raw starch digesting ability could decrease the energy cost by avoiding the high gelatinization temperature. It is known that the carbohydrate-binding module (CBM and the surface-binding site (SBS of α-amylase could facilitate the substrate binding to the enzyme’s active site to enhance the starch digestion. These sites are a noncatalytic module, which could interact with a lengthy substrate such as insoluble starch. The major interaction between these sites and the substrate is the CH/pi-stacking interaction with the glucose ring. Several mutation studies on the Halothermothrix orenii, SusG Bacteroides thetaiotamicron, Barley, Aspergillus niger, and Saccharomycopsis fibuligera α-amylases have revealed that the stacking interaction through the aromatic residues at the SBS is essential to the starch adsorption. In this review, the SBS in various α-amylases is also presented. Therefore, based on the structural point of view, SBS is suggested as an essential site in α-amylase to increase its catalytic activity, especially towards the insoluble starch.

  18. Effect of Diuresis on Plasma Renin Activity and Aldosterone Concentration in Normal and Toxemic Pregnancy

    Energy Technology Data Exchange (ETDEWEB)

    Sung, H. K.; Lee, H. S.; Cho, S. S.; Koh, J. H.; Lee, J. K.; Kim, H. S. [Korea Atomic Emergy Research Institute, Seoul (Korea, Republic of)

    1973-03-15

    The changes of plasma renin activity, aldosterone concentration, serum sodium, and potassium levels were studied before and after the water loading followed by diuretics injection. The materials were: 13 non-, 11 normal-, and 11 toxemic pregnancy cases. The plasma renin activity and aldosterone concentration of the cord and postpartum blood were also measured. Following were the results: 1. The plasma renin activity was elevated significantly in normal pregnancy, and slightly in toxemic pregnancy. The serum sodium levels were decreased in pregnancy. 2. The plasma aldosterone concentration was slightly decreased in normal pregnancy, and slightly increased in toxemic pregnancy, however, statistically insignificant. 3. The plasma renin activity of the cord and postpartum blood were lower than those of pregnancy cases. 4. The changes of plasma renin activity after the diuretic administration showed an initial increase, which recovered within 2 hours. These changes were the least in normal pregnancy, and the most in toxemic pregnancy. 5. The changes of plasma aldosterone concentration after the diuretic administration were similar to those of plasma renin activity, although the variations were not so wide.

  19. Effect of Diuresis on Plasma Renin Activity and Aldosterone Concentration in Normal and Toxemic Pregnancy

    International Nuclear Information System (INIS)

    Sung, H. K.; Lee, H. S.; Cho, S. S.; Koh, J. H.; Lee, J. K.; Kim, H. S.

    1973-01-01

    The changes of plasma renin activity, aldosterone concentration, serum sodium, and potassium levels were studied before and after the water loading followed by diuretics injection. The materials were: 13 non-, 11 normal-, and 11 toxemic pregnancy cases. The plasma renin activity and aldosterone concentration of the cord and postpartum blood were also measured. Following were the results: 1. The plasma renin activity was elevated significantly in normal pregnancy, and slightly in toxemic pregnancy. The serum sodium levels were decreased in pregnancy. 2. The plasma aldosterone concentration was slightly decreased in normal pregnancy, and slightly increased in toxemic pregnancy, however, statistically insignificant. 3. The plasma renin activity of the cord and postpartum blood were lower than those of pregnancy cases. 4. The changes of plasma renin activity after the diuretic administration showed an initial increase, which recovered within 2 hours. These changes were the least in normal pregnancy, and the most in toxemic pregnancy. 5. The changes of plasma aldosterone concentration after the diuretic administration were similar to those of plasma renin activity, although the variations were not so wide.

  20. Control plasma renin activity and changes in sympathetic tone as determinants of minoxidil-induced increase in plasma renin activity.

    Science.gov (United States)

    O'Malley, K; Velasco, M; Wells, J; McNay, J L

    1975-01-01

    A study was made of the possible mechanism(s) underlying minoxidil-induced increase in plasma renin activity (PRA). 10 patients with essential hypertension were treated with minoxidil and subsequently with a combination of minoxidil plus propranolol. Minoxidil lowered mean arterial pressure 31.6 plus or minus 3.3 mm Hg, mean plus or minus SEM. There was an associated increase in both PRA, 6.26 plus or minus 2.43 NG/ML/H, and heart rate, 21.4 plus or minus 2.7 beats/min. The changes in PRA and heart rate were positively correlated, r, 0.79. Addition of propranolol reduced mean arterial pressure by a further 10.1 plus or minus 1.5 mm Hg and returned heart rate to control levels. Propranolol reduced PRA significantly but not to control levels. Control PRA positively correlated with PRA on minoxidil, r, 0.97, and with PRA on minoxidil plus propranolol, r, 0.98. We conclude that control PRA is a major determinant of change in PRA with minoxidil. Minoxidil increased PRA by at least two mechanisms: (a) an adrenergic mechanism closely related to change in heart rate and blocked by propranolol, and (b) a mechanism(s) not sensitive to propranolol and possibly related to decrease in renal perfusion pressure. PMID:1127099

  1. Improvement of silicon direct bonding using surfaces activated by hydrogen plasma treatment

    CERN Document Server

    Choi, W B; Lee Jae Sik; Sung, M Y

    2000-01-01

    The plasma surface treatment, using hydrogen gas, of silicon wafers was studied as a pretreatment for silicon direct bonding. Chemical reactions of the hydrogen plasma with the surfaces were used for both surface activation and removal of surface contaminants. Exposure of the silicon wafers to the plasma formed an active oxide layer on the surface. This layer was hydrophilic. The surface roughness and morphology were examined as functions of the plasma exposure time and power. The surface became smoother with shorter plasma exposure time and lower power. In addition, the plasma surface treatment was very efficient in removing the carbon contaminants on the silicon surface. The value of the initial surface energy, as estimated by using the crack propagation method, was 506 mJ/M sup 2 , which was up to about three times higher than the value for the conventional direct bonding method using wet chemical treatments.

  2. Sequential plasma activation methods for hydrophilic direct bonding at sub-200 °C

    Science.gov (United States)

    He, Ran; Yamauchi, Akira; Suga, Tadatomo

    2018-02-01

    We present our newly developed sequential plasma activation methods for hydrophilic direct bonding of silica glasses and thermally grown SiO2 films. N2 plasma was employed to introduce a metastable oxynitride layer on wafer surfaces for the improvement of bond energy. By using either O2-plasma/N2-plasma/N-radical or N2-plasma/N-radical sequential activation, the quartz-quartz bond energy was increased from 2.7 J/m2 to close to the quartz bulk fracture energy that was estimated to be around 9.0 J/m2 after post-bonding annealing at 200 °C. The silicon bulklike bond energy between thermal SiO2 films was also obtained. We suggest that the improvement is attributable to surface modification such as N-related defect formation and asperity softening by the N2 plasma surface treatment.

  3. Kinetics of alpha-amylase secretion in Aspergillus oryzae

    DEFF Research Database (Denmark)

    Henriksen, Anne Laurence Santerre; Carlsen, Morten; Bang de, H.

    1999-01-01

    -chase experiments were carried out to investigate the alpha-amylase secretion kinetics in A. oryzae. No unglycosylated alpha-amylase was detected neither intracellularly nor extracellularly demonstrating that glycosylation was not the rate controlling step in the secretory pathway. The pulse chase experiments...... indicated that there are two pools of intracellular alpha-amylase: a fast secreted and a slow secreted. The secretion of those two pools were described with a kinetic model, which was fitted to the pulse chase experiments. (C) 1999 John Wiley & Sons, Inc. Biotechnol Bioeng 65: 76-82, 1999....

  4. Partial characterization of amylases of two indigenous Central Amazonian rhizobia strains

    Directory of Open Access Journals (Sweden)

    Arlem Nascimento de Oliveira

    2010-02-01

    Full Text Available Amylase production and partial characterization of crude enzyme preparations from two rhizobia strains (R-926 and R-991 were evaluated. For both the strains, maximal amylase activities were achieved during the early-to-mid- exponential growth phase; both were active over a pH range from 4.5 to 8.5 and temperature from 30 to 50 ºC. None of the ions studied (K+, Na+, Ca2+, Hg2+, Mg2+, Mn2+, Cu2+ and Zn2+ was required for the catalytic activity of strain R-926; amylase activity of strain R-991 was stimulated in the presence of K+, Hg2+ and Zn2+. The surfactants SDS, Triton X-100 and Tween-80 did not have a pronounced inhibitory effect on enzyme activities; SDS and Tween-80 caused the highest stimulatory effects. Amylase activities from the rhizobia strains were reduced by up to 30% in the presence of EDTA; amylase activity of R-926 was also inhibited by HgCl2, suggesting that Ca2+and cysteine residues could be important for activity of this strain.A produção e parcial caracterização de extratos brutos de amilase de duas estirpes de rizóbio (R-926 e R-991 foram avaliadas. Para ambas as estirpes, as máximas atividades amilolíticas foram obtidas no início/meio da fase exponencial de crescimento. As amilases rizobiais foram ativas numa variação de pH de 4,5 a 8,5 e temperatura de 30 a 50 ºC. Nenhum dos íons testados (K+, Na+, Ca2+, Hg2+, Mg2+, Mn2+, Cu2+ e Zn2+ foi exigido para a atividade catalítica da estirpe R-926. A amilase produzida pelo R991 foi estimulada na presença de K+, Hg2+ e Zn2+. Os surfactantes SDS, Triton X-100 e Tween-80 não exerceram um pronunciado efeito inibitório sobre as atividades enzimáticas, e SDS e Tween-80 causaram os maiores efeitos estimulatórios. A atividade amilolítica rizobial foi reduzida em até 30% na presença de EDTA; a amilase produzida pela estirpe R-926 também foi inibida pelo HgCl2, sugerindo, ao menos em parte, a importância de Ca2+ e resíduos de cisteína na atividade amilolítica dessa

  5. A Study on Plasma Renin Activity in Korean Hemorrhagic Fever

    International Nuclear Information System (INIS)

    Kim, Suhng Gwon; Cho, Bo Yun; Lee, Jung Sang; Koh, Won Soon; Lee, Mun Ho; Kim, Won Dong; Yun, Hong Jin

    1976-01-01

    To evaluate the possible pathophysiologic role of renin in acute renal failure observed in Korean hemorrhagic fever (KHF), the author measured the basal plasma renin activity (PRA) and the stimulated PRA by radioimmunoassay for angiotensin I in 15 normal controls and 42 KHF patients who are admitted in Seoul National University Hospital and Nation Army Hospital from Jan. 1975 to Jan. 1976. The results obtained were as follows:The mean basal PRA in normal control group was 2.9±2.16 ng/ml/hr in the patients during the oliguric phase of KHF, the mean basal PRA was 4.7±2.13 ng/ml/hr, and there was statistically significant increase compared to the normal control. In the patients during the diuretic phase of KHF, the mean basal PRA was 3.4±2.09 ng/ml/hr, and there was statistically significant decrease compared to the oliguric phase of KHF. In normal control group, the mean basal PRA was 2.9±2.16 ng/ml/hr. And the PRA 1 hour after the administration of Lasix 40 mg intravenously (stimulated PRA) was 5.3±2.20 ng/ml/hr and there was statistically significant increase compared to basal level. In oliguric phase of KHF, the mean basal PRA was 4.6±2.01 ng/ml/hr. And stimulated PRA was 4.4±2.34 ng/ml/hr and there was no significant changes. In diuretic phase of KHF, the mean basal PR was 3.3±1.86 ng/ml/hr. And stimulated PRA was 5.2±2.58 ng/ml/hr and there was statistically significant increase compared to basal level. There were statistically no significant correlations between basal PRA and stimulated PRA and serum creatinine. BUN, urine volume and peritoneal dialysis.

  6. Viscoelastic properties of sweet potato complementary porridges as influenced by endogenous amylases.

    Science.gov (United States)

    Nabubuya, Agnes; Namutebi, Agnes; Byaruhanga, Yusuf; Schuller, Reidar B; Narvhus, Judith; Wicklund, Trude

    2017-11-01

    Sweet potato ( Ipomoea batatas L.) roots contain amylolytic enzymes, which hydrolyze starch thus having the potential to affect the viscosity of sweet potato porridges provided the appropriate working conditions for the enzymes are attained. In this study, the effect of sweet potato variety, postharvest handling conditions, freshly harvested and room/ambient stored roots (3 weeks), and slurry solids content on the viscoelastic properties of complementary porridges prepared using amylase enzyme activation technique were investigated. Five temperatures (55°C, 65°C, 70°C, 75°C, and 80°C) were used to activate sweet potato amylases and the optimum temperature was found to be 75°C. Stored sweet potato roots had higher soluble solids (⁰Brix) content in the pastes compared to fresh roots. In all samples, activation of amylases at 75°C caused changes in the viscoelastic parameters: phase angle (tan δ) and complex viscosity (η * ). Postharvest handling conditions and slurry solids content significantly affected the viscoelastic properties of the porridges with flours from stored roots yielding viscous (liquid-like) porridges and fresh roots producing elastic (solid-like) porridges. Increase in slurry solids content caused reduction in the phase angle values and increase in the viscosity of the sweet potato porridges. The viscosity of the porridges decreased with storage of sweet potato roots. These results provide a possibility for exploiting sweet potato endogenous amylases in the preparation of complementary porridges with both drinkable viscosities and appropriate energy and nutrient densities for children with varying energy needs.

  7. Isolation and characterisation of a novel alpha-amylase from the extreme haloarchaeon Haloterrigena turkmenica.

    Science.gov (United States)

    Santorelli, Marco; Maurelli, Luisa; Pocsfalvi, Gabriella; Fiume, Immacolata; Squillaci, Giuseppe; La Cara, Francesco; Del Monaco, Giovanni; Morana, Alessandra

    2016-11-01

    An extracellular halophilic alpha-amylase (AmyA) was produced by the haloarchaeon Haloterrigena turkmenica grown in medium enriched with 0.2% (w/v) starch. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and size exclusion chromatography (SEC) analyses showed a major band at 66.0kDa and a peak of 54.0kDa, respectively. Analysis of tryptic fragments of the protein present in the major SDS-PAGE band by nano-LC-ESI-MS/MS led to identification of the alpha-amylase catalytic region, encoded by the htur2110 gene, as the protein possessing the described activity. Optimal values for activity were 55°C, pH 8.5 and 2M NaCl, and high thermostability was showed at 55°C and 3M NaCl. AmyA activity was enhanced by Triton X-100 and was not influenced by n-hexane and chloroform. Starch hydrolysis produced different oligomers with maltose as the smallest end-product. The efficiency of AmyA in degrading starch contained in agronomic residues was tested in grape cane chosen as model substrate. Preliminary results showed that starch was degraded making the enzyme a potential candidate for utilization of agro-industrial waste in fuel and chemicals production. AmyA is one of the few investigated amylases produced by haloarchaea, and the first alpha-amylase described among microorganisms belonging to the genus Haloterrigena. Copyright © 2016 Elsevier B.V. All rights reserved.

  8. Adsorption of amylase enzyme on ultrafiltration membranes

    DEFF Research Database (Denmark)

    Beier, Søren; Enevoldsen, Ann Dorrit; Kontogeorgis, Georgios

    2007-01-01

    A method to measure the static adsorption on membrane surfaces has been developed and described. The static adsorption of an amylase-F has been measured on two different ultrafiltration membranes, both with a cut-off value of 10 kDa (a PES membrane and the ETNA10PP membrane, which is a surface......-modified PVDF membrane). The adsorption follows the Langmuir adsorption theory. Thus, the static adsorption consists of monolayer coverage. The static adsorption is expressed both as a permeability drop and an adsorption resistance. From the adsorption isotherms the maximum static permeability drops...... and the maximum static adsorption resistances are determined. The maximum static permeability drop for the hydrophobic PES membrane is 75 % and the maximum static adsorption resistance is 0.014 m2hbar/L. The maximum static permeability drop for the hydrophilic surface-modified PVDF membrane (ETNA10PP) is 23...

  9. Effect of plasma-induced surface charging on catalytic processes: application to CO2 activation

    Science.gov (United States)

    Bal, Kristof M.; Huygh, Stijn; Bogaerts, Annemie; Neyts, Erik C.

    2018-02-01

    Understanding the nature and effect of the multitude of plasma-surface interactions in plasma catalysis is a crucial requirement for further process development and improvement. A particularly intriguing and rather unique property of a plasma-catalytic setup is the ability of the plasma to modify the electronic structure, and hence chemical properties, of the catalyst through charging, i.e. the absorption of excess electrons. In this work, we develop a quantum chemical model based on density functional theory to study excess negative surface charges in a heterogeneous catalyst exposed to a plasma. This method is specifically applied to investigate plasma-catalytic CO2 activation on supported M/Al2O3 (M = Ti, Ni, Cu) single atom catalysts. We find that (1) the presence of a negative surface charge dramatically improves the reductive power of the catalyst, strongly promoting the splitting of CO2 to CO and oxygen, and (2) the relative activity of the investigated transition metals is also changed upon charging, suggesting that controlled surface charging is a powerful additional parameter to tune catalyst activity and selectivity. These results strongly point to plasma-induced surface charging of the catalyst as an important factor contributing to the plasma-catalyst synergistic effects frequently reported for plasma catalysis.

  10. Helium generated cold plasma finely regulates activation of human fibroblast-like primary cells.

    Directory of Open Access Journals (Sweden)

    Paola Brun

    Full Text Available Non-thermal atmospheric pressure plasmas are being developed for a wide range of health care applications, including wound healing. However in order to exploit the potential of plasma for clinical applications, the understanding of the mechanisms involved in plasma-induced activation of fibroblasts, the cells active in the healing process, is mandatory. In this study, the role of helium generated plasma in the tissue repairing process was investigated in cultured human fibroblast-like primary cells, and specifically in hepatic stellate cells and intestinal subepithelial myofibroblasts. Five minutes after treatment, plasma induced formation of reactive oxygen species (ROS in cultured cells, as assessed by flow cytometric analysis of fluorescence-activated 2',7'-dichlorofluorescein diacetate probe. Plasma-induced intracellular ROS were characterized by lower concentrations and shorter half-lives with respect to hydrogen peroxide-induced ROS. Moreover ROS generated by plasma treatment increased the expression of peroxisome proliferator activated receptor (PPAR-γ, nuclear receptor that modulates the inflammatory responses. Plasma exposure promoted wound healing in an in vitro model and induced fibroblast migration and proliferation, as demonstrated, respectively, by trans-well assay and partitioning between daughter cells of carboxyfluorescein diacetate succinimidyl ester fluorescent dye. Plasma-induced fibroblast migration and proliferation were found to be ROS-dependent as cellular incubation with antioxidant agents (e.g. N-acetyl L-cysteine cancelled the biological effects. This study provides evidence that helium generated plasma promotes proliferation and migration in liver and intestinal fibroblast-like primary cells mainly by increasing intracellular ROS levels. Since plasma-evoked ROS are time-restricted and elicit the PPAR-γ anti-inflammatory molecular pathway, this strategy ensures precise regulation of human fibroblast activation and

  11. The impact of respiration and oxidative stress response on recombinant α-amylase production by Saccharomyces cerevisiae

    DEFF Research Database (Denmark)

    Martinez Ruiz, José Luis; Meza, Eugenio; Petranovic, Dina

    2016-01-01

    by overexpressing the endogenous HAP1 gene in a S. cerevisiae strain overproducing recombinant α-amylase. We demonstrate how Hap1p can activate a set of oxidative stress response genes and meanwhile contribute to increase the metabolic rate of the yeast strains, therefore mitigating the negative effect of the ROS...

  12. Concurrent attenuated reactivity of alpha-amylase and cortisol is related to disruptive behavior in male adolescents

    NARCIS (Netherlands)

    Bouw, M.; Jansen, L.M.C.; Vermeiren, R.R.J.M.; Doreleijers, T.A.H.; van de Ven, P.M.; Popma, A.

    2012-01-01

    Attenuated reactivity of salivary alpha-amylase has been proposed as a specific sympathetic marker of disruptive behavior in juveniles and may have additional value to studying other autonomic parameters and hypothalamic-pituitary-adrenal axis activity. Investigating the interrelationships between

  13. Evening salivary alpha-amylase, major depressive disorder, and antidepressant use in the Netherlands Study of Depression and Anxiety (NESDA)

    NARCIS (Netherlands)

    Veen, Gerthe; Giltay, Erik J.; Licht, Carmilla M. M.; Vreeburg, Sophie A.; Cobbaert, Christa M.; Penninx, Brenda W. J. H.; Zitman, Frans G.

    2013-01-01

    Salivary alpha-amylase (sAA) may be a suitable index for sympathetic activity and dysregulation of the autonomic nervous system. The relationship between antidepressants and depression with sAA levels was studied, since antidepressants were previously shown to have a profound impact on heart rate

  14. Overexpression, purification, and characterization of recombinant barley alpha-amylases 1 and 2 secreted by the methylotrophic yeast Pichia pastoris

    DEFF Research Database (Denmark)

    Juge, N; Andersen, Jens S.; Tull, D

    1996-01-01

    , and 2-chloro-4-nitrophenyl beta-D-maltoheptaoside as well as in Ca2+ dependency of activity. Pichia pastoris thus produced in high yields recombinant alpha-amylase that is similar with respect to structure and function to the enzyme purified from malt extracts. This greatly facilitates future mutational...

  15. Heterocyclic Compounds: Effective α-Amylase and α-Glucosidase Inhibitors.

    Science.gov (United States)

    Saeedi, Mina; Hadjiakhondi, Abbas; Nabavi, Seyed Mohammad; Manayi, Azadeh

    2017-01-01

    Diabetes Mellitus (DM) is a metabolic disease characterized by high blood sugar levels. Recently, it has emerged as an important and global health problem with long-term complications and high economic burden. α-Amylase (α-Amy) and α-glucosidase (α-Gls) are two enzymes which are involved in the hydrolysis of starch into sugars and disaccharides leading to the increase of blood glucose level. Hence, inhibition of α-amylase and α-glucosidase plays key role in the treatment of type 2 diabetes. Heterocyclic compounds -both synthetic and naturally occurring derivatives- possess efficient biological properties. At this juncture, they have demonstrated potent inhibitory activity against α-Amy and α-Gls and were found to be versatile tools for the development of novel anti-diabetic agents.

  16. Purification and Characterization of Thermostable and Detergent-Stable α-Amylase from Anoxybacillus sp. AH1

    Directory of Open Access Journals (Sweden)

    Fatma Matpan Bekler

    2016-01-01

    Full Text Available A thermostable and detergent-stable α-amylase from a newly isolated Anoxybacillus sp. AH1 was purifi ed and characterized. Maximum enzyme production (1874.8 U/mL was obtained at 24 h of incubation. The amylase was purified by using Sephadex G-75 gel filtration, after which an 18-fold increase in specific activity and a yield of 9 % were achieved. The molecular mass of the purified enzyme was estimated at 85 kDa by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE. The optimum pH and temperature values of the enzyme were 7.0 and 60 °C, respectively. The enzyme was highly stable in the presence of 30 % glycerol, retaining 85 % of its original activity at 60 °C within 120 min. Km and vmax values were 0.102 μmol and 0.929 μmol/min, espectively, using Lineweaver-Burk plot. The enzyme activity was increased by various detergents, but it was significantly inhibited in the presence of urea. Mg2+ and Ca2+ also significantly activated α-amylase, while Zn2+, Cu2+ and metal ion chelators ethylenediaminetetraacetic acid (EDTA and 1,10-phenanthroline (phen greatly inhibited the enzyme activity. α-Amylase activity was enhanced by β-mercaptoethanol (β-ME and dithiothreitol (DTT to a great extent, but inhibited by p-chloromercuribenzoic acid (PCMB. Iodoacetamide (IAA and N-ethylmaleimide (NEM had a slight, whereas phenylmethylsulfonyl fluoride (PMSF had a strong inhibitory effect on the amylase activity.

  17. Dynamic changes in plasma tissue plasminogen activator, plasminogen activator inhibitor-1 and beta-thromboglobulin content in ischemic stroke.

    Science.gov (United States)

    Zhuang, Ping; Wo, Da; Xu, Zeng-Guang; Wei, Wei; Mao, Hui-ming

    2015-07-01

    The aim of this paper is to investigate the corresponding variations of plasma tissue plasminogen activator (t-PA) and plasminogen activator inhibitor-1 (PAI-1) activities, and beta-thromboglobulin (β-TG) content in patients during different stages of ischemic stroke. Ischemic stroke is a common disease among aging people and its occurrence is associated with abnormalities in the fibrinolytic system and platelet function. However, few reports focus on the dynamic changes in the plasma fibrinolytic system and β-TG content in patients with ischemic stroke. Patients were divided into three groups: acute, convalescent and chronic. Plasma t-PA and PAI-1 activities were determined by chromogenic substrate analysis and plasma β-TG content was detected by radioimmunoassay. Patients in the acute stage of ischemic stroke had significantly increased levels of t-PA activity and β-TG content, but PAI-1 activity was significantly decreased. Negative correlations were found between plasma t-PA and PAI-1 activities and between plasma t-PA activity and β-TG content in patients with acute ischemic stroke. There were significant differences in plasma t-PA and PAI-1 activities in the aged control group, as well as in the acute, convalescent and chronic groups. It can be speculated that the increased activity of t-PA in patients during the acute stage was the result of compensatory function, and that the increase in plasma β-TG level not only implies the presence of ischemic stroke but is likely a cause of ischemic stroke. During the later stages of ischemic stroke, greater attention is required in monitoring levels of PAI-1. Copyright © 2015 Elsevier Ltd. All rights reserved.

  18. Tobacco plants transformed with the bean. alpha. ai gene express an inhibitor of insect. alpha. -amylase in their seeds. [Nicotiana tabacum; Tenebrio molitor

    Energy Technology Data Exchange (ETDEWEB)

    Altabella, T.; Chrispeels, M.J. (Univ. of California, San Diego, La Jolla (USA))

    1990-06-01

    Bean (Phaseolus vulgaris L.) seeds contain a putative plant defense protein that inhibits insect and mammalian but not plant {alpha}-amylases. We recently presented strong circumstantial evidence that this {alpha}-amylase inhibitor ({alpha}Al) is encoded by an already-identified lectin gene whose product is referred to as lectin-like-protein (LLP). We have now made a chimeric gene consisting of the coding sequence of the lectin gene that encodes LLP and the 5{prime} and 3{prime} flanking sequences of the lectin gene that encodes phytohemagglutinin-L. When this chimeric gene was expressed in transgenic tobacco (Nicotiana tabacum), we observed in the seeds a series of polypeptides (M{sub r} 10,000-18,000) that cross-react with antibodies to the bean {alpha}-amylase inhibitor. Most of these polypeptides bind to a pig pancreas {alpha}-amylase affinity column. An extract of the seeds of the transformed tobacco plants inhibits pig pancreas {alpha}-amylase activity as well as the {alpha}-amylase present in the midgut of Tenebrio molitor. We suggest that introduction of this lectin gene (to be called {alpha}ai) into other leguminous plants may be a strategy to protect the seeds from the seed-eating larvae of Coleoptera.

  19. Trypanocidal activity of human plasma on Trypanosoma evansi in mice Atividade tripanocida do plasma humano sobre Trypanosoma evansi em camundongos

    Directory of Open Access Journals (Sweden)

    Aleksandro Schafer Da Silva

    2012-03-01

    Full Text Available This study aimed to test an alternative protocol with human plasma to control Trypanosoma evansi infection in mice. Plasma from an apparently 27-year-old healthy male, blood type A+, was used in the study. A concentration of 100 mg.dL-1 apolipoprotein L1 (APOL1 was detected in the plasma. Forty mice were divided into four groups with 10 animals each. Group A comprised uninfected animals. Mice from groups B, C and D were inoculated with a T. evansi isolate. Group B was used as a positive control. At three days post-infection (DPI, the mice were administered intraperitoneally with human plasma. A single dose of 0.2 mL plasma was given to those in group C. The mice from group D were administered five doses of 0.2 mL plasma with a 24 hours interval between the doses. Group B showed high increasing parasitemia that led to their death within 5 DPI. Both treatments eliminated parasites from the blood and increased the longevity of animals. An efficacy of 50 (group C and 80% (group D of human plasma trypanocidal activity was found using PCR. This therapeutic success was likely achieved in the group D due to their higher levels of APOL1 compared with group C.Este estudo teve como objetivo testar um protocolo alternativo com plasma humano para controlar a infecção por Trypanosoma evansi em camundongos. O plasma foi oriundo de um homem aparentemente saudável, com idade entre 27 anos e tipo de sangue A+. Foi detectada uma concentração de 100 mg.dL -1 de apolipoproteína L1 (APOL1 no plasma. Quarenta camundongos foram divididos em quatro grupos, contendo dez animais cada. Grupo A, composto de animais não infectados. Os roedores dos grupos B, C e D foram inoculados intraperitonealmente com um isolado de T. evansi. O Grupo B foi usado como um controle positivo. Três dias pós-infecção (DPI, os camundongos foram tratados com plasma humano. Uma dose única de 0,2 mL de plasma foi administrada nos roedores do grupo C. Os ratos do grupo D receberam cinco

  20. Cloning and Characterization of an Alpha-amylase Gene from the Hyperthermophilic Archaeon Thermococcus Thioreducens

    Science.gov (United States)

    Bernhardsdotter, Eva C. M. J.; Pusey, Marc L.; Ng, Joseph D.; Garriott, Owen K.

    2004-01-01

    The gene encoding an extracellular a-amylase, TTA, from the hyperthermophilic archaeon Thermococcus thioreducens was cloned and expressed in Escherichia coli. Primary structural analysis revealed high similarity with other a-amylases from the Thermococcus and Pyrococcus genera, as well as the four highly conserved regions typical for a-amylases. The 1374 bp gene encodes a protein of 457 amino acids, of which 435 constitute the mature protein preceded by a 22 amino acid signal peptide. The molecular weight of the purified recombinant enzyme was estimated to be 43 kDa by denaturing gel electrophoresis. Maximal enzymatic activity of recombinant TTA was observed at 90 C and pH 5.5 in the absence of exogenous Ca(2+), and the enzyme was considerably stable even after incubation at 90 C for 2 hours. The thermostability at 90 and 102 C was enhanced in the presence of 5 mM Ca(2+). The extraordinarily high specific activity (about 7.4 x 10(exp 3) U/mg protein at 90 C, pH 5.5 with soluble starch as substrate) together with its low pH optimum makes this enzyme an interesting candidate for starch processing applications.

  1. Kinetics and thermodynamic studies of alpha amylase from bacillus licheniformis mutant

    International Nuclear Information System (INIS)

    Ikram-ul-Haq, M.; Javed, M.M.; Hameed, U.; Adnan, F.

    2010-01-01

    The present investigation deals with the purification and characterization of enzyme a'-amylase from a mutant strain of Bacillus licheniformis EMS-6. A laboratory scale stirred fermentor of 7.5 L capacity was used for the enzyme production under optimal conditions. The enzyme was purified up to homogeneity level by Ammonium sulphate and ion-exchange chromatography using a fast protein liquid chromatography (FPLC) system. The specific activity of the enzyme increased 4-5 times while the yield was found to be 40.4%. The purification fold by RESOURCE-S was recorded to be 3.58. The molecular weight was found to be 55 KDa. In the present research work, the Vmax (2778 U/mg/min) and Km (8.3mg/ml) of a'-amylase were derived from the Lineweaver Burke plot. Thermodynamic parameters for soluble starch hydrolysis, Ea, AH, AS and AG of a'-amylase from B. licheniformis EMS-6 were found to be 25.14 KJ/mol, 22.53 KJ/mole, -110.95 J/mole/K and 36968 J/mole, respectively. The enzyme was stable over a pH range of 4.5-9.0 and gave pH optimum of 7.0. The pKa1 and pKa2 of ionizable groups of active site controlling Vmax, determined by Dixon plot, were 6.0 and 7.5, respectively. (author)

  2. The important role of salivary α-amylase in the gastric digestion of wheat bread starch.

    Science.gov (United States)

    Freitas, Daniela; Le Feunteun, Steven; Panouillé, Maud; Souchon, Isabelle

    2018-01-24

    The role of salivary α-amylase (HSA) in starch digestion is often overlooked in favour of that of pancreatic α-amylase due to the short duration of the oral phase. Although it is generally accepted that the amylase of salivary origin can continue to be active in the stomach, studies ascertaining its contribution are lacking. This study aimed to address this issue by coupling in vitro oral processing with an in vitro dynamic system that mimicked different postprandial gastric pH reduction kinetics observed in vivo following a snack- or lunch-type meal. The digestion of both starch and protein from wheat bread as well as the interplay between the two processes were studied. We have observed that the amylolytic activity of saliva plays a preponderant role hydrolysing up to 80% of bread starch in the first 30 min of gastric digestion. Amylolysis evolved exponentially and nearly superimposing curves were obtained regardless of the acidification profiles, revealing its high efficiency.

  3. Enhanced coagulation activation in preeclampsia: the role of APC resistance, microparticles and other plasma constituents

    NARCIS (Netherlands)

    VanWijk, Marja J.; Boer, Kees; Berckmans, René J.; Meijers, Joost C. M.; van der Post, Joris A. M.; Sturk, Augueste; VanBavel, Ed; Nieuwland, Rienk

    2002-01-01

    Coagulation activation in pregnancy is further enhanced in preeclampsia. We investigated whether this results from increased thrombin generation by the plasma itself or its cell-derived microparticles. Plasma samples were obtained from preeclamptic, normal pregnant and nonpregnant women (each n =

  4. Note: A single-chamber tool for plasma activation and surface functionalization in microfabrication

    Energy Technology Data Exchange (ETDEWEB)

    Bowman, Adam J.; Scherrer, Joseph R.; Reiserer, Ronald S., E-mail: ron.reiserer@vanderbilt.edu [Vanderbilt Institute for Integrative Biosystems Research and Education and Department of Physics and Astronomy, Vanderbilt University, Nashville, Tennessee 37235 (United States)

    2015-06-15

    We present a simple apparatus for improved surface modification of polydimethylsiloxane (PDMS) microfluidic devices. A single treatment chamber for plasma activation and chemical/physical vapor deposition steps minimizes the time-dependent degradation of surface activation that is inherent in multi-chamber techniques. Contamination and deposition irregularities are also minimized by conducting plasma activation and treatment phases in the same vacuum environment. An inductively coupled plasma driver allows for interchangeable treatment chambers. Atomic force microscopy confirms that silane deposition on PDMS gives much better surface quality than standard deposition methods, which yield a higher local roughness and pronounced irregularities in the surface.

  5. Plasma aldosterone concentrations and plasma renin activity in healthy dogs and dogs with hyperadrenocorticism

    NARCIS (Netherlands)

    Javadi, S; Mol, JA; Boer, P; Boer, WH; Runberk, A

    2003-01-01

    The mean (se) basal plasma aldosterone concentrations were significantly lower in 31 dogs with pituitary-dependent hyperadrenocorticism (PDH) (75 [9] pmol/litre) than in 12 healthy dogs (118 [14] pmol/litre), whereas in five dogs with hyperadrenocorticism due to an adrenocortical tumour they were

  6. The influence of α-amylase supplementation, γ-irradiation (60Co) as well as long time of storage of wheat grain on flour technological properties

    International Nuclear Information System (INIS)

    Warchalewski, J.R.

    1989-01-01

    The varieties of winter wheat, Aria and Beta, were studied. The Aria variety was stored for the period of four years. The part of wheat grain from Beta variety was irradiated with γ rays ( 60 Co). In extracts from wheat kernels and flour protein content, total α- and β-amylolytic activity as well as α-amylolytic activity were determined, α-amylases of native and fungal origin were added to the flour obtained from samples of stored wheat kernels (Aria), irradiated and non-irradiated (Beta). Consequently native α-amylase activity of flour increased by 25% and 50%, respectively. Extensive technological estimation of grain and flour with amylase supplements was carried out. The study included: sedimentation analysis, falling number test, milling experiment, farinogram and extensogram analyses, measurement of the degree of damaged starch and flour colour, as well as baking experiment. The obtained experimental loaves of bread were tested for their ability to remain fresh. It was found out that the stored grain flour was characterized by the highest α-amylolytic activity and the lowest falling number value, whereas the irradiated grain flour showed the highest degree of starch damage and water absorption. When α-amylase supplementation to doughs was not accompanied by either irradiation or storage of grain, it definitely changed their physical properties for the worse. The negative influence of native α-amylases appeared to be less significant than that of fungal α-amylases. The positive influence of α-amylase supplementations, especially of those increasing by 25% the native α-amylolytic activity of flour on volume, and freshness of loaves of bread was observed. (author)

  7. Parathyroid mitogenic activity in plasma from patients with familial multiple endocrine neoplasia type 1

    International Nuclear Information System (INIS)

    Brandi, M.L.; Aurbach, G.D.; Fitzpatrick, L.A.; Quarto, R.; Spiegel, A.M.; Bliziotes, M.M.; Norton, J.A.; Doppman, J.L.; Marx, S.J.

    1986-01-01

    Hyperplasia of the parathyroid glands is a central feature of familial multiple endocrine neoplasia type 1. We used cultured bovine parathyroid cells to test for mitogenic activity in plasma from patients with this disorder. Normal plasma stimulated [ 3 H]thymidine incorporation, on the average, to the same extent as it was stimulated in a plasma-free control culture. This contrasted with the results of the tests with plasma from patients with familial multiple endocrine neoplasia type 1, in which parathyroid mitogenic activity increased 2400 percent over the control value (P less than 0.001). Plasma from these patients also stimulated the proliferation of bovine parathyroid cells in culture, whereas plasma from normal subjects inhibited it. Parathyroid mitogenic activity in plasma from the patients with familial multiple endocrine neoplasia type 1 was greater than that in plasma from patients with various other disorders, including sporadic primary hyperparathyroidism (with adenoma, hyperplasia, or cancer of the parathyroid), sporadic primary hypergastrinemia, sporadic pituitary tumor, familial hypocalciuric hypercalcemia, and multiple endocrine neoplasia type 2 (P less than 0.05). Parathyroid mitogenic activity in the plasma of patients with familial multiple endocrine neoplasia type 1 persisted for up to four years after total parathyroidectomy. The plasma also had far more mitogenic activity in cultures of parathyroid cells than did optimal concentrations of known growth factors or of any parathyroid secretagogue. This mitogenic activity had an apparent molecular weight of 50,000 to 55,000. We conclude that primary hyperparathyroidism in familial multiple endocrine neoplasia type 1 may have a humoral cause

  8. Optimization of Thermostable Alpha-Amylase Production Via Mix Agricultural-Residues and Bacillus amyloliquefaciens

    Directory of Open Access Journals (Sweden)

    Shalini RAI

    2014-03-01

    Full Text Available This study reports utilization of mixture of wheat and barley bran (1:1 for the production of thermostable alpha-amylase enzyme through a spore former, heat tolerant strain of Bacillus amyloliquefaciens in solid state fermentation. Maximum yield of alpha-amylase (252.77 U mL-1 was obtained in following optimized conditions, inoculums size 2 mL (2 × 106 CFU/mL, moisture 80%, pH 7±0.02, NaCl (3%, temperature 38±1°C, incubation for 72 h, maltose (1% and tryptone (1%. After SSF crude enzyme was purified via ammonium sulfate precipitation, ion exchange and column chromatography by DEAE Cellulose. Purified protein showed a molecular weight of 42 kDa by SDS-PAGE electrophoresis. After purification, purified enzyme was characterized against several enzymes inhibitors such as temperature, NaCl, pH, metal and surfactants. Pure enzyme was highly active over broad temperature (50-70°C, NaCl concentration (0.5-4 M, and pH (6-10 ranges, indicating it’s a thermoactive and alkali-stable nature. Moreover, CaCl2, MnCl2, =-mercaptoethanol were found to stimulate the amylase activity, whereas FeCl3, sodium dodecyl sulfate (SDS, CuCl3 and ethylenediaminetetraacetic acid (EDTA strongly inhibited the enzyme. Moreover, enzyme specificity and thermal stability conformed by degradation of different soluble starch up to 55°C. Therefore, the present study proved that the extracellular alpha-amylase extracted through wheat flour residues by organism B. amyloliquefaciens MCCB0075, both have considerable potential for industrial application owing to its properties.

  9. Chewing bread: impact on alpha-amylase secretion and oral digestion.

    Science.gov (United States)

    Joubert, Marianne; Septier, Chantal; Brignot, Hélène; Salles, Christian; Panouillé, Maud; Feron, Gilles; Tournier, Carole

    2017-02-22

    During chewing, saliva helps in preparing the food bolus by agglomerating the formed particles, and it initiates enzymatic food breakdown. However, limited information is actually available on the adaptation of saliva composition during the oral processing of complex foods, especially for foods that are sensitive to salivary enzymes. We addressed this question in the context of starch-based products and salivary alpha-amylase. The objectives were two-fold: (1) to determine if salivary alpha-amylase secretion can be modulated by the bread type and (2) to evaluate the contribution of the oral phase in bread enzymatic breakdown. Mouthfuls of three different wheat breads (industrial, artisan and whole-meal breads) were chewed by twelve subjects. Saliva samples were collected at rest and at different times corresponding to 33, 66 and 100% of the individual's chewing sequence. Alpha-amylase activity and total protein content were determined for all saliva samples that were collected. Additionally, the salivary maltose concentration was measured as a marker of bread enzymatic digestion. Boluses were collected at the swallowing time to evaluate the saliva uptake. Chewing industrial bread induced higher saliva uptake than the other breads despite a similar chewing duration. The evolution of salivary amylase activity tended to depend on the type of bread and was highly influenced by a large degree of inter- and intra-subject variability. The protein and maltose concentration steadily increased during chewing as a result of bread breakdown. The salivary protein concentration was mainly affected by the release of the water-soluble proteins of the bread. The salivary maltose concentration was found to be significantly lower for the whole-meal bread. When considering the weight of the mouthful, enzymatic breakdown was found to be most efficient for the breads ranking from industrial > artisan > whole-meal.

  10. Alternative pathways of thromboplastin-dependent activation of human factor X in plasma

    International Nuclear Information System (INIS)

    Marlar, R.A.; Griffin, J.H.

    1981-01-01

    To determine the interrelationships of the major coagulation pathways, the activation of 3H-labeled factor X in normal and various deficient human plasmas was evaluated when clotting was triggered by dilute rabbit or human thromboplastin. Various dilutions of thromboplastin and calcium were added to plasma samples containing 3H-factor X, and the time course of factor X activation was determined. At a 1/250 dilution of rabbit brain thromboplastin, the rate of factor X activation in plasmas deficient in factor VIII or factor IX was 10% of the activation rate of normal plasma or of factor XI deficient plasma. Reconstitution of the deficient plasmas with factors VIII or IX, respectively, reconstituted normal factor X activation. Similar results were obtained when various dilutions of human thromboplastin replaced the rabbit thromboplastin. From these plasma experiments, it is inferred that the dilute thromboplastin-dependent activation of factor X requires factors VII, IX, and VIII. An alternative extrinsic pathway that involves factors IX and VIII may be the physiologic extrinsic pathway and hence help to explain the consistent clinical observations of bleeding diatheses in patients deficient in factors IX or VIII

  11. Comparative Studies on the Amylase and Cellulase Production of ...

    African Journals Online (AJOL)

    Michael Horsfall

    from different sources and examined for their ability to produce cellulase and amylase. Ten fungal ... microorganisms for enzymes and, eventually, also secondary metabolites with anti-microbial or ... polymer i.e. cellulose to smaller sugar.

  12. Amylase production under solid state fermentation by a bacterial ...

    African Journals Online (AJOL)

    SAM

    2014-05-21

    May 21, 2014 ... Addition of exogenous glucose repressed secretion of amylase, demonstrating that a classical glucose effect ... Enzymatic hydrolysis of starch is carried out under tem- ... thermal springs at Arbaminch, Awassa, Nazreth, Shalla.

  13. Thermal stability of α-amylase in aqueous cosolvent systems

    Indian Academy of Sciences (India)

    Prakash

    Department of Protein Chemistry and Technology, Central Food Technological Research ... Keywords. α-Amylase; cosolvents; preferential interaction parameters; thermal stability ...... simulations of trehalose as a 'dynamic reducer' for solvent.

  14. Endogenous pyrogen activity in human plasma after exercise.

    Science.gov (United States)

    Cannon, J G; Kluger, M J

    1983-05-06

    Plasma obtained from human subjects after exercise and injected intraperitoneally into rats elevated rat rectal temperature and depressed plasma iron and zinc concentrations. The pyrogenic component was heat-denaturable and had an apparent molecular weight of 14,000 daltons. Human mononuclear leukocytes obtained after exercise and incubated in vitro released a factor into the medium that also elevated body temperature in rats and reduced trace metal concentrations. These results suggest that endogenous pyrogen, a protein mediator of fever and trace metal metabolism during infection, is released during exercise.

  15. Study of plasma charging-induced white pixel defect increase in CMOS active pixel sensor

    International Nuclear Information System (INIS)

    Tokashiki, Ken; Bai, KeunHee; Baek, KyeHyun; Kim, Yongjin; Min, Gyungjin; Kang, Changjin; Cho, Hanku; Moon, Jootae

    2007-01-01

    Plasma process-induced 'white pixel defect' (WPD) of CMOS active pixel sensor (APS) is studied for Si3N4 spacer etch back process by using a magnetically enhanced reactive ion etching (MERIE) system. WPD preferably takes place at the wafer edge region when the magnetized plasma is applied to Si3N4 etch. Plasma charging analysis reveals that the plasma charge-up characteristic is well matching the edge-intensive WPD generation, rather than the UV radiation. Plasma charging on APS transfer gate might lead to a gate leakage, which could play a role in generation of signal noise or WPD. In this article the WPD generation mechanism will be discussed from plasma charging point of view

  16. Starch Hydrolysis, Polyphenol Contents, and In Vitro Alpha Amylase Inhibitory Properties of Some Nigerian Foods As Affected by Cooking

    Directory of Open Access Journals (Sweden)

    Sani Saidu

    2017-12-01

    Full Text Available The effect of cooking on starch hydrolysis, polyphenol contents, and in vitro α-amylase inhibitory properties of mushrooms (two varieties Russula virescens and Auricularia auricula-judae, sweet potato (Ipomea batatas, and potato (Solanum tuberosum was investigated. The total, resistant, and digestible starch contents of the raw and cooked food samples (FS ranged from 6.4 to 64.9; 0 to 10.1; and 6.4 to 62.7 g/100 g, respectively, while their percentages of starch digestibility (DS values expressed as percentages of total starch hydrolyzed ranged from 45.99 to 100. Raw and boiled unpeeled potato, raw and boiled peeled potato, raw A. auricula-judae, and sweet potato showed mild to high α-amylase inhibition (over a range of concentration of 10–50 mg/mL, which was lower than that of acarbose (that had 69% inhibition of α-amylase over a range of concentration of 2–10 mg/mL, unlike raw R. virescens, boiled A. auricula-judae, and boiled sweet potatoes that activated α-amylase and boiled R. virescens that gave 0% inhibition. The FS contained flavonoids and phenols in addition. The significant negative correlation (r = −0.55; P = 0.05 between the α-amylase inhibitory properties of the raw and cooked FS versus their SD indicates that the α-amylase inhibitors in these FS also influenced the digestibility of their starches. In addition, the significant positive correlation between the α-amylase inhibitory properties of the raw and cooked FS versus their resistant starch (RS (r = 0.59; P = 0.01 contents indicates that the RS constituents of these FS contributed to their α-amylase inhibitory properties. The study showed the usefulness of boiled unpeeled potato, boiled potato peeled, and raw sweet potato as functional foods for people with type 2 diabetes.

  17. 2001 activity report of the development and research line in controlled thermonuclear fusion of the Plasma Associated Laboratory

    International Nuclear Information System (INIS)

    Ludwig, Gerson Otto

    2002-01-01

    The year 2001 activities of the controlled thermonuclear fusion research line of the Plasma Associated Laboratory at the National Institute for Space Research - Brazil are reported. The report approaches the staff, participation in congresses, goals for the year 2002 and papers on Tokamak plasmas, plasma diagnostic, bootstraps, plasma equilibrium and diagnostic

  18. Radiation effects on diamine oxidase activities in intestine and plasma of the rat

    International Nuclear Information System (INIS)

    Ely, M.J.; Speicher, J.M.; Snyder, S.L.; Catravas, G.N.

    1985-01-01

    Diamine oxidase (DAO; EC 1.4.3.6) activity was measured in plasma and ileal tissue homogenates prepared from male Sprague-Dawley rats sacrificed at 1-15 days after acute whole-body irradiation with 14.5-MeV electrons. Animals irradiated with 1 Gy showed no significant changes in plasma and ileal DAO activities through day 13 relative to nonirradiated controls. Animals irradiated with 5, 10 and 12 Gy displayed marked declines in ileal DAO, with levels reaching a nadir on day 3. This was paralleled by a decrease in plasma DAO activity in all three dose groups. Recovery of ileal and plasma DAO levels was later seen as early as day 4 in animals irradiated with 5 and 10 Gy doses, but animals receiving 12 Gy did not survive beyond day 3. A further study highlights the relationship between radiation dose and levels of plasma and mucosal DAO on day 3, the time of maximum decrease at all doses tested. Mucosal DAO activity decreased almost linearly with doses up to 6 Gy. Plasma DAO levels closely paralleled the dose dependency of the mucosal levels. These data suggest that plasma DAO activity might be useful as a readily measurable marker of intestinal epithelial injury and recovery after acute radiation exposure

  19. Active-passive waveguide array for wave excitation in plasmas

    International Nuclear Information System (INIS)

    Motley, R.W.; Hooke, W.M.

    1979-11-01

    A modified version of the standard waveguide grill for exciting lower hybrid plasma waves is proposed. This version should reduce both the number of RF drive components and the amplitude of the (undesirable) surface waves. Results from a simple 2-element array are presented

  20. Plasma dihydroxyphenylalanine (DOPA) is independent of sympathetic activity in humans

    DEFF Research Database (Denmark)

    Eldrup, E; Christensen, N J; Andreasen, J

    1989-01-01

    in diabetic patients with autonomic neuropathy compared to diabetics without neuropathy, whereas baseline plasma DOPA concentrations were similar in the three groups investigated: 6.55 (5.03-7.26, median [interquartile range], n = 8) nmol l-1 in diabetics with neuropathy, 7.41 (5.79-7.97, n = 8) nmol l-1...

  1. Pre-conceptual design activities for the materials plasma exposure experiment

    International Nuclear Information System (INIS)

    Lumsdaine, Arnold; Rapp, Juergen; Varma, Venugopal; Bjorholm, Thomas; Bradley, Craig; Caughman, John; Duckworth, Robert; Goulding, Richard; Graves, Van; Giuliano, Dominic; Lessard, Timothy; McGinnis, Dean; Meitner, Steven

    2016-01-01

    Highlights: • The development of long-pulse nuclear fusion devices requires testing plasma facing components at reactor relevant conditions. • The pre-conceptual design of a proposed linear plasma facility is presented. • Engineering considerations for multiple systems—plasma source and heating, magnet, vacuum, water cooling, and target, are presented. - Abstract: The development of next step fusion facilities such as DEMO or a Fusion Nuclear Science Facility (FNSF) requires first closing technology gaps in some critical areas. Understanding the material-plasma interface is necessary to enable the development of divertors for long-pulse plasma facilities. A pre-conceptual design for a proposed steady-state linear plasma device, the Materials Plasma Exposure Experiment (MPEX), is underway. A helicon plasma source along with ion cyclotron and electron Bernstein wave heating systems will produce ITER divertor relevant plasma conditions with steady-state parallel heat fluxes of up to 40 MW/m"2 with ion fluxes up to 10"2"4/m"2 s on target. Current plans are for the device to use superconducting magnets to produce 1–2 T fields. As a steady-state device, active cooling will be required for components that interact with the plasma (targets, limiters, etc.), as well as for other plasma facing components (transport regions, vacuum tanks, diagnostic ports). Design concepts for the vacuum system, the cooling system, and the plasma heating systems have been completed. The device will include the capability for handling samples that have been neutron irradiated in order to consider the multivariate effects of neutrons, plasma, and high heat-flux on the microstructure of divertor candidate materials. A vacuum cask, which can be disconnected from the high field environment in order to perform in-vacuo diagnosis of the surface evolution is also planned for the facility.

  2. Pre-conceptual design activities for the materials plasma exposure experiment

    Energy Technology Data Exchange (ETDEWEB)

    Lumsdaine, Arnold, E-mail: lumsdainea@ornl.gov; Rapp, Juergen; Varma, Venugopal; Bjorholm, Thomas; Bradley, Craig; Caughman, John; Duckworth, Robert; Goulding, Richard; Graves, Van; Giuliano, Dominic; Lessard, Timothy; McGinnis, Dean; Meitner, Steven

    2016-11-01

    Highlights: • The development of long-pulse nuclear fusion devices requires testing plasma facing components at reactor relevant conditions. • The pre-conceptual design of a proposed linear plasma facility is presented. • Engineering considerations for multiple systems—plasma source and heating, magnet, vacuum, water cooling, and target, are presented. - Abstract: The development of next step fusion facilities such as DEMO or a Fusion Nuclear Science Facility (FNSF) requires first closing technology gaps in some critical areas. Understanding the material-plasma interface is necessary to enable the development of divertors for long-pulse plasma facilities. A pre-conceptual design for a proposed steady-state linear plasma device, the Materials Plasma Exposure Experiment (MPEX), is underway. A helicon plasma source along with ion cyclotron and electron Bernstein wave heating systems will produce ITER divertor relevant plasma conditions with steady-state parallel heat fluxes of up to 40 MW/m{sup 2} with ion fluxes up to 10{sup 24}/m{sup 2} s on target. Current plans are for the device to use superconducting magnets to produce 1–2 T fields. As a steady-state device, active cooling will be required for components that interact with the plasma (targets, limiters, etc.), as well as for other plasma facing components (transport regions, vacuum tanks, diagnostic ports). Design concepts for the vacuum system, the cooling system, and the plasma heating systems have been completed. The device will include the capability for handling samples that have been neutron irradiated in order to consider the multivariate effects of neutrons, plasma, and high heat-flux on the microstructure of divertor candidate materials. A vacuum cask, which can be disconnected from the high field environment in order to perform in-vacuo diagnosis of the surface evolution is also planned for the facility.

  3. High-frequency underwater plasma discharge application in antibacterial activity

    International Nuclear Information System (INIS)

    Ahmed, M. W.; Choi, S.; Lyakhov, K.; Shaislamov, U.; Mongre, R. K.; Jeong, D. K.; Suresh, R.; Lee, H. J.

    2017-01-01

    Plasma discharge is a novel disinfection and effectual inactivation approach to treat microorganisms in aqueous systems. Inactivation of Gram-negative Escherichia coli (E. coli) by generating high-frequency, high-voltage, oxygen (O_2) injected and hydrogen peroxide (H_2O_2) added discharge in water was achieved. The effect of H_2O_2 dose and oxygen injection rate on electrical characteristics of discharge and E. coli disinfection has been reported. Microbial log reduction dependent on H_2O_2 addition with O_2 injection was observed. The time variation of the inactivation efficiency quantified by the log reduction of the initial E. coli population on the basis of optical density measurement was reported. The analysis of emission spectrum recorded after discharge occurrence illustrated the formation of oxidant species (OH"•, H, and O). Interestingly, the results demonstrated that O_2 injected and H_2O_2 added, underwater plasma discharge had fabulous impact on the E. coli sterilization. The oxygen injection notably reduced the voltage needed for generating breakdown in flowing water and escalated the power of discharge pulses. No impact of hydrogen peroxide addition on breakdown voltage was observed. A significant role of oxidant species in bacterial inactivation also has been identified. Furthermore the E. coli survivability in plasma treated water with oxygen injection and hydrogen peroxide addition drastically reduced to zero. The time course study also showed that the retardant effect on E. coli colony multiplication in plasma treated water was favorable, observed after long time. High-frequency underwater plasma discharge based biological applications is technically relevant and would act as baseline data for the development of novel antibacterial processing strategies.

  4. High-frequency underwater plasma discharge application in antibacterial activity

    Energy Technology Data Exchange (ETDEWEB)

    Ahmed, M. W.; Choi, S.; Lyakhov, K.; Shaislamov, U. [Jeju National University, Department of Nuclear and Energy Engineering (Korea, Republic of); Mongre, R. K.; Jeong, D. K. [Jeju National University, Faculty of Biotechnology (Korea, Republic of); Suresh, R.; Lee, H. J., E-mail: hjlee@jejunu.ac.kr [Jeju National University, Department of Nuclear and Energy Engineering (Korea, Republic of)

    2017-03-15

    Plasma discharge is a novel disinfection and effectual inactivation approach to treat microorganisms in aqueous systems. Inactivation of Gram-negative Escherichia coli (E. coli) by generating high-frequency, high-voltage, oxygen (O{sub 2}) injected and hydrogen peroxide (H{sub 2}O{sub 2}) added discharge in water was achieved. The effect of H{sub 2}O{sub 2} dose and oxygen injection rate on electrical characteristics of discharge and E. coli disinfection has been reported. Microbial log reduction dependent on H{sub 2}O{sub 2} addition with O{sub 2} injection was observed. The time variation of the inactivation efficiency quantified by the log reduction of the initial E. coli population on the basis of optical density measurement was reported. The analysis of emission spectrum recorded after discharge occurrence illustrated the formation of oxidant species (OH{sup •}, H, and O). Interestingly, the results demonstrated that O{sub 2} injected and H{sub 2}O{sub 2} added, underwater plasma discharge had fabulous impact on the E. coli sterilization. The oxygen injection notably reduced the voltage needed for generating breakdown in flowing water and escalated the power of discharge pulses. No impact of hydrogen peroxide addition on breakdown voltage was observed. A significant role of oxidant species in bacterial inactivation also has been identified. Furthermore the E. coli survivability in plasma treated water with oxygen injection and hydrogen peroxide addition drastically reduced to zero. The time course study also showed that the retardant effect on E. coli colony multiplication in plasma treated water was favorable, observed after long time. High-frequency underwater plasma discharge based biological applications is technically relevant and would act as baseline data for the development of novel antibacterial processing strategies.

  5. Amylase and chymotrypsinogen synthesis and secretion by the anesthetized rat pancreas

    International Nuclear Information System (INIS)

    Iovanna, J.; Giorgi, D.; Dagorn, J.C.

    1987-01-01

    To investigate the origin of nonparallel secretion, pancreatic juice was collected in the anesthetized rat, during infusion of [ 3 H]phenylalanine. In the basal state, the amylase to chymotrypsinogen ratios of activities and of [ 3 H] incorporations were 3.5 and 2.5 times higher than in the homogenate, respectively. Both ratios decreased to the value in the homogenate upon caerulein stimulation (600 ng/kg). Inhibition of protein synthesis by cycloheximide (15 ng/kg) did not alter enzyme secretion ratios, and depressed basal protein output only partly, suggesting similar secretory pathways for basal and stimulated secretion. Finally, when [ 3 H]phenylalanine was given before anesthesia, the amylase to chymotrypsinogen ratio of incorporations was again higher in basal secretion than in homogenate, even when further protein synthesis was blocked by cycloheximide before urethane injection. Hence, basal secretion comes from a pancreatic compartment which is functional, although minor, in the conscious animal, and shows a higher rate of amylase synthesis, compared to chymotrypsinogen, than the rest of the gland. It could consist of a subpopulation of acinar cells

  6. Application of decolourized and partially purified polygalacturonase and α-amylase in apple juice clarification

    Directory of Open Access Journals (Sweden)

    Tapati Bhanja Dey

    2014-01-01

    Full Text Available Polygalacturonase and α-amylase play vital role in fruit juice industry. In the present study, polygalacturonase was produced by Aspergillus awamori Nakazawa MTCC 6652 utilizing apple pomace and mosambi orange (Citrus sinensis var mosambi peels as solid substrate whereas, α-amylase was produced from A. oryzae (IFO-30103 using wheat bran by solid state fermentation (SSF process. These carbohydrases were decolourized and purified 8.6-fold, 34.8-fold and 3.5-fold, respectively by activated charcoal powder in a single step with 65.1%, 69.8% and 60% recoveries, respectively. Apple juice was clarified by these decolourized and partially purified enzymes. In presence of 1% polygalacturonase from mosambi peels (9.87 U/mL and 0.4% α-amylase (899 U/mL, maximum clarity (%T660nm = 97.0% of juice was attained after 2 h of incubation at 50 ºC in presence of 10 mM CaCl2. Total phenolic content of juice was reduced by 19.8% after clarification, yet with slightly higher %DPPH radical scavenging property.

  7. Application of decolourized and partially purified polygalacturonase and α-amylase in apple juice clarification.

    Science.gov (United States)

    Dey, Tapati Bhanja; Banerjee, Rintu

    2014-01-01

    Polygalacturonase and α-amylase play vital role in fruit juice industry. In the present study, polygalacturonase was produced by Aspergillus awamori Nakazawa MTCC 6652 utilizing apple pomace and mosambi orange (Citrus sinensis var mosambi) peels as solid substrate whereas, α-amylase was produced from A. oryzae (IFO-30103) using wheat bran by solid state fermentation (SSF) process. These carbohydrases were decolourized and purified 8.6-fold, 34.8-fold and 3.5-fold, respectively by activated charcoal powder in a single step with 65.1%, 69.8% and 60% recoveries, respectively. Apple juice was clarified by these decolourized and partially purified enzymes. In presence of 1% polygalacturonase from mosambi peels (9.87 U/mL) and 0.4% α-amylase (899 U/mL), maximum clarity (%T(660 nm) = 97.0%) of juice was attained after 2 h of incubation at 50 °C in presence of 10 mM CaCl2. Total phenolic content of juice was reduced by 19.8% after clarification, yet with slightly higher %DPPH radical scavenging property.

  8. Phenoloxidase activity in larval and juvenile homogenates and adult plasma and haemocytes of bivalve molluscs.

    Science.gov (United States)

    Luna-González, Antonio; Maeda-Martínez, Alfonso N; Vargas-Albores, Francisco; Ascencio-Valle, Felipe; Robles-Mungaray, Miguel

    2003-10-01

    Phenoloxidase (PO) activity was studied in larval and juvenile homogenates and in the plasma and haemocytes of adult Crassostrea gigas, Argopecten ventricosus, Nodipecten subnodosus, and Atrina maura. Samples were tested for the presence of PO activity by incubation with the substrate L-3, 4-dihydroxyphenylalanine using trypsin, alpha-chymotrypsin, laminarin, lipopolysaccharides (LPS), and sodium dodecyl sulphate (SDS) to elicit activation of prophenoloxidase (proPO) system. PO activity was not detected in larval homogenate. In juvenile homogenate, PO activity was found only in C. gigas and N. subnodosus. PO activity was present in adult samples and was enhanced by elicitors in the plasma of all species tested, but in haemocyte lysate supernatant (HLS) of only N. subnodosus. Activation of proPO by laminarin was suppressed by a protease inhibitor cocktail (P-2714) in plasma and HLS of all species tested.

  9. Relationship between plasma xanthine oxidoreductase activity and left ventricular ejection fraction and hypertrophy among cardiac patients.

    Directory of Open Access Journals (Sweden)

    Yuki Fujimura

    Full Text Available Xanthine oxidoreductase (XOR, which catalyzes purine catabolism, has two interconvertible forms, xanthine dehydrogenase and xanthine oxidase, the latter of which produces superoxide during uric acid (UA synthesis. An association between plasma XOR activity and cardiovascular and renal outcomes has been previously suggested. We investigated the potential association between cardiac parameters and plasma XOR activity among cardiology patients.Plasma XOR activity was measured by [13C2,15N2]xanthine coupled with liquid chromatography/triplequadrupole mass spectrometry. Among 270 patients who were not taking UA-lowering drugs, XOR activity was associated with body mass index (BMI, alanine aminotransferase (ALT, HbA1c and renal function. Although XOR activity was not associated with serum UA overall, patients with chronic kidney disease (CKD, those with higher XOR activity had higher serum UA among patients without CKD. Compared with patients with the lowest XOR activity quartile, those with higher three XOR activity quartiles more frequently had left ventricular hypertrophy. In addition, plasma XOR activity showed a U-shaped association with low left ventricular ejection fraction (LVEF and increased plasma B-type natriuretic peptide (BNP levels, and these associations were independent of age, gender, BMI, ALT, HbA1C, serum UA, and CKD stages.Among cardiac patients, left ventricular hypertrophy, low LVEF, and increased BNP were significantly associated with plasma XOR activity independent of various confounding factors. Whether pharmaceutical modification of plasma XOR activity might inhibit cardiac remodeling and improve cardiovascular outcome should be investigated in future studies.

  10. The effects of residual platelets in plasma on plasminogen activator inhibitor-1 and plasminogen activator inhibitor-1-related assays

    NARCIS (Netherlands)

    M. Pieters (Marlien); S.A. Barnard (Sunelle A.); D.T. Loots (Du Toit); D.C. Rijken (Dingeman)

    2017-01-01

    textabstractDue to controversial evidence in the literature pertaining to the activity of plasminogen activator inhibitor-1 in platelets, we examined the effects of residual platelets present in plasma (a potential pre-analytical variable) on various plasminogen activator inhibitor-1 and plasminogen

  11. The Effect of a Brief Salivary α-Amylase Exposure During Chewing on Subsequent in Vitro Starch Digestion Curve Profiles

    Directory of Open Access Journals (Sweden)

    Charles S. Brennan

    2010-07-01

    Full Text Available There is inconsistency between current in vitro digestion methods with regard to accommodation of a (salivary α-amylase exposure during the oral phase. The effect of a salivary α-amylase pre-exposure on subsequent in vitro starch digestion curve profiles for various foods was investigated. Foods were chewed, expectorated and the boluses left to rest for 0–15 min. During pancreatic digestion, aliquots were taken and hydrolysis curves constructed for comparison against those of the same foods comminuted with a manually-operated chopper, hence spared exposure to saliva. Hydrolysate aliquots taken at T0 (time zero of the digestion of chewed samples contained higher levels of glucose and dextrins compared with chopped samples. Pancreatin activity immediately overwhelmed differences in sugar released due to salivary amylase activity. Within 10 min no differences were detectable between hydrolysis curves for chewed and chopped foods. Salivary amylase pretreatment does not contribute to the robustness or relative accuracy of in vitro methods.

  12. Production of α-amylase from Streptomyces sp. SLBA-08 strain using agro-industrial by-products

    Directory of Open Access Journals (Sweden)

    Édilla Ribeiro dos Santos

    2012-10-01

    Full Text Available Approximately 1.5 trillion tons are the estimated yearly biomass production, making it an essentially unlimited source of raw material for environmentally friendly and biocompatible products transformed by microorganism, specially fungi and actinomycetes. Several lignocellulosic residues, such as sisal waste and sugarcane bagasse contain starch in their structures which could become important sources for the production of amylases. This study evaluated the production of amylolytic enzymes using Streptomyces sp. SLBA-08 strain, isolated from a semi-arid soil, according to their ability to grow on soluble starch as the sole carbon source. The effect of the carbon source (sisal waste and sugarcane bagasse on α-amylase production was studied using submerged cultivations at 30 ºC. The highest level of α-amylase activity corresponded to 10.1 U. mL-1 and was obtained using sisal waste (2.7% and urea (0.8% in submerged fermentation after 3 days of cultivation. The partial characterization showed the best α-amylase activity at 50ºC and pH 7.0. These results are of great importance for the use of sisal waste as a substrate for biotechnological proposes.

  13. Active control of magneto-hydrodynamic instabilities in hot plasmas

    CERN Document Server

    2015-01-01

    During the past century, world-wide energy consumption has risen dramatically, which leads to a quest for new energy sources. Fusion of hydrogen atoms in hot plasmas is an attractive approach to solve the energy problem, with abundant fuel, inherent safety and no long-lived radioactivity.  However, one of the limits on plasma performance is due to the various classes of magneto-hydrodynamic instabilities that may occur. The physics and control of these instabilities in modern magnetic confinement fusion devices is the subject of this book. Written by foremost experts, the contributions will provide valuable reference and up-to-date research reviews for "old hands" and newcomers alike.

  14. Atmospheric-pressure plasma activation and surface characterization on polyethylene membrane separator

    Science.gov (United States)

    Tseng, Yu-Chien; Li, Hsiao-Ling; Huang, Chun

    2017-01-01

    The surface hydrophilic activation of a polyethylene membrane separator was achieved using an atmospheric-pressure plasma jet. The surface of the atmospheric-pressure-plasma-treated membrane separator was found to be highly hydrophilic realized by adjusting the plasma power input. The variations in membrane separator chemical structure were confirmed by Fourier transform infrared spectroscopy and X-ray photoelectron spectroscopy. Chemical analysis showed newly formed carbonyl-containing groups and high surface concentrations of oxygen-containing species on the atmospheric-pressure-plasma-treated polymeric separator surface. It also showed that surface hydrophilicity primarily increased from the polar component after atmospheric-pressure plasma treatment. The surface and pore structures of the polyethylene membrane separator were examined by scanning electron microscopy, revealing a slight alteration in the pore structure. As a result of the incorporation of polar functionalities by atmospheric-pressure plasma activation, the electrolyte uptake and electrochemical impedance of the atmospheric-pressure-plasma-treated membrane separator improved. The investigational results show that the separator surface can be controlled by atmospheric-pressure plasma surface treatment to tailor the hydrophilicity and enhance the electrochemical performance of lithium ion batteries.

  15. G-protein activity in Percoll-purified plasma membranes, bulk plasma membranes, and low-density plasma membranes isolated from rat cerebral cortex

    Czech Academy of Sciences Publication Activity Database

    Bouřová, Lenka; Stöhr, Jiří; Lisý, Václav; Rudajev, Vladimír; Novotný, Jiří; Svoboda, Petr

    2009-01-01

    Roč. 15, č. 4 (2009), BR111-BR122 ISSN 1234-1010 R&D Projects: GA MŠk(CZ) LC554; GA MŠk(CZ) LC06063; GA ČR(CZ) GA309/06/0121; GA AV ČR(CZ) IAA500110606 Institutional research plan: CEZ:AV0Z50110509 Keywords : rat cerebral cortex * plasma membrane * G-protein activity Subject RIV: CE - Biochemistry Impact factor: 1.543, year: 2009

  16. The α-Amylase Induction in Endosperm during Rice Seed Germination Is Caused by Gibberellin Synthesized in Epithelium1

    Science.gov (United States)

    Kaneko, Miyuki; Itoh, Hironori; Ueguchi-Tanaka, Miyako; Ashikari, Motoyuki; Matsuoka, Makoto

    2002-01-01

    We recently isolated two genes (OsGA3ox1 and OsGA3ox2) from rice (Oryza sativa) encoding 3β-hydroxylase, which catalyzes the final step of active gibberellin (GA) biosynthesis (H. Itoh, M. Ueguchi-Tanaka, N. Sentoku, H. Kitano, M. Matsuoka, M. Kobayashi [2001] Proc Natl Acad Sci USA 98: 8909–8914). Using these cloned cDNAs, we analyzed the temporal and spatial expression patterns of the 3β-hydroxylase genes and also an α-amylase gene (RAmy1A) during rice seed germination to investigate the relationship between GA biosynthesis and α-amylase expression. Northern-blot analyses revealed that RAmy1A expression in the embryo occurs before the induction of 3β-hydroxylase expression, whereas in the endosperm, a high level of RAmy1A expression occurs 1 to 2 d after the peak of OsGA3ox2 expression and only in the absence of uniconazol. Based on the analysis of an OsGA3ox2 null mutant (d18-Akibare dwarf), we determined that 3β-hydroxylase produced by OsGA3ox2 is important for the induction of RAmy1A expression and that the OsGA3ox1 product is not essential for α-amylase induction. The expression of OsGA3ox2 was localized to the shoot region and epithelium of the embryo, strongly suggesting that active GA biosynthesis occurs in these two regions. The synthesis of active GA in the epithelium is important for α-amylase expression in the endosperm, because an embryonic mutant defective in shoot formation, but which developed epithelium cells, induced α-amylase expression in the endosperm, whereas a mutant defective in epithelium development did not. PMID:11950975

  17. Active calcium transport in plasma membrane vesicles from developing cotyledons of common bean

    International Nuclear Information System (INIS)

    Huang Jianzhong; Chen Ziyuan

    1995-01-01

    Plasma membrane vesicles were prepared from the developing cotyledons of common bean (Phaseolus vulgaris L cv Diyundou) by aqueous two-phase partitioning and characterized as to their purity by assaying marker enzymes for other membranes. The putative plasma membrane fraction was minimally contaminated by membranes other than plasma membrane and hence was of high purity. It exhibited a Ca 2+ -dependent ATPase activity, which was inhibited by 1 μmol/L EB and promoted by calcium ionophore A23187. Such an activity was responsible for the observed ATP-dependent 45 Ca 2+ uptake into inside-out plasma membrane vesicles. This process was stimulated by 0.6 μmol/L CaM and 20 μmol/L IAA but inhibited by 2 μmol/L ABA and abolished by A23187. Possible role of cytoplasmic Ca 2+ in mediating phytohormones activity is discussed

  18. Enhanced oxidation of naphthalene using plasma activation of TiO2/diatomite catalyst.

    Science.gov (United States)

    Wu, Zuliang; Zhu, Zhoubin; Hao, Xiaodong; Zhou, Weili; Han, Jingyi; Tang, Xiujuan; Yao, Shuiliang; Zhang, Xuming

    2018-04-05

    Non-thermal plasma technology has great potential in reducing polycyclic aromatic hydrocarbons (PAHs) emission. But in plasma-alone process, various undesired by-products are produced, which causes secondary pollutions. Here, a dielectric barrier discharge (DBD) reactor has been developed for the oxidation of naphthalene over a TiO 2 /diatomite catalyst at low temperature. In comparison to plasma-alone process, the combination of plasma and TiO 2 /diatomite catalyst significantly enhanced naphthalene conversion (up to 40%) and CO x selectivity (up to 92%), and substantially reduced the formation of aerosol (up to 90%) and secondary volatile organic compounds (up to near 100%). The mechanistic study suggested that the presence of the TiO 2 /diatomite catalyst intensified the electron energy in the DBD. Meantime, the energized electrons generated in the discharge activated TiO 2 , while the presence of ozone enhanced the activity of the TiO 2 /diatomite catalyst. This plasma-catalyst interaction led to the synergetic effect resulting from the combination of plasma and TiO 2 /diatomite catalyst, consequently enhanced the oxidation of naphthalene. Importantly, we have demonstrated the effectiveness of plasma to activate the photocatalyst for the deep oxidation of PAH without external heating, which is potentially valuable in the development of cost-effective gas cleaning process for the removal of PAHs in vehicle applications during cold start conditions. Copyright © 2017 Elsevier B.V. All rights reserved.

  19. Plasma Membrane ATPase Activity following Reversible and Irreversible Freezing Injury 1

    Science.gov (United States)

    Iswari, S.; Palta, Jiwan P.

    1989-01-01

    Plasma membrane ATPase has been proposed as a site of functional alteration during early stages of freezing injury. To test this, plasma membrane was purified from Solanum leaflets by a single step partitioning of microsomes in a dextran-polyethylene glycol two phase system. Addition of lysolecithin in the ATPase assay produced up to 10-fold increase in ATPase activity. ATPase activity was specific for ATP with a Km around 0.4 millimolar. Presence of the ATPase enzyme was identified by immunoblotting with oat ATPase antibodies. Using the phase partitioning method, plasma membrane was isolated from Solanum commersonii leaflets which had four different degrees of freezing damage, namely, slight (reversible), partial (partially reversible), substantial and total (irreversible). With slight (reversible) damage the plasma membrane ATPase specific activity increased 1.5- to 2-fold and its Km was decreased by about 3-fold, whereas the specific activity of cytochrome c reductase and cytochrome c oxidase in the microsomes were not different from the control. However, with substantial (lethal, irreversible) damage, there was a loss of membrane protein, decrease in plasma membrane ATPase specific activity and decrease in Km, while cytochrome c oxidase and cytochrome c reductase were unaffected. These results support the hypothesis that plasma membrane ATPase is altered by slight freeze-thaw stress. Images Figure 1 Figure 2 PMID:16666856

  20. Formation of tissue factor activity following incubation of recombinant human tissue factor apoprotein with plasma lipoproteins

    International Nuclear Information System (INIS)

    Sakai, T.; Kisiel, W.

    1990-01-01

    Incubation of recombinant human tissue factor apoprotein (Apo-TF) with human plasma decreased the recalcified clotting time of this plasma in a time-and dose-dependent manner suggesting relipidation of the Apo-TF by plasma lipoproteins. Incubation of Apo-TF with purified preparations of human very low density, low density and high density lipoproteins resulted in tissue factor activity in a clotting assay. The order of effectiveness was VLDL greater than LDL much greater than HDL. Tissue factor activity generated by incubation of a fixed amount of Apo-TF with plasma lipoproteins was lipoprotein concentration-dependent and saturable. The association of Apo-TF with lipoprotein particles was supported by gel filtration studies in which 125 I-Apo-TF coeluted with the plasma lipoprotein in the void volume of a Superose 6 column in the presence and absence of calcium ions. In addition, void-volume Apo-TF-lipoprotein fractions exhibited tissue factor activity. These results suggest that the factor VIII-bypassing activity of bovine Apo-TF observed in a canine hemophilic model may be due, in part, to its association with plasma lipoproteins and expression of functional tissue factor activity

  1. Effect of γ irradiation on the activity of lecithin-cholesterol acyltransferase in plasma of the rat

    International Nuclear Information System (INIS)

    Dousset, N.; Douste-Blazy, L.

    1975-01-01

    Plasma cholesterol and lecithin-cholesterol-acyl-transferase activity are studied in irradiated rats. Ionizing radiations cause an increase of cholesterol levels in plasma, concerning mainly ester fraction. Lecithin-cholesterol-acyltransferase activity in plasma of irradiated rats is lowered 48 hours after exposure. This decreased rate of LCAT is probably the consequence of the post-irradiation hypercholesterolemia [fr

  2. [Are amylases in bakery products and flour potential food allergens?].

    Science.gov (United States)

    Baur, X; Sander, I; Jansen, A; Czuppon, A B

    1994-05-21

    The enzyme alpha-amylase from the mould Aspergillus oryzae (Asp o II) routinely used for the production of bread, cakes and pastries has in recent years been identified as an inhalative allergen for occupational diseases (bakers' asthma). It is doubtful whether this amylase in the final product, i.e. after the baking procedure, can still be regarded as an allergen. To clarify this question, detailed case histories on 138 subjects were recorded (98 allergics, 20 patients suffering form chronic intestinal diseases, 20 healthy controls). The clinical examinations included prick skin test and IgE antibody determination using one of the customary enzyme preparations. EAST showed a few of these 138 bread consumers to be weakly sensitized to the enzyme. One of the subjects displayed a significant reaction to alpha-amylase heated to 200 degrees C. As expected, eleven bakers sensitized to alpha-amylase by inhaling it in the workplace (positive prick test, positive case history) predominantly exhibited specific IgE antibodies to the native enzyme. Apart from one weakly positive finding, heated alpha-amylase yielded negative results in this collective. Baking conditions vary widely, especially with regard to single components, temperature and duration. Thus, further investigations as to residual allergenicity or the feasible occurrence of new antigenic determinants during the production of bread, cake and pastries are required. 27% of bakers examined and 9% of atopics showed antibodies to a flour inherent enzyme, a beta-amylase. On the whole, the selected conditions hinted at a weakly sensitizing potential inherent in baking flour and in added amylase.

  3. Lipase or amylase for the diagnosis of acute pancreatitis?

    Science.gov (United States)

    Ismail, Ola Z; Bhayana, Vipin

    2017-12-01

    Acute pancreatitis is a rapid onset of inflammation of the pancreas causing mild to severe life threatening conditions [1, 2]. In Canada, acute pancreatitis is the 5th most expensive digestive disease in Canada with a considerable economic burden on the health care system [3]. The diagnosis of acute pancreatitis is usually based on the presence of abdominal pain and elevated levels of serum amylase and/or lipase. Many health care centers use either serum amylase, lipase or both to diagnose acute pancreatitis without considering which one could provide a better diagnostic accuracy. The aim of this review is to investigate whether serum lipase alone is a sufficient biomarker for the diagnosis of acute pancreatitis. We have examined various studies looking at the utilization, sensitivity, specificity and cost associated savings of lipase and amylase in the diagnosis of acute pancreatitis. When comparing different studies, serum lipase offers a higher sensitivity than serum amylase in diagnosing acute pancreatitis. Lipase also offers a larger diagnostic window than amylase since it is elevated for a longer time, thus allowing it to be a useful diagnostic biomarker in early and late stages of acute pancreatitis. Several recent evidence-based guidelines recommend the use of lipase over amylase. Nevertheless, both lipase and amylase alone lack the ability to determine the severity and etiology of acute pancreatitis. The co-ordering of both tests has shown little to no increase in the diagnostic sensitivity and specificity. Thus, unnecessary testing and laboratory expenditures can be reduced by testing lipase alone. Copyright © 2017 The Canadian Society of Clinical Chemists. Published by Elsevier Inc. All rights reserved.

  4. Effect of platelet activating factor on endothelial permeability to plasma macromolecules

    International Nuclear Information System (INIS)

    Handley, D.A.; Arbeeny, C.M.; Lee, M.L.; Van Valen, R.G.; Saunders, R.N.

    1984-01-01

    The effect of intrajugular administration of platelet activating factor (PAF-C16) on vascular permeability was examined in the guinea pig. To examine the loss of selective endothelial permeability, the extravasative effect of PAF was assessed by monitoring hemoconcentration and the plasma loss of 125 I-albumin (6.7 nm), 125 I-low density lipoproteins (22.0 nm) or 125 I-very low density lipoproteins (62.1 nm). Extravasation was dose-dependent and began 1 min after PAF administration, continuing for 5-7 min. During extravasation, there was no evidence for selective plasma retention of any of the labeled plasma tracers, as measured by plasma radioactivity. These results suggest that PAF-induced extravasation is dose-dependent, with increases in vascular permeability sufficient to permit similar plasma efflux rates of albumin, low density lipoproteins and very low density lipoproteins

  5. Plasma-activated core-shell gold nanoparticle films with enhanced catalytic properties

    Energy Technology Data Exchange (ETDEWEB)

    Llorca, Jordi, E-mail: jordi.llorca@upc.edu; Casanovas, Albert; Dominguez, Montserrat; Casanova, Ignasi [Universitat Politecnica de Catalunya, Institut de Tecniques Energetiques (Spain); Angurell, Inmaculada; Seco, Miquel; Rossell, Oriol [Universitat de Barcelona, Departament de Quimica Inorganica (Spain)

    2008-03-15

    Catalytically active gold nanoparticle films have been prepared from core-shell nanoparticles by plasma-activation and characterized by high-resolution transmission electron microscopy, atomic force microscopy, and X-ray photoelectron spectroscopy. Methane can be selectively oxidized into formic acid with an O{sub 2}-H{sub 2} mixture in a catalytic wall reactor functionalized with plasma-activated gold nanoparticle films containing well-defined Au particles of about 3.5 nm in diameter. No catalytic activity was recorded over gold nanoparticle films prepared by thermal decomposition of core-shell nanoparticles due to particle agglomeration.

  6. Plasma-activated core-shell gold nanoparticle films with enhanced catalytic properties

    International Nuclear Information System (INIS)

    Llorca, Jordi; Casanovas, Albert; Dominguez, Montserrat; Casanova, Ignasi; Angurell, Inmaculada; Seco, Miquel; Rossell, Oriol

    2008-01-01

    Catalytically active gold nanoparticle films have been prepared from core-shell nanoparticles by plasma-activation and characterized by high-resolution transmission electron microscopy, atomic force microscopy, and X-ray photoelectron spectroscopy. Methane can be selectively oxidized into formic acid with an O 2 -H 2 mixture in a catalytic wall reactor functionalized with plasma-activated gold nanoparticle films containing well-defined Au particles of about 3.5 nm in diameter. No catalytic activity was recorded over gold nanoparticle films prepared by thermal decomposition of core-shell nanoparticles due to particle agglomeration

  7. Codon Optimization Significantly Improves the Expression Level of α-Amylase Gene from Bacillus licheniformis in Pichia pastoris

    Directory of Open Access Journals (Sweden)

    Jian-Rong Wang

    2015-01-01

    Full Text Available α-Amylase as an important industrial enzyme has been widely used in starch processing, detergent, and paper industries. To improve expression efficiency of recombinant α-amylase from Bacillus licheniformis (B. licheniformis, the α-amylase gene from B. licheniformis was optimized according to the codon usage of Pichia pastoris (P. pastoris and expressed in P. pastoris. Totally, the codons encoding 305 amino acids were optimized in which a total of 328 nucleotides were changed and the G+C content was increased from 47.6 to 49.2%. The recombinants were cultured in 96-deep-well microplates and screened by a new plate assay method. Compared with the wild-type gene, the optimized gene is expressed at a significantly higher level in P. pastoris after methanol induction for 168 h in 5- and 50-L bioreactor with the maximum activity of 8100 and 11000 U/mL, which was 2.31- and 2.62-fold higher than that by wild-type gene. The improved expression level makes the enzyme a good candidate for α-amylase production in industrial use.

  8. Alpha-Amylase Inhibition and Antioxidative Capacity of Some Antidiabetic Plants Used by the Traditional Healers in Southeastern Nigeria

    Directory of Open Access Journals (Sweden)

    Sunday O. Oyedemi

    2017-01-01

    Full Text Available Oxidative stress plays a significant role in the pathogenesis of metabolic syndrome including diabetes mellitus (DM. The inhibition of alpha-amylase is an important therapeutic target in the regulation of postprandial increase of blood glucose in diabetic patients. The present study investigated the alpha-amylase inhibitory and antioxidant potential of selected herbal drugs used in the treatment of DM by the traditional healers in Isiala Mbano and Ikwuano regions of southeastern Nigeria. Antioxidant activity was evaluated in terms of free radical scavenging, reducing power, and total phenolic (TPC and flavonoid content (TFC in consonance with the TLC profiling. The results showed that methanol crude extracts from Anacardium occidentale (AO and Ceiba pentandra (CP recorded higher TPC and TFC, potent free radical scavenging, and efficient reducing power (RP as compared with other plant samples. All the plant extracts exhibited a relative alpha-amylase inhibition apart from Strophanthus hispidus (SH extract with a negative effect. We discovered a mild to weak correlation between alpha-amylase inhibition or antioxidative capacity and the total phenol or flavonoid content. At least in part, the results obtained in this work support the traditional use of certain plant species in the treatment of patients with DM.

  9. Alpha-Amylase Inhibition and Antioxidative Capacity of Some Antidiabetic Plants Used by the Traditional Healers in Southeastern Nigeria

    Science.gov (United States)

    Oyedemi, Blessing O.; Ijeh, Ifeoma I.; Ohanyerem, Princemartins E.; Aiyegoro, Olayinka A.

    2017-01-01

    Oxidative stress plays a significant role in the pathogenesis of metabolic syndrome including diabetes mellitus (DM). The inhibition of alpha-amylase is an important therapeutic target in the regulation of postprandial increase of blood glucose in diabetic patients. The present study investigated the alpha-amylase inhibitory and antioxidant potential of selected herbal drugs used in the treatment of DM by the traditional healers in Isiala Mbano and Ikwuano regions of southeastern Nigeria. Antioxidant activity was evaluated in terms of free radical scavenging, reducing power, and total phenolic (TPC) and flavonoid content (TFC) in consonance with the TLC profiling. The results showed that methanol crude extracts from Anacardium occidentale (AO) and Ceiba pentandra (CP) recorded higher TPC and TFC, potent free radical scavenging, and efficient reducing power (RP) as compared with other plant samples. All the plant extracts exhibited a relative alpha-amylase inhibition apart from Strophanthus hispidus (SH) extract with a negative effect. We discovered a mild to weak correlation between alpha-amylase inhibition or antioxidative capacity and the total phenol or flavonoid content. At least in part, the results obtained in this work support the traditional use of certain plant species in the treatment of patients with DM. PMID:28367491

  10. Alpha amylase assisted synthesis of TiO2 nanoparticles: Structural characterization and application as antibacterial agents

    International Nuclear Information System (INIS)

    Ahmad, Razi; Mohsin, Mohd; Ahmad, Tokeer; Sardar, Meryam

    2015-01-01

    Graphical abstract: - Highlights: • Green synthesis of TiO 2 nanoparticles using an enzyme alpha amylase has been described. • The morphology and shape depends upon the concentration of the alpha amylase enzyme. • The biosynthesized nanoparticles show good bactericidal effect against both gram positive and gram negative bacteria. • The bactericidal effect was further confirmed by Confocal microscopy and TEM. - Abstract: The enzyme alpha amylase was used as the sole reducing and capping agent for the synthesis of TiO 2 nanoparticles. The biosynthesized nanoparticles were characterized by X-ray diffraction (XRD) and transmission electron microscopic (TEM) methods. The XRD data confirms the monophasic crystalline nature of the nanoparticles formed. TEM data shows that the morphology of nanoparticles depends upon the enzyme concentration used at the time of synthesis. The presence of alpha amylase on TiO 2 nanoparticles was confirmed by FTIR. The nanoparticles were investigated for their antibacterial effect on Staphylococcus aureus and Escherichia coli. The minimum inhibitory concentration value of the TiO 2 nanoparticles was found to be 62.50 μg/ml for both the bacterial strains. The inhibition was further confirmed using disc diffusion assay. It is evident from the zone of inhibition that TiO 2 nanoparticles possess potent bactericidal activity. Further, growth curve study shows effect of inhibitory concentration of TiO 2 nanoparticles against S. aureus and E. coli. Confocal microscopy and TEM investigation confirm that nanoparticles were disrupting the bacterial cell wall

  11. Relationship Between Meditation Depth and Waking Salivary Alpha-Amylase Secretion Among Long-Term MBSR Instructors.

    Science.gov (United States)

    Haslam, Alyson; Wirth, Michael D; Robb, Sara Wagner

    2017-08-01

    The purpose of this study was to characterize sympathetic activity by using waking salivary alpha-amylase (sAA) concentrations in a group of long-term meditation instructors and to examine the association between meditation (depth, dose and duration) and the waking alpha-amylase response. Salivary alpha-amylase samples were collected (immediately upon waking and at 15-min, 30-min and 45-min intervals after waking) from mindfulness-based stress reduction instructors to determine both the area under the curve and the awakening slope (difference in alpha-amylase concentrations between waking and 30-min post-waking). It was determined through general linear models that neither years of meditation nor meditation dose were associated with the awakening sAA slope, but higher scores for meditation depth (greater depth) was associated with a more negative (or steeper) awakening slope [Quartile (Q)1: -7 versus Q4: -21 U/mL; p = 0.06], in fully adjusted models. Older age (p = 0.04) and a later time of waking (p meditate more deeply. Copyright © 2016 John Wiley & Sons, Ltd. Copyright © 2016 John Wiley & Sons, Ltd.

  12. Human α-amylase present in lower-genital-tract mucosal fluid processes glycogen to support vaginal colonization by Lactobacillus.

    Science.gov (United States)

    Spear, Gregory T; French, Audrey L; Gilbert, Douglas; Zariffard, M Reza; Mirmonsef, Paria; Sullivan, Thomas H; Spear, William W; Landay, Alan; Micci, Sandra; Lee, Byung-Hoo; Hamaker, Bruce R

    2014-10-01

    Lactobacillus colonization of the lower female genital tract provides protection from the acquisition of sexually transmitted diseases, including human immunodeficiency virus, and from adverse pregnancy outcomes. While glycogen in vaginal epithelium is thought to support Lactobacillus colonization in vivo, many Lactobacillus isolates cannot utilize glycogen in vitro. This study investigated how glycogen could be utilized by vaginal lactobacilli in the genital tract. Several Lactobacillus isolates were confirmed to not grow in glycogen, but did grow in glycogen-breakdown products, including maltose, maltotriose, maltopentaose, maltodextrins, and glycogen treated with salivary α-amylase. A temperature-dependent glycogen-degrading activity was detected in genital fluids that correlated with levels of α-amylase. Treatment of glycogen with genital fluids resulted in production of maltose, maltotriose, and maltotetraose, the major products of α-amylase digestion. These studies show that human α-amylase is present in the female lower genital tract and elucidates how epithelial glycogen can support Lactobacillus colonization in the genital tract. © The Author 2014. Published by Oxford University Press on behalf of the Infectious Diseases Society of America. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.

  13. Antibacterial activity of plasma from crocodile (Crocodylus siamensis against pathogenic bacteria

    Directory of Open Access Journals (Sweden)

    Kommanee Jintana

    2012-07-01

    Full Text Available Abstract Background The Siamese crocodile (Crocodylus siamensis is a critically endangered species of freshwater crocodiles. Crocodilians live with opportunistic bacterial infection but normally suffer no adverse effects. They are not totally immune to microbial infection, but their resistance thereto is remarkably effective. In this study, crude and purified plasma extracted from the Siamese crocodile were examined for antibacterial activity against clinically isolated, human pathogenic bacterial strains and the related reference strains. Methods Crude plasma was prepared from whole blood of the Siamese crocodile by differential sedimentation. The crude plasma was examined for antibacterial activity by the liquid growth inhibition assay. The scanning electron microscopy was performed to confirm the effect of crude crocodile plasma on the cells of Salmonella typhi ATCC 11778. Effect of crude crocodile plasma on cell viability was tested by MTT assay. In addition, the plasma was purified by anion exchange column chromatography with DEAE-Toyopearl 650 M and the purified plasma was tested for antibacterial activity. Results Crude plasma was prepared from whole blood of the Siamese crocodile and exhibited substantial antibacterial activities of more than 40% growth inhibition against the six reference strains of Staphylococcus aureus, Salmonella typhi, Escherichia coli, Vibrio cholerae, Pseudomonas aeruginosa, and Staphylococcus epidermidis, and the four clinical isolates of Staphylococcus epidermidis, Pseudomonas aeruginosa, Salmonella typhi, and Vibrio cholerae. Especially, more than 80% growth inhibition was found in the reference strains of Salmonella typhi, Vibrio cholerae, and Staphylococcus epidermidis and in the clinical isolates of Salmonella typhi and Vibrio cholerae. The effect of the crude plasma on bacterial cells of Salmonella typhi, a certain antibacterial material probably penetrates progressively into the cytoplasmic space

  14. α-Amylase inhibitor-1 gene from Phaseolus vulgaris expressed in Coffea arabica plants inhibits α-amylases from the coffee berry borer pest

    Directory of Open Access Journals (Sweden)

    Oliveira-Neto Osmundo B

    2010-06-01

    Full Text Available Abstract Background Coffee is an important crop and is crucial to the economy of many developing countries, generating around US$70 billion per year. There are 115 species in the Coffea genus, but only two, C. arabica and C. canephora, are commercially cultivated. Coffee plants are attacked by many pathogens and insect-pests, which affect not only the production of coffee but also its grain quality, reducing the commercial value of the product. The main insect-pest, the coffee berry borer (Hypotheneumus hampei, is responsible for worldwide annual losses of around US$500 million. The coffee berry borer exclusively damages the coffee berries, and it is mainly controlled by organochlorine insecticides that are both toxic and carcinogenic. Unfortunately, natural resistance in the genus Coffea to H. hampei has not been documented. To overcome these problems, biotechnological strategies can be used to introduce an α-amylase inhibitor gene (α-AI1, which confers resistance against the coffee berry borer insect-pest, into C. arabica plants. Results We transformed C. arabica with the α-amylase inhibitor-1 gene (α-AI1 from the common bean, Phaseolus vulgaris, under control of the seed-specific phytohemagglutinin promoter (PHA-L. The presence of the α-AI1 gene in six regenerated transgenic T1 coffee plants was identified by PCR and Southern blotting. Immunoblotting and ELISA experiments using antibodies against α-AI1 inhibitor showed a maximum α-AI1 concentration of 0.29% in crude seed extracts. Inhibitory in vitro assays of the α-AI1 protein against H. hampei α-amylases in transgenic seed extracts showed up to 88% inhibition of enzyme activity. Conclusions This is the first report showing the production of transgenic coffee plants with the biotechnological potential to control the coffee berry borer, the most important insect-pest of crop coffee.

  15. Alpha-amylase inhibitor-1 gene from Phaseolus vulgaris expressed in Coffea arabica plants inhibits alpha-amylases from the coffee berry borer pest.

    Science.gov (United States)

    Barbosa, Aulus E A D; Albuquerque, Erika V S; Silva, Maria C M; Souza, Djair S L; Oliveira-Neto, Osmundo B; Valencia, Arnubio; Rocha, Thales L; Grossi-de-Sa, Maria F

    2010-06-17

    Coffee is an important crop and is crucial to the economy of many developing countries, generating around US$70 billion per year. There are 115 species in the Coffea genus, but only two, C. arabica and C. canephora, are commercially cultivated. Coffee plants are attacked by many pathogens and insect-pests, which affect not only the production of coffee but also its grain quality, reducing the commercial value of the product. The main insect-pest, the coffee berry borer (Hypotheneumus hampei), is responsible for worldwide annual losses of around US$500 million. The coffee berry borer exclusively damages the coffee berries, and it is mainly controlled by organochlorine insecticides that are both toxic and carcinogenic. Unfortunately, natural resistance in the genus Coffea to H. hampei has not been documented. To overcome these problems, biotechnological strategies can be used to introduce an alpha-amylase inhibitor gene (alpha-AI1), which confers resistance against the coffee berry borer insect-pest, into C. arabica plants. We transformed C. arabica with the alpha-amylase inhibitor-1 gene (alpha-AI1) from the common bean, Phaseolus vulgaris, under control of the seed-specific phytohemagglutinin promoter (PHA-L). The presence of the alpha-AI1 gene in six regenerated transgenic T1 coffee plants was identified by PCR and Southern blotting. Immunoblotting and ELISA experiments using antibodies against alpha-AI1 inhibitor showed a maximum alpha-AI1 concentration of 0.29% in crude seed extracts. Inhibitory in vitro assays of the alpha-AI1 protein against H. hampei alpha-amylases in transgenic seed extracts showed up to 88% inhibition of enzyme activity. This is the first report showing the production of transgenic coffee plants with the biotechnological potential to control the coffee berry borer, the most important insect-pest of crop coffee.

  16. Preliminary studies of plasma growth hormone releasing activity during medical therapy of acromegaly

    International Nuclear Information System (INIS)

    Hagen, T.C.; Lawrence, A.M.; Kirsteins, L.

    1978-01-01

    The in vitro growth hormone releasing activity of plasma obtained from six acromegalic subjects was measured before and during therapy. In five subjects, plasmas were obtained before and during successful medical therapy with medroxyprogesterone acetate (MPA). The sixth subject was sampled before and after transphenoidal Sr 90 -induced hypopituitarism. All subjects had a decrement in fasting growth hormone levels with respective therapies (29-88%). The in vitro growth hormone released from Rhesus monkey anterior pituitaries was assessed after incubating one lateral half in control plasma (pre-therapy) and the contralateral pituitary half in plasma obtained during or after therapy. Studies with plasmas obtained from the five patients successfully treated with MPA showed a decrease in growth hormone releasing activity during therapy in all (18-57%). Plasma obtained after Sr 90 pituitary ablation in the sixth subject had 35% more growth hormone releasing activity than obtained before therapy. These results suggest that active acromegalics who respond to MPA with significantly lowered growth hormone levels may actually achieve this response because of a decrease in growth hormone releasing factor measured peripherally. The opposite response in one acromegalic subject, following Sr 90 pituitary ablation and hypopituitarism, suggests that growth hormone releasing factor secretion may increase when growth hormone levels are lowered by ablative therapy. (orig.) [de

  17. Mammalian mucosal α-glucosidases coordinate with α-amylase in the initial starch hydrolysis stage to have a role in starch digestion beyond glucogenesis.

    Science.gov (United States)

    Dhital, Sushil; Lin, Amy Hui-Mei; Hamaker, Bruce R; Gidley, Michael J; Muniandy, Anbuhkani

    2013-01-01

    Starch digestion in the human body is typically viewed in a sequential manner beginning with α-amylase and followed by α-glucosidase to produce glucose. This report indicates that the two enzyme types can act synergistically to digest granular starch structure. The aim of this study was to investigate how the mucosal α-glucosidases act with α-amylase to digest granular starch. Two types of enzyme extracts, pancreatic and intestinal extracts, were applied. The pancreatic extract containing predominantly α-amylase, and intestinal extract containing a combination of α-amylase and mucosal α-glucosidase activities, were applied to three granular maize starches with different amylose contents in an in vitro system. Relative glucogenesis, released maltooligosaccharide amounts, and structural changes of degraded residues were examined. Pancreatic extract-treated starches showed a hydrolysis limit over the 12 h incubation period with residues having a higher gelatinization temperature than the native starch. α-Amylase combined with the mucosal α-glucosidases in the intestinal extract showed higher glucogenesis as expected, but also higher maltooligosaccharide amounts indicating an overall greater degree of granular starch breakdown. Starch residues after intestinal extract digestion showed more starch fragmentation, higher gelatinization temperature, higher crystallinity (without any change in polymorph), and an increase of intermediate-sized or small-sized fractions of starch molecules, but did not show preferential hydrolysis of either amylose or amylopectin. Direct digestion of granular starch by mammalian recombinant mucosal α-glucosidases was observed which shows that these enzymes may work either independently or together with α-amylase to digest starch. Thus, mucosal α-glucosidases can have a synergistic effect with α-amylase on granular starch digestion, consistent with a role in overall starch digestion beyond their primary glucogenesis function.

  18. Isolation, identification and in silico analysis of alpha-amylase gene of Aspergillus niger strain CSA35 obtained from cassava undergoing spoilage

    Directory of Open Access Journals (Sweden)

    Oghenetega J. Avwioroko

    2018-07-01

    Full Text Available In this investigation, a gene (CDF_Amyl encoding extracellular α-amylase in Aspergillus niger strain CSA35 associated with cassava spoilage was amplified using specific primers and characterized in silico. The gene had a partial nucleotide sequence of 968 bp and encoded a protein of 222 aa residues with a molecular weight and isoelectric point of 25.13 kDa and 4.17, respectively. Its catalytic site was located in the active site domain. BLASTp analysis showed that the protein primary sequence of the α-amylase gene had 98% and 99% homologies with the α-amylase of A. niger and A. oryzae RIB40, respectively. The gene is more closely related to α-amylase genes from fungi than to bacterial, plant, or animal α-amylase genes. Restriction mapping of the gene showed it can be digested with restriction enzymes like NcoI, PstI, SmaI, and BcLI among others but not with EcoRI and EcoRV. Its protein product had a hydrophobicity score of − 0.43 but no transmembrane helix. The CDF_Amyl protein was subcellularly localized in the secretory pathway, an indication of its release into extracellular space after secretion. Also, the 3D structure of the CDF-Amyl protein was barrel-shaped with domains characteristic of α-amylases. The encoded α-amylase Vmax is 6.90 U/mg protein and Km is 6.70 mg/ml. It was concluded that the unique characteristics of the CDF_Amyl gene and its deduced protein could find applications in biotechnological, food and pharmaceutical industries where cloning and further modification of this gene would be required for product development and improvement.

  19. Mammalian mucosal α-glucosidases coordinate with α-amylase in the initial starch hydrolysis stage to have a role in starch digestion beyond glucogenesis.

    Directory of Open Access Journals (Sweden)

    Sushil Dhital

    Full Text Available Starch digestion in the human body is typically viewed in a sequential manner beginning with α-amylase and followed by α-glucosidase to produce glucose. This report indicates that the two enzyme types can act synergistically to digest granular starch structure. The aim of this study was to investigate how the mucosal α-glucosidases act with α-amylase to digest granular starch. Two types of enzyme extracts, pancreatic and intestinal extracts, were applied. The pancreatic extract containing predominantly α-amylase, and intestinal extract containing a combination of α-amylase and mucosal α-glucosidase activities, were applied to three granular maize starches with different amylose contents in an in vitro system. Relative glucogenesis, released maltooligosaccharide amounts, and structural changes of degraded residues were examined. Pancreatic extract-treated starches showed a hydrolysis limit over the 12 h incubation period with residues having a higher gelatinization temperature than the native starch. α-Amylase combined with the mucosal α-glucosidases in the intestinal extract showed higher glucogenesis as expected, but also higher maltooligosaccharide amounts indicating an overall greater degree of granular starch breakdown. Starch residues after intestinal extract digestion showed more starch fragmentation, higher gelatinization temperature, higher crystallinity (without any change in polymorph, and an increase of intermediate-sized or small-sized fractions of starch molecules, but did not show preferential hydrolysis of either amylose or amylopectin. Direct digestion of granular starch by mammalian recombinant mucosal α-glucosidases was observed which shows that these enzymes may work either independently or together with α-amylase to digest starch. Thus, mucosal α-glucosidases can have a synergistic effect with α-amylase on granular starch digestion, consistent with a role in overall starch digestion beyond their primary

  20. Mammalian Mucosal α-Glucosidases Coordinate with α-Amylase in the Initial Starch Hydrolysis Stage to Have a Role in Starch Digestion beyond Glucogenesis

    Science.gov (United States)

    Dhital, Sushil; Lin, Amy Hui-Mei; Hamaker, Bruce R.; Gidley, Michael J.; Muniandy, Anbuhkani

    2013-01-01

    Starch digestion in the human body is typically viewed in a sequential manner beginning with α-amylase and followed by α-glucosidase to produce glucose. This report indicates that the two enzyme types can act synergistically to digest granular starch structure. The aim of this study was to investigate how the mucosal α-glucosidases act with α-amylase to digest granular starch. Two types of enzyme extracts, pancreatic and intestinal extracts, were applied. The pancreatic extract containing predominantly α-amylase, and intestinal extract containing a combination of α-amylase and mucosal α-glucosidase activities, were applied to three granular maize starches with different amylose contents in an in vitro system. Relative glucogenesis, released maltooligosaccharide amounts, and structural changes of degraded residues were examined. Pancreatic extract-treated starches showed a hydrolysis limit over the 12 h incubation period with residues having a higher gelatinization temperature than the native starch. α-Amylase combined with the mucosal α-glucosidases in the intestinal extract showed higher glucogenesis as expected, but also higher maltooligosaccharide amounts indicating an overall greater degree of granular starch breakdown. Starch residues after intestinal extract digestion showed more starch fragmentation, higher gelatinization temperature, higher crystallinity (without any change in polymorph), and an increase of intermediate-sized or small-sized fractions of starch molecules, but did not show preferential hydrolysis of either amylose or amylopectin. Direct digestion of granular starch by mammalian recombinant mucosal α-glucosidases was observed which shows that these enzymes may work either independently or together with α-amylase to digest starch. Thus, mucosal α-glucosidases can have a synergistic effect with α-amylase on granular starch digestion, consistent with a role in overall starch digestion beyond their primary glucogenesis function. PMID

  1. Active control of internal transport barrier and confinement database in JT-60U reversed shear plasma

    Energy Technology Data Exchange (ETDEWEB)

    Sakamoto, Yoshiteru; Takizuka, Tomonori; Shirai, Hiroshi; Fujita, Takaaki; Kamada, Yutaka; Ide, Shunsuke; Fukuda, Takeshi; Koide, Yoshihiko [Japan Atomic Energy Research Inst., Naka, Ibaraki (Japan). Naka Fusion Research Establishment

    2001-07-01

    Active control of internal transport barrier (ITB) and confinement properties of plasma with ITB have been studied in reversed shear plasmas. Modifications of the radial electric field (E{sub r}) profile by changing the combination of tangential neutral beams can control the ITB strength, where the contribution to E{sub r} from the toroidal rotation plays an important role. The ITB confinement database of reversed shear plasmas has been constructed. Stored energy is strongly correlated with poloidal magnetic field at the ITB foot. (author)

  2. The influence of blood plasma of irradiated animals on activity of Ca2+ - ATPase and Mg2+ - ATPase in plasma membrane of thymocytes

    International Nuclear Information System (INIS)

    Dreval', V.I.

    1994-01-01

    Rats were irradiated at doses 1.5, 4.0, 7.0 and 10 Gy. After 1, 8, 15, 22 and 30 days the effect of blood plasma on activity of Ca 2+ -ATPase and Mg 2+ -ATPase in plasma membrane of thymocytes was investigated. It was found that the raise of irradiation dose leads to increasing of blood plasma effect on membrane-bound enzymes

  3. Plasma properties

    International Nuclear Information System (INIS)

    Weitzner, H.

    1990-06-01

    This paper discusses the following topics: MHD plasma activity: equilibrium, stability and transport; statistical analysis; transport studies; edge physics studies; wave propagation analysis; basic plasma physics and fluid dynamics; space plasma; and numerical methods

  4. Immobilization of Trichoderma harzianum α-amylase on PPyAgNp/Fe3O4-nanocomposite: chemical and physical properties.

    Science.gov (United States)

    Mohamed, Saleh A; Al-Harbi, Majed H; Almulaiky, Yaaser Q; Ibrahim, Ibrahim H; Salah, Hala A; El-Badry, Mohamed O; Abdel-Aty, Azza M; Fahmy, Afaf S; El-Shishtawy, Reda M

    2018-03-26

    In this study, a new support has been developed by immobilization of α-amylase onto modified magnetic Fe 3 O 4 -nanoparticles. The characterization of soluble and immobilized α-amylases with regards to kinetic parameters, pH, thermal stability and reusability was studied. The effect of polypyrrole/silver nanocomposite (PPyAgNp) percentage on weight of Fe 3 O 4 and pH on the immobilization of α-amylase was studied. The highest immobilization efficiency (75%) was detected at 10% PPyAgNp/Fe 3 O 4 -nanocomposite and pH 7.0. Immobilization of α-amylase on PPyAgNp/Fe 3 O 4 -nanocomposite was characterized by FT-IR spectroscopy and scanning electron microscopy. The reusability of the immobilized enzyme activity was 80% of its initial activity after 10 reuses. The immobilized enzyme was more stable towards pH, temperature and metal ions compared with soluble enzyme. The kinetic study appeared higher affinity of immobilized enzyme (K m 2.5 mg starch) compared with soluble enzyme (K m 3.5 mg starch). In conclusion, the immobilization of α-amylase on PPyAgNp/Fe 3 O 4 -nanocomposite could successfully be used in industrial and medical applications.

  5. Plasma Dopamine-Beta-Hydroxylase as an Index of Peripheral Noradrenergic Activity

    Science.gov (United States)

    1981-08-17

    and an acidic buffer are included in the DBH mix along with the substrate. In addition, a monoamine oxidase inhibitor is added to the mix to prevent ...diseases (41 , 155, 97). Patients with hyperthyroidism have significantly lower DBH values than those of controls (190, 192), and patients with hypo... hyperthyroidism , and plasma DBH activity was inversely related to thyroxin levels during therapy for hypothyroidism. Although this and plasma NE

  6. Comprehensive enzymatic analysis of the amylolytic system in the digestive fluid of the sea hare, Aplysia kurodai: Unique properties of two α-amylases and two α-glucosidases

    Directory of Open Access Journals (Sweden)

    Akihiko Tsuji

    2014-01-01

    Full Text Available Sea lettuce (Ulva pertusa is a nuisance species of green algae that is found all over the world. East-Asian species of the marine gastropod, the sea hare Aplysia kurodai, shows a clear feeding preference for sea lettuce. Compared with cellulose, sea lettuce contains a higher amount of starch as a storage polysaccharide. However, the entire amylolytic system in the digestive fluid of A. kurodai has not been studied in detail. We purified α-amylases and α-glucosidases from the digestive fluid of A. kurodai and investigated the synergistic action of these enzymes on sea lettuce. A. kurodai contain two α-amylases (59 and 80 kDa and two α-glucosidases (74 and 86 kDa. The 59-kDa α-amylase, but not the 80-kDa α-amylase, was markedly activated by Ca2+ or Cl−. Both α-amylases degraded starch and maltoheptaose, producing maltotriose, maltose, and glucose. Glucose production from starch was higher with 80-kDa α-amylase than with 59-kDa α-amylase. Kinetic analysis indicated that 74-kDa α-glucosidase prefers short α-1,4-linked oligosaccharide, whereas 86-kDa α-glucosidase prefers large α-1,6 and α-1,4-linked polysaccharides such as glycogen. When sea lettuce was used as a substrate, a 2-fold greater amount of glucose was released by treatment with 59-kDa α-amylase and 74-kDa α-glucosidase than by treatment with 45-kDa cellulase and 210-kDa β-glucosidase of A. kurodai. Unlike mammals, sea hares efficiently digest sea lettuce to glucose by a combination of two α-amylases and two α-glucosidases in the digestive fluids without membrane-bound maltase–glucoamylase and sucrase–isomaltase complexes.

  7. Plasma volume, osmolality, vasopressin, and renin activity during graded exercise in man

    Science.gov (United States)

    Convertino, V. A.; Keil, L. C.; Bernauer, E. M.; Greenleaf, J. E.

    1981-01-01

    The influence of work intensity on plasma volume, osmolality, vasopressin and renin activity and the interrelationships between these responses are investigated. Plasma volume, renin activity and osmotic, sodium and arginine vasopressin concentrations were measured in venous blood samples taken from 15 healthy male subjects before and after six minutes of bicycle ergometer exercise at 100, 175 and 225 W. Plasma volume is found to decrease significantly with increasing work intensity, while increases in Na(+) concentration, osmolality and vasopressin are only observed to be significant when the work intensity exceeds 40% maximal aerobic capacity and plasma resin activity increased linearly at all work levels. In addition, significant correlations are observed between plasma volume and osmolality and sodium changes, and between vasopressin and osmolality and sodium content changes. Data thus support the hypotheses that (1) vasopressin may be the primary controlling endocrine for fluid and electrolyte levels following exercise; (2) an exercise intensity greater than 40% maximal aerobic capacity is required to stimulate vasopressin release through changes in plasma osmolality; and (3) the stimulation of the renin-angiotensin system is a more general stress response.

  8. Exposure-response relations of alpha-amylase sensitisation in British bakeries and flour mills

    OpenAIRE

    Nieuwenhuijsen, M. J.; Heederik, D.; Doekes, G.; Venables, K. M.; Newman, T

    1999-01-01

    OBJECTIVES: To describe the levels of exposure to fungal alpha-amylase in British bakeries and flour mills, and to describe the relation between exposure to alpha-amylase and sensitisation to fungal alpha- amylase. METHODS: 495 personal flour dust samples were taken in seven British bakeries and flour mills and analysed for alpha-amylase with an immunoassay. Workers at the sites were asked to fill out questionnaires on work related symptoms, smoking history, and work history, and they w...

  9. Amylase for Apple Juice Processing: Effects of pH, Heat, and Ca2+ Ions

    Directory of Open Access Journals (Sweden)

    Liliana N. Ceci

    2002-01-01

    Full Text Available The aim of this paper was to evaluate the effects of pH, heat, and Ca2+ ions on the α-amylase activities in a commercial amylolytic enzyme (Tyazyme L300, used for apple juice processing. Kinetics of thermal inactivation was studied in acetate and citrate/phosphate buffers at different temperatures (55–70 °C and enzyme concentrations (0.276 and 0.552 mL/100 mL. Maximum α-amylase activity was observed at pH=3.4 in both buffers. Effects of the addition of calcium chloride during and after thermal treatments were also investigated. α-amylase activities were measured by an iodometric method and thermal inactivation constants and D values (time for reducing 90 % of the enzymatic activity were estimated. The enzyme was more sensible to pH changes and heat when citrate ions were present in the reaction medium. If Ca2+ in the enzyme structure is bound to citrate then the resistance of the enzyme to pH changes and heat is lowered. Kinetics obtained according to Arrhenius equation and two enzymatic fractions (thermo-labile and thermoresistant were observed too. In citrate buffer the following relation was observed for thermo-labile fraction: log (D value = -0.144 t/°C + 12.992. The level of thermal inactivation also depended on the enzyme concentration. Higher thermal inactivation rates were obtained by increasing the enzyme concentration in the case when citrate was present. It was also found that the addition of calcium chloride (1 g/L after thermal treatment in median containing citrate reactivated the enzyme treated at 60 and 65 °C. The possible implications of these findings in apple juice processing were discussed.

  10. Culture medium for amylase production by toxigenic fungi

    Directory of Open Access Journals (Sweden)

    Figueira Edson Luiz Zangrando

    2000-01-01

    Full Text Available Mycelial growth and amylase production by a mycotoxigenic strain of Fusarium moniliforme and Aspergillus flavus were evaluated in a culture medium containing starch, glycerol, wheat bran or corn. With emphasis on corn, different fractions composed by germ, degermed seed, starch, milky stage corn and the respective starch or supernatant fraction were analyzed for F. moniliforme growth . The medium composed of milky stage corn supernatant promoted the best mycelial growth (p<0.05, and it was used to prepare amylase production medium in the next step. The medium composed with 2% ground corn in milky stage corn supernatant (350g of milky stage corn blended with 250mL water and centrifuged promoted the highest amylase production, which was at the 10th day of fermentation, both for F. moniliforme (42.32U/mL and A. flavus (4,745.54U/mL.

  11. Pre-analytical and Analytical Variables Affecting the Measurement of Plasma-Derived Microparticle Tissue Factor Activity

    Science.gov (United States)

    Lee, RD; Barcel, DA; Williams, JC; Wang, JG; Boles, JC; Manly, DA; Key, NS; Mackman, N

    2011-01-01

    Introduction Elevated levels of tissue factor positive (TF+) microparticles (MPs) are observed in plasma from a variety of patients with an increased risk of thrombosis. We and others have described the measurement of TF activity in MPs isolated from plasma. The aim of this study was to investigate the effects of pre-analytical and analytical variables on TF activity of MPs isolated from blood of healthy volunteers treated ex vivo with or without bacterial lipopolysaccharide. Materials and Methods We evaluated the following parameters: use of different centrifugation speeds to isolate the MPs; comparison of TF activity of MPs isolated from platelet poor plasma versus platelet free plasma; effect of freeze/thaw on MP TF activity; and comparison of the MP TF activity assay with the measurement of TF protein by ELISA or flow cytometry. Results MPs prepared from platelet poor plasma by centrifugation at 20,000 × g or 100,000 × g for 15 minutes had similar levels of TF activity. However, significantly less TF activity was found in MPs isolated from platelet free plasma compared with platelet poor plasma. Interestingly, freeze/thawing of the plasma showed donor to donor variation in MP TF activity, with a moderate increase in some individuals. Conclusion TF+ MPs can be quantitatively isolated from platelet poor or platelet free plasma by centrifugation at 20,000 × g for 15 minutes. Measurement of MP TF activity in plasma can be used to detect a prothrombotic state in patients with various diseases. PMID:21737126

  12. Plasma membrane factor XIIIA transglutaminase activity regulates osteoblast matrix secretion and deposition by affecting microtubule dynamics.

    Directory of Open Access Journals (Sweden)

    Hadil F Al-Jallad

    2011-01-01

    Full Text Available Transglutaminase activity, arising potentially from transglutaminase 2 (TG2 and Factor XIIIA (FXIIIA, has been linked to osteoblast differentiation where it is required for type I collagen and fibronectin matrix deposition. In this study we have used an irreversible TG-inhibitor to 'block -and-track' enzyme(s targeted during osteoblast differentiation. We show that the irreversible TG-inhibitor is highly potent in inhibiting osteoblast differentiation and mineralization and reduces secretion of both fibronectin and type I collagen and their release from the cell surface. Tracking of the dansyl probe by Western blotting and immunofluorescence microscopy demonstrated that the inhibitor targets plasma membrane-associated FXIIIA. TG2 appears not to contribute to crosslinking activity on the osteoblast surface. Inhibition of FXIIIA with NC9 resulted in defective secretory vesicle delivery to the plasma membrane which was attributable to a disorganized microtubule network and decreased microtubule association with the plasma membrane. NC9 inhibition of FXIIIA resulted in destabilization of microtubules as assessed by cellular Glu-tubulin levels. Furthermore, NC9 blocked modification of Glu-tubulin into 150 kDa high-molecular weight Glu-tubulin form which was specifically localized to the plasma membrane. FXIIIA enzyme and its crosslinking activity were colocalized with plasma membrane-associated tubulin, and thus, it appears that FXIIIA crosslinking activity is directed towards stabilizing the interaction of microtubules with the plasma membrane. Our work provides the first mechanistic cues as to how transglutaminase activity could affect protein secretion and matrix deposition in osteoblasts and suggests a novel function for plasma membrane FXIIIA in microtubule dynamics.

  13. 21 CFR 184.1012 - α-Amylase enzyme preparation from Bacillus stearothermophilus.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 3 2010-04-01 2009-04-01 true α-Amylase enzyme preparation from Bacillus... GENERALLY RECOGNIZED AS SAFE Listing of Specific Substances Affirmed as GRAS § 184.1012 α-Amylase enzyme preparation from Bacillus stearothermophilus. (a) α-Amylase enzyme preparation is obtained from the culture...

  14. Soluble CD206 plasma levels in rheumatoid arthritis reflect decrease in disease activity

    DEFF Research Database (Denmark)

    Heftdal, Line Dam; Stengaard-Pedersen, Kristian; Ørnbjerg, Lykke Midtbøll

    2017-01-01

    internalization and degradation. The soluble form has been suggested as a biomarker of M2A-macrophage activation. The aim of this study was to investigate sCD206 plasma levels in early RA patients initiating anti-TNFα treatment. Plasma levels of sCD206 were measured by ELISA in samples from 155 early RA patients...... from baseline after 6 months. In the ADA group, however, levels remained lower than baseline throughout the treatment period. In conclusion, initially, plasma sCD206 in early RA patients decreased in accordance with disease activity and initiation of DMARD treatment. Treatment with anti-TNFα preserved......Rheumatoid arthritis (RA) is characterized by chronic joint inflammation and infiltration by activated macrophages. TNFα is a central mediator in this process. The mannose receptor, CD206, is a scavenger receptor expressed by M2A-macrophages and dendritic cells. It is involved in collagen...

  15. Biological activity of alligator, avian, and mammalian insulin in juvenile alligators: plasma glucose and amino acids.

    Science.gov (United States)

    Lance, V A; Elsey, R M; Coulson, R A

    1993-02-01

    The biological activity of alligator, turkey, and bovine insulin on plasma glucose and plasma amino acids was tested in fasted juvenile alligators. Preliminary experiments showed that the stress associated with taking the initial blood sample resulted in a hyperglycemic response lasting more than 24 hr. Despite repeated bleedings no additional hyperglycemic events occurred, and blood glucose declined slowly over the next 7 days. Under these conditions the smallest dose of insulin eliciting a hypoglycemic response was 40 micrograms/kg body wt. A dose of 400 micrograms/kg body wt of either alligator or bovine insulin caused a pronounced hypoglycemia by 12 hr postinjection. Maximum decline in plasma glucose occurred at 24 to 36 hr with a slow return to control levels by 120 hr. There were no significant differences in the hypoglycemic responses to any of the three insulins tested. The decline in plasma amino acids was much more rapid than the decline in plasma glucose in response to insulin. Even at the 40 micrograms/kg body wt dose a significant difference from saline-injected control was seen at 2 hr postinjection. Maximum decline in plasma amino acids occurred at 8 to 12 hr with a return to baseline by 36 hr. These results show that the relatively conservative changes in the sequence of alligator insulin (three amino acid substitutions in the B-chain compared with that of chicken) have little effect on biological activity and that alligator insulin receptors do not appear to discriminate among the three insulins.

  16. Production of surfactant and detergent-stable, halophilic, and alkalitolerant alpha-amylase by a moderately halophilic Bacillus sp. Strain TSCVKK.

    Science.gov (United States)

    Kiran, Kondepudi Kanthi; Chandra, T S

    2008-01-01

    A moderately halophilic alkalitolerant Bacillus sp. Strain TSCVKK, with an ability to produce extracellular halophilic, alkalitolerant, surfactant, and detergent-stable alpha-amylase was isolated from soil samples obtained from a salt-manufacturing industry in Chennai. The culture conditions for higher amylase production were optimized with respect to NaCl, substrate, pH, and temperature. Maximum amylase production of 592 mU/ml was achieved in the medium at 48 h with 10% NaCl, 1% dextrin, 0.4% yeast extract, 0.2% tryptone, and 0.2% CaCl(2) at pH 8.0 at 30 degrees C. The enzyme activity in the culture supernatant was highest with 10% NaCl at pH 7.5 and 55 degrees C. The amylase that was partially purified by acetone precipitation was highly stable in various surfactants and detergents. Glucose, maltose, and maltooligosaccharides were the main end products of starch hydrolysis indicating that it is an alpha-amylase.

  17. Plasma cholesteryl ester transfer protein mass and phospholipid transfer protein activity are associated with leptin in type 2 diabetes mellitus

    NARCIS (Netherlands)

    Dullaart, R. P. F.; de Vries, R.; Dallinga-Thie, G. M.; van Tol, A.; Sluiter, W. J.

    Adipose tissue contributes to plasma levels of lipid transfer proteins and is also the major source of plasma adipokines. We hypothesized that plasma cholesteryl ester transfer protein (CETP) mass, phospholipid transfer protein (PLTP) activity and cholesteryl ester transfer (CET, a measure of CETP

  18. 2003 activity report of the development and research line in controlled thermonuclear fusion of the Plasma Associated Laboratory

    International Nuclear Information System (INIS)

    Ludwig, Gerson Otto

    2004-01-01

    This document represents the 2003 activity report of the development and research line in controlled thermonuclear fusion of the Plasma Associated Laboratory - Brazil, approaching the areas of toroidal systems for magnetic confinement, plasma heating, current generation and high temperature plasma diagnostic

  19. Discovering an Accessible Enzyme: Salivary [alpha]-Amylase--"Prima Digestio Fit in Ore"--A Didactic Approach for High School Students

    Science.gov (United States)

    Marini, Isabella

    2005-01-01

    Human salivary [alpha]-amylase is used in this experimental approach to introduce biology high school students to the concept of enzyme activity in a dynamic way. Through a series of five easy, rapid, and inexpensive laboratory experiments students learn what the activity of an enzyme consists of: first in a qualitative then in a semi-quantitative…

  20. Receptor kinase-mediated control of primary active proton pumping at the plasma membrane

    DEFF Research Database (Denmark)

    Fuglsang, Anja Thoe; Kristensen, Astrid; Cuin, Tracey A.

    2014-01-01

    Acidification of the cell wall space outside the plasma membrane is required for plant growth and is the result of proton extrusion by the plasma membrane-localized H+-ATPases. Here we show that the major plasma membrane proton pumps in Arabidopsis, AHA1 and AHA2, interact directly in vitro...... and in planta with PSY1R, a receptor kinase of the plasma membrane that serves as a receptor for the peptide growth hormone PSY1. The intracellular protein kinase domain of PSY1R phosphorylates AHA2/AHA1 at Thr-881, situated in the autoinhibitory region I of the C-terminal domain. When expressed in a yeast...... heterologous expression system, the introduction of a negative charge at this position caused pump activation. Application of PSY1 to plant seedlings induced rapid in planta phosphorylation at Thr-881, concomitant with an instantaneous increase in proton efflux from roots. The direct interaction between AHA2...

  1. Immobilization of α-amylase onto poly(glycidyl methacrylate) grafted electrospun fibers by ATRP

    International Nuclear Information System (INIS)

    Oktay, Burcu; Demir, Serap; Kayaman-Apohan, Nilhan

    2015-01-01

    In this study, novel α-amylase immobilized poly(vinyl alcohol) (PVA) nanofibers were prepared. The PVA nanofiber surfaces were functionalized with 2-bromoisobutyryl bromide (BiBBr) and followed by surface initiated atom transfer radical polymerization (SI-ATRP) of glycidyl methacrylate (GMA). The morphology of the poly(glycidyl methacrylate) (PGMA) grafted PVA nanofibers was characterized by scanning electron microscopy (SEM). Also PGMA brushes were confirmed by X-ray photo electron microscopy (XPS). α-Amylase was immobilized in a one step process onto the PGMA grafted PVA nanofiber. The characteristic properties of the immobilized and free enzymes were examined. The thermal stability of the enzyme was improved and showed maximum activity at 37 °C by immobilization. pH values of the maximum activity of the free and immobilized enzymes were also found at 6.0 and 6.5, respectively. Free enzyme lost its activity completely within 15 days. The immobilized enzyme lost only 23.8% of its activity within 30 days. - Highlights: • Electrospun photocrosslinkable PVA nanofiber was prepared. • PGMA brushes were conducted on PVA nanofiber via SI-ATRP. • The immobilized enzyme showed maximum activity at pH 6.0 and at 37 °C. • Functionalized nanofibers may be used as promising supports for enzyme immobilization

  2. Associations of geomagnetic activity with plasma sheet thinning and expansion: A statistical study

    International Nuclear Information System (INIS)

    Hones, E.W. Jr.; Pytte, T.; West, H.I. Jr.

    1984-01-01

    Associations of geomagnetic activity in the auroral zone with thinnings and expansions of the magnetotail plasma sheet are examined statistically in this paper. We first identified many plasma sheet thinnings and expansions in plasma and particle data from VELA satellites and from OGO 5 without reference to the ground magnetic data. These events were grouped according to the location of the detecting satellite in the magnetotail. For each such group the times of thinning or expansion were then used as fiducial times in a superposed-epoch analysis of the geomagnetic AL index values that were recorded in 8-hour intervals centered on the event times. The results show that many plasma sheet thinnings and expansions are related to discrete negative bay structures that are the classical signature of substorms. Furthermore, they support earlier findings that plasma sheet thinning and expansion at the VELA orbit (rroughly-equal18 R/sub E/) tend to be associated with the onset of the auroral zone negative bay and the beginning of its subsidence, respectively. Earthward of rroughly-equal13-15 R/sub E/, plasma sheet expansion occurs near the time of the onset of the negative bay, again in agreement with earlier findings. A large fraction of plasma sheet expansions to half thicknesses of > or approx. =6 R/sub E/ at the VELA orbit are associated not with a baylike geomagnetic disturbance but with subsidence of a prolonged interval of disturbance. The study also shows that many plasma sheet expansions are related simply to generally enhanced geomagnetic activity showing no baylike or other distinctive features

  3. PLASMA INSULIN AND IGF-1 AND HEPATIC ACTIVITY IN SAANEN GOAT KIDS, AROUND WEANING

    Directory of Open Access Journals (Sweden)

    Damiano Magistrelli

    2009-02-01

    Full Text Available Weaning is a crucial event in the life of young ruminants. At weaning ruminal and digestive activity are still incomplete, so weaning may coincide with a period of growth stasis. Since insulin and insulin-like growth factor 1 (IGF-1 can play a fundamental role in post-natal development, the aim of the present study was to evaluate plasma variations of insulin and IGF-1 levels and their relationships with the hepatic activity, around weaning.For this purpose, eleven 3-days-old Saanen goat kids were randomly divided into MILK (6 animals and WMIX (5 animals groups. All kids were fed goat milk to age 29 days. After that, MILK kids continued to receive milk, while WMIX ones underwent weaning, based on the progressive replacement of milk with solid feed. WMIX kids were completely weaned on day 48. Blood samples were weekly analyzed for metabolic traits, insulin and IGF-1 levels, alanine aminotransferase (ALT and aspartate aminotransferase (AST activities. On day 50, all animals were slaughtered, liver weight was recorded and liver samples were analyzed for DNA, RNA, phospholipids, glicogen and soluble protein content, ALT and AST activity.On day 50, plasma insulin and IGF-1 were lower in WMIX group, as possible consequence of the lower plasma glucose and amino acids levels. Liver weight was not different between groups, but liver weight expressed as percentage of body weight was lower in WMIX kids and highly correlated to plasma IGF-1. Liver glycogen was also lower in WMIX kids, as possible consequence of the lower plasma glucose.Hepatic ALT and AST activities were not different between groups and both were strongly correlated to plasma insulin. Moreover, insulin was positively correlated to the proteosynthetic capability per cell (RNA/DNA of the liver.Our results indicate that the adopted livestock practice permitted the normal development of the animal used, avoiding growth stasis. Anyway, weaning altered plasma insulin and IGF-1, without affecting

  4. Biochemical studies of amylase, lipase and protease in Callosobruchus maculatus (Coleoptera: Chrysomelidae) populations fed with Vigna unguiculata grain cultivated with diazotrophic bacteria strains.

    Science.gov (United States)

    Silva, L B; Torres, É B; Nóbrega, R A S; Lopes, G N; Vogado, R F; Pavan, B E; Fernandes-Junior, P I

    2017-12-01

    The objective of this study was to evaluate the enzymatic activity of homogenates of insects fed on grain of cowpea, Vigna unguiculata (L.), cultivars grown with different nitrogen sources. For the experiment we used aliquots of the homogenate of 100 unsexed adult insects, emerged from 10 g of grain obtained from four cowpea cultivars: 'BRS Acauã', 'BRS Carijó', 'BRS Pujante', and 'BRS Tapaihum' grown under different regimes of nitrogen sources: mineral fertilizer, inoculation with strains of diazotrophs (BR 3267, BR 3262, BR 3299; INPA 03-11B, 03-84 UFLA, as well as the control (with soil nitrogen). The parameters evaluated were enzymatic activities of insect protease, amylase and lipase and the starch content of the grains. There were differences in the enzymatic activity of amylase, lipase and protease of insect homogenate according to the food source. A lower activity of the enzyme amylase from C. maculatus homogenate was observed when insects were fed grain of the cultivar BRS Carijó. A lower activity of lipase enzyme from C. maculatus homogenate was observed when the insects fed on grain from the interaction of the cultivar Tapaihum inoculated with BR 3262 diazotrophs. The lowest proteolytic activity was observed in homogenate of insects fed on interaction of 'BRS Carijó' inoculated with BR 3262 diazotrophs. Starch content correlated positively with the amylase activity of C. maculatus homogenate. The cultivar BRS Carijó had a different behavior from the other cultivars, according to the cluster analysis.

  5. Specific, sensitive, precise, and rapid functional chromogenic assay of activated first complement component (C1) in plasma

    DEFF Research Database (Denmark)

    Munkvad, S; Jespersen, J; Sidelmann, Johannes Jakobsen

    1990-01-01

    We present a new functional assay for the first complement component (C1) in plasma, based on its activation by inhibition of the C1-esterase inhibitor (C1-inh) when monospecific antiserum to C1-inh is added to the plasma. After maximal activation, we can determine the concentration of activated ...

  6. Depression and Anxiety are Common in Acute HIV Infection and Associate with Plasma Immune Activation.

    Science.gov (United States)

    Hellmuth, Joanna; Colby, Donn; Valcour, Victor; Suttichom, Duanghathai; Spudich, Serena; Ananworanich, Jintanat; Prueksakaew, Peeriya; Sailasuta, Napapon; Allen, Isabel; Jagodzinski, Linda L; Slike, Bonnie; Ochi, Derek; Paul, Robert

    2017-11-01

    This observational study of 123 Thai participants sought to determine the rate and severity of affective symptoms during acute HIV infection (AHI) and possible associations to disease mechanisms. At diagnosis, just prior to starting combination antiretroviral therapy (cART), AHI participants completed assessments of depression and anxiety symptoms that were repeated at 4, 12, and 24 weeks. Blood markers of HIV infection and immune activation were measured at study entry, with optional cerebrospinal fluid measures. A high frequency of participants reported symptoms that exceeded published thresholds supportive of depression (55.0%) and anxiety (65.8%) at diagnosis, with significant reductions after starting cART. Meeting a threshold for clinically relevant depressive symptoms at study entry was associated with higher baseline plasma HIV RNA (5.98 vs. 5.50, t = 2.46, p = 0.015), lower CD4 counts (328 vs. 436 cells/mm 3 , t = 3.46, p = 0.001), and higher plasma neopterin, a marker of macrophage activation (2694 vs. 1730 pg/mL, Mann-Whitney U = 152.5, p = 0.011). Controlling for plasma HIV RNA and CD4 count, higher baseline plasma neopterin correlated with worse initial depression and anxiety scores. Depression and anxiety symptoms are frequent in acute HIV infection, associate with plasma immune activation, and can improve concurrent with cART.

  7. EFFECTS OF CIGARETTE SMOKING ON ERYTHROCYTE ANTIOXIDATIVE ENZYME ACTIVITIES AND PLASMA CONCENTRATIONS OF THEIR COFACTORS

    Directory of Open Access Journals (Sweden)

    M. Zahraie

    2005-07-01

    Full Text Available Tobacco smoke contains numerous compounds, many ‎of which are oxidants and capable of producing free radical and enhancing ‎the oxidative stress. The aim of this study was to investigate the effect of cigarette smoking on the erythrocyte antioxidative enzyme activities and the plasma ‎concentration of their cofactors. ‎Sixty eight healthy men were enrolled, 32 of whom had never smoked and 36 had smoked at least 10 cigarettes per day for ‎at least one year. Hemolysate superoxide dismutase (Cu-Zn SOD, glutathione peroxidase (GSH-Px and ‎catalase (CAT activities were measured using spectrophotometer. Plasma copper, zinc and selenium concentrations were determined ‎using atomic absorption spectrophotometer. Plasma iron concentration was determined by colorimetric ‎method. We found that erythrocyte Cu-Zn SOD activity was significantly higher in tobacco smokers ‎compared with non-smokers (1294 ± 206.7 U/gHb in smokers vs. 1121.6 ± 237.8 U/gHb in non-‎smokers, P < 0.01. While plasma selenium concentration was significantly lower in tobacco ‎smokers (62.7±14.8 μg/L in smokers vs. 92.1 ± 17.5 μg/L in non-smokers, P < 0.01, there were no significant ‎differences in erythrocyte GSH-Px and CAT activities and plasma copper, zinc and iron concentrations between the two groups. ‎It seems that cigarette smoking can alter antioxidative enzymes activity and plasma concentration of some trace elements.

  8. Tissue Factor Coagulant Activity is Regulated by the Plasma Membrane Microenvironment.

    Science.gov (United States)

    Yu, Yuanjie; Böing, Anita N; Hau, Chi M; Hajji, Najat; Ruf, Wolfram; Sturk, Auguste; Nieuwland, Rienk

    2018-06-01

     Tissue factor (TF) can be present in a non-coagulant and coagulant form. Whether the coagulant activity is affected by the plasma membrane microenvironment is unexplored.  This article studies the presence and coagulant activity of human TF in plasma membrane micro-domains.  Plasma membranes were isolated from human MIA PaCa2 cells, MDA-MB-231 cells and human vascular smooth muscle cells by Percoll gradient ultracentrifugation after cell disruption. Plasma membranes were fractionated by OptiPrep gradient ultracentrifugation, and the presence of TF, flotillin, caveolin, clathrin, protein disulphide isomerase (PDI), TF pathway inhibitor (TFPI) and phosphatidylserine (PS) were determined.  Plasma membranes contain two detergent-resistant membrane (DRM) compartments differing in density and biochemical composition. High-density DRMs (DRM-H) have a density ( ρ ) of 1.15 to 1.20 g/mL and contain clathrin, whereas low-density DRMs (DRM-L) have a density between 1.09 and 1.13 g/mL and do not contain clathrin. Both DRMs contain TF, flotillin and caveolin. PDI is detectable in DRM-H, TFPI is not detectable in either DMR-H or DRM-L and PS is detectable in DRM-L. The DRM-H-associated TF (> 95% of the TF antigen) lacks detectable coagulant activity, whereas the DRM-L-associated TF triggers coagulation. This coagulant activity is inhibited by lactadherin and thus PS-dependent, but seemed insensitive to 16F16, an inhibitor of PDI.  Non-coagulant and coagulant TF are present within different types of DRMs in the plasma membrane, and the composition of these DRMs may affect the TF coagulant activity. Schattauer GmbH Stuttgart.

  9. Barley alpha-amylase/subtilisin inhibitor: structure, biophysics and protein engineering

    DEFF Research Database (Denmark)

    Nielsen, P.K.; Bønsager, Birgit Christine; Fukuda, Kenji

    2004-01-01

    Bifunctional alpha-amylase/subtilisin inhibitors have been implicated in plant defence and regulation of endogenous alpha-amylase action. The barley alpha-amylase/subtilisin inhibitor (BASI) inhibits the barley alpha-amylase 2 (AMY2) and subtilisin-type serine proteases. BASI belongs to the Kunitz...... Ca2+-modulated kinetics of the AMY2/BASl interaction and found that the complex formation involves minimal structural changes. The modulation of the interaction by calcium ions makes it unique among the currently known binding mechanisms of proteinaceous alpha-amylase inhibitors....

  10. Thermophilic and unusually acidophilic amylase produced by a thermophilic acidophilic bacillus sp

    Energy Technology Data Exchange (ETDEWEB)

    Uchino, F

    1982-01-01

    Bacillus sp. 11-1S, a thermophilic acidophilic bacterial strain, produced an extracellular amylase with unusual characteristics. The enzyme was purified 40-fold by SE-Sephadex column chromatography. The pH optimum for activity was 2.0, and substantial activity was noted in the pH range of 1.5-3.5. The optimal temperature was 70 degrees C, but the activity decreased markedly in lower reaction temperatures. Arrhenius plots of the reaction showed two straight lines intersecting at about 50 degrees C. The activity or stability of the enzyme was not likely to depend on Ca2+. The molecular weight of the enzyme was 54,000 calculated from the electrophoretic mobility. The enzyme behaved like an alpha-amylase (1,4-alpha-D- glucan glucanohydrolase, E.C. 3.2.1.1). About 34% of glucosidic linkages of soluble starch was hydrolyzed at 65 degrees C and pH 2.0, in 24 hours, and the major products were maltotriose and maltose. (Refs. 14).

  11. Regulation of hepatic lipase activity by sphingomyelin in plasma lipoproteins.

    Science.gov (United States)

    Yang, Peng; Subbaiah, Papasani V

    2015-10-01

    Hepatic lipase (HL) is an important enzyme in the clearance of triacylglycerol (TAG) from the circulation, and has been proposed to have pro-atherogenic as well as anti-atherogenic properties. It hydrolyzes both phospholipids and TAG of lipoproteins, and its activity is negatively correlated with HDL levels. Although it is known that HL acts preferentially on HDL lipids, the basis for this specificity is not known, since it does not require any specific apoprotein for activity. In this study, we tested the hypothesis that sphingomyelin (SM), whose concentration is much higher in VLDL and LDL compared to HDL, is an inhibitor of HL, and that this could explain the lipoprotein specificity of the enzyme. The results presented show that the depletion of SM from normal lipoproteins activated the HL roughly in proportion to their SM content. SM depletion stimulated the hydrolysis of both phosphatidylcholine (PC) and TAG, although the PC hydrolysis was stimulated more. In the native lipoproteins, HL showed specificity for PC species containing polyunsaturated fatty acids at sn-2 position, and produced more unsaturated lyso PC species. The enzyme also showed preferential hydrolysis of certain TAG species over others. SM depletion affected the specificity of the enzyme towards PC and TAG species modestly. These results show that SM is a physiological inhibitor of HL activity in lipoproteins and that the specificity of the enzyme towards HDL is at least partly due to its low SM content. Published by Elsevier B.V.

  12. The use of a-methyl-D-glucoside, a synthetic analogue of maltose, as inducer of amylase by Aspergillus sp in solid-state and submerged fermentations

    Directory of Open Access Journals (Sweden)

    Moreira Fabiana G.

    2001-01-01

    Full Text Available The use of a methyl-D-glucoside (alphaMG, a synthetic analogue of maltose, as carbon source and inducer of amylase synthesis to several species of Aspergillus was studied in submerged and solid-state fermentations. Among a group of ten species, A. tamarii, A. fumigatus and A. flavus were able to produce biomass and high specific amylolytic activity in submerged cultures containing alphaMG as the only carbon source. In solid state fermentation, the enrichment of basal wheat bran or corn cob medium with alphaMG increased up to 3 times the production of amylases. In both submerged and solid state fermentations, alphaMG was more effective inducer of amylases than maltose and starch.

  13. The use of alpha-methyl-D-glucoside, a synthetic analogue of maltose, as inducer of amylase by Aspergillus sp in solid-state and submerged fermentations

    Directory of Open Access Journals (Sweden)

    Fabiana G. Moreira

    2001-03-01

    Full Text Available The use of a methyl-D-glucoside (alphaMG, a synthetic analogue of maltose, as carbon source and inducer of amylase synthesis to several species of Aspergillus was studied in submerged and solid-state fermentations. Among a group of ten species, A. tamarii, A. fumigatus and A. flavus were able to produce biomass and high specific amylolytic activity in submerged cultures containing alphaMG as the only carbon source. In solid state fermentation, the enrichment of basal wheat bran or corn cob medium with alphaMG increased up to 3 times the production of amylases. In both submerged and solid state fermentations, alphaMG was more effective inducer of amylases than maltose and starch.

  14. Effect on hemo-dialysis on concentration of lactate dehyrogenase, creatine kinase and a-amylase in renal failure

    International Nuclear Information System (INIS)

    Modawe, G. O. H.; Idris, O. F.

    2009-01-01

    This study was conducted to compare the concentration of plasma enzymes in chronic renal failure pre dialysis (group A) and post dialysis (group B), and evaluate the concentration of these enzymes between pre and post dialysis. The study was performed in 25 samples of Sudanese patients (chronic renal failure) and compared with 15 samples as the control groups. plasma samples were analyzed using spectrophotometric methods, plasma concentration of these enzymes showed increase in chronic renal failure pre and post dialysis. The mean values of lactate dehydrogenase pre dialysis was 259IU/L, post dialysis was 276IU/L, the mean of creatine kinase pre dialysis was 252IU/L, and post dialysis was 241 IU/L but the mean of amylase pre and post dialysis was the same 144 IU/L. This study showed that there was no difference of concentration of amylase enzyme, but the different in concentration of CK and LDH between pre and post dialysis during chronic renal failure depend on normal range of this enzyme in control groups.(Author)

  15. Oxidation of limonene using activated carbon modified in dielectric barrier discharge plasma

    Science.gov (United States)

    Glonek, Karolina; Wróblewska, Agnieszka; Makuch, Edyta; Ulejczyk, Bogdan; Krawczyk, Krzysztof; Wróbel, Rafał. J.; Koren, Zvi C.; Michalkiewicz, Beata

    2017-10-01

    The waste from industrial fruits processing is utilized for the extraction of limonene, a renewable terpene biomass compound obtained from orange peels. This was followed by limonene oxidation, which produces highly useful oxygenated derivatives (carveol, and perillyl alcohol, 1,2-epoxylimonene and its diol). New catalysts were obtained by treating relatively inexpensive commercially available EuroPh and FPV activated carbons with plasma. These catalysts were characterized by the following instrumental methods XRD, sorption of N2 and CO2, SEM, EDS, TEM, XPS, and Raman spectroscopy. The activities of the plasma-treated catalysts were measured in the oxidation of limonene by means of either hydrogen peroxide or t-butyl hydroperoxide as the oxidizing agents. During the oxidation with hydrogen peroxide the new plasma-treated catalysts were more active than their untreated counterparts. This effect was noticeable in the considerable increase in the conversion of limonene. The mechanism explaining this property is proposed, and it takes into account the role of the appropriate functional groups on the surface of the catalysts. This work has shown for the first time that the commercial EuroPh and FPV activated carbons, after having been treated by plasma, are active catalysts for the selective limonene oxidation for the production of value-added industrial products.

  16. Radioreceptor assay for evaluation of the plasma glucocorticoid activity of natural and synthetic steroids in man

    International Nuclear Information System (INIS)

    Ballard, P.L.; Carter, J.P.; Graham, B.S.; Baxter, J.D.

    1975-01-01

    An assay for plasma glucocorticoid activity has been developed using specific glucocorticoid receptors. Unlike other assays for cortisol and certain synthetic corticosteroids, this radioreceptor assay measures the glucocorticoid activity of all natural and synthetic steroids. Steroids extracted from as little as 0.05 ml of plasma are incubated with 3 H-dexamethasone and cytosol receptors from cultured rat hepatoma cells. From 0.5 to 50 ng of cortisol are accurately detected. Glucocorticoid activities of adult plasmas determined by the assay correlate closely with corticoid levels obtained in the CBG-isotope and fluorometric assays. Other steroids are measured in proportion to both concentration and potency as glucocorticoids. Relative activities include: cortisol 100, dexamethasone 940, prednisolone 230, prednisone 3, estradiol 1 and androstenedione 1. A similar ranking of steroids was found using receptors from a human source (fetal lung). The assay has been useful in detecting glucocorticoid activity in unidentified medications and in measuring plasma glucocorticoid levels after administration of synthetic corticosteroids. (auth)

  17. Recognition of acidic phospholipase A2 activity in plasma membranes of resident peritoneal macrophages

    International Nuclear Information System (INIS)

    Shibata, Y.; Abiko, Y.; Ohno, H.; Araki, T.; Takiguchi, H.

    1988-01-01

    Phospholipase (PLase) activities in the plasma membrane of guinea pig peritoneal macrophages were studied, as these enzymes having such activity may be candidates for the release of arachidonic acid (AA) from phosphatidylcholine (PC). An AA release system operating at acidic pH was identified in the macrophage plasma membrane and characterized. This membrane-bound acidic PLase A 2 had an optimum pH at 4.5, and enzyme activation was observed in Ca ++ -free medium; but the maximum activity was found at 0.5 mM Ca ++ concentration. The Km value for PC of acidic PLase A 2 was 4.2 μM, and a Michaelis-Menten relationship was evident. Calcium might act as a cofactor at some intermediate step during the activation of acidic PLase A 2 in light of the uncompetitive manner of Ca ++ action. Furthermore, the release of [ 3 H]-AA from preradiolabelled macrophage plasma membranes occurred with the addition of Ca ++ at pH 4.5. These data suggest that the acid PLase A 2 is a component of the plasma membrane and is not due to lysosomal contamination since membrane-bound acidic PLase A 2 properties are opposite to those found for lysosomal PLase A 2

  18. Purification and characterization of an alpha-amylase of Pichia burtonii isolated from the traditional starter "murcha" in Nepal.

    Science.gov (United States)

    Takeuchi, Akiko; Shimizu-Ibuka, Akiko; Nishiyama, Yoshitaka; Mura, Kiyoshi; Okada, Sanae; Tokue, Chiyoko; Arai, Soichi

    2006-12-01

    Among more than 20 yeast strains isolated from the traditional starter "murcha" in Nepal, we characterized a yeast that might be involved in saccharification. This strain, identified as Pichia burtonii, produced an extracellular amylolytic enzyme when cultured in the presence of starch in the medium. Since no amylase secreted by P. burtonii has yet been reported, we purified the enzyme and determined its N-terminal amino acid sequence. Together with the results of a hydrolyzing activity assay toward various substrates, it was found to be an alpha-amylase. The purified enzyme, named Pichia burtonii alpha-amylase (PBA), was a glycoprotein with an apparent molecular mass of 51 kDa. Enzyme activity was optimal at pH 5.0 at 40 degrees C. The enzyme retained 80% of its original activity after incubation under the optimal pH condition at 50 degrees C for 30 min. The activity was inhibited by metal ions such as Cd(2+), Cu(2+), Hg(2+), Al(3+), and Zn(2+).

  19. Study on the immobilization of alpha-amylase by radiation-induced polymerization at low-temperature, (2)

    International Nuclear Information System (INIS)

    Yoshida, Masaru; Kumakura, Minoru; Kaetsu, Isao

    1975-07-01

    The immobilization α-amylase in low concentration (50-250μg) by radiation induced polymerization at low temperature, with HEMA has been studied. The immobilization was performed in the temperature range of -196 0 C to +40 0 C. Activity of the immobilized enzyme decreases at temperatures above 0 0 C. The optimum temperatures for immobilization of α-amylase are -78 0 C - -24 0 C, where only the polymerization by irradiation is effective. HEMA is a suitable monomer as the immobilization carrier, because of its high polymerization rate of 100% in the temperature range. The suitable concentration of HEMA is less than 30%, and above this concentration the activity of enzyme decreases considerably. The optimum irradiation dose for immobilization is 1 x 10 6 R, and the activity of enzyme decreases at 5 x 10 6 R. The polymerization composition is porous gel structure, so the enzymatic reaction can be carried out merely by introducing a substrate to the composition. The activity attained in the immobilized enzyme is 75-80% that of the native α-amylase. The immobilized enzyme is more heat-resistant than the native one. (auth.)

  20. Immobilization of α-Amylase from Anoxybacillus sp. SK3-4 on ReliZyme and Immobead Supports

    Directory of Open Access Journals (Sweden)

    Ummirul Mukminin Kahar

    2016-09-01

    Full Text Available α-Amylase from Anoxybacillus sp. SK3-4 (ASKA is a thermostable enzyme that produces a high level of maltose from starches. A truncated ASKA (TASKA variant with improved expression and purification efficiency was characterized in an earlier study. In this work, TASKA was purified and immobilized through covalent attachment on three epoxide (ReliZyme EP403/M, Immobead IB-150P, and Immobead IB-150A and an amino-epoxide (ReliZyme HFA403/M activated supports. Several parameters affecting immobilization were analyzed, including the pH, temperature, and quantity (mg of enzyme added per gram of support. The influence of the carrier surface properties, pore sizes, and lengths of spacer arms (functional groups on biocatalyst performances were studied. Free and immobilized TASKAs were stable at pH 6.0–9.0 and active at pH 8.0. The enzyme showed optimal activity and considerable stability at 60 °C. Immobilized TASKA retained 50% of its initial activity after 5–12 cycles of reuse. Upon degradation of starches and amylose, only immobilized TASKA on ReliZyme HFA403/M has comparable hydrolytic ability with the free enzyme. To the best of our knowledge, this is the first report of an immobilization study of an α-amylase from Anoxybacillus spp. and the first report of α-amylase immobilization using ReliZyme and Immobeads as supports.