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Sample records for bacillus halmapalus alpha-amylase

  1. Factors affecting the solubility of Bacillus halmapalus alpha-amylase

    DEFF Research Database (Denmark)

    Faber, Cornelius; Hobley, Timothy John; Mollerup, Jørgen

    2008-01-01

    A detailed study of the solubility of recombinant Bacillus halmapalus alpha-amylase has been conducted. A semi-purified preparation from a bulk crystallisation was chos en that contained six isoforms with pI-values of between 5.5 and 6.1. The solubility was strongly affected by pH and could...

  2. Structure of Bacillus halmapalus alpha-amylase crystallized with and without the substrate analogue acarbose and maltose

    DEFF Research Database (Denmark)

    Lyhne-Iversen, Louise; Hobley, Timothy John; Kaasgaard, Svend G.

    2006-01-01

    Recombinant Bacillus halmapalus alpha-amylase (BHA) was studied in two different crystal forms. The first crystal form was obtained by crystallisation of BHA at room temperature in the presence of acarbose and maltose - data was collected at cryogenic temperatures to a resolution of 1.9 Å...

  3. A Proposed Mechanism for the Thermal Denaturation of a Recombinant Bacillus Halmapalus Alpha-amylase - the Effect of Calcium Ions

    Science.gov (United States)

    Nielsen, Anders D.; Pusey, Marc L.; Fuglsang, Claus C.; Westh, Peter

    2003-01-01

    The thermal stability of a recombinant alpha-amylase from Bacillus halmapalus alpha-amylase (BHA) has been investigated using circular dichroism spectroscopy (CD) and differential scanning calorimetry (DSC). This alpha-amylase is homologous to other Bacillus alpha-amylases where previous crystallographic studies have identified the existence of 3 calcium binding sites in the structure. Denaturation of BHA is irreversible with a Tm of approximately 89 C, and DSC thermograms can be described using a one-step irreversible model. A 5 C increase in T(sub m) in the presence of 10 fold excess CaCl2 was observed. However, a concomitant increase in the tendency to aggregate was also observed. The presence of 30-40 fold excess calcium chelator (EDTA or EGTA) results in a large destabilization of BHA corresponding to about 40 C lower T(sub m), as determined by both CD and DSC. Ten fold excess EGTA reveals complex DSC thermograms corresponding to both reversible and irreversible transitions, which possibly originate from different populations of BHA:calcium complexes. The observations in the present study have, in combination with structural information of homologous alpha-amylases, provided the basis for the proposal of a simple denaturation mechanism of BHA. The proposed mechanism describes the irreversible thermal denaturation of different BHA:calcium complexes and the calcium binding equilibrium involved. Furthermore, the model accounts for a temperature induced reversible structural change associated with calcium binding.

  4. Role of electrostatic repulsion on colloidal stability of Bacillus halmapalus alpha-amylase

    DEFF Research Database (Denmark)

    Olsen, Søren Nymand; Andersen, Kim Bruno; Randolf, Theodor

    2009-01-01

    Bacillus halmapalus α-amylase (BHA) as a model protein. Repulsive forces between partly unfolded monomers were shown to strongly affect aggregation. Adding salt, increasing valence of counter ions or decreasing pH in the direction of pI resulted in a shift in the rate-limiting step from association...

  5. 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.

  6. N-terminal amino acid sequence of Bacillus licheniformis alpha-amylase: comparison with Bacillus amyloliquefaciens and Bacillus subtilis Enzymes.

    OpenAIRE

    Kuhn, H; Fietzek, P P; Lampen, J O

    1982-01-01

    The thermostable, liquefying alpha-amylase from Bacillus licheniformis was immunologically cross-reactive with the thermolabile, liquefying alpha-amylase from Bacillus amyloliquefaciens. Their N-terminal amino acid sequences showed extensive homology with each other, but not with the saccharifying alpha-amylases of Bacillus subtilis.

  7. N-terminal amino acid sequence of Bacillus licheniformis alpha-amylase: comparison with Bacillus amyloliquefaciens and Bacillus subtilis Enzymes.

    Science.gov (United States)

    Kuhn, H; Fietzek, P P; Lampen, J O

    1982-01-01

    The thermostable, liquefying alpha-amylase from Bacillus licheniformis was immunologically cross-reactive with the thermolabile, liquefying alpha-amylase from Bacillus amyloliquefaciens. Their N-terminal amino acid sequences showed extensive homology with each other, but not with the saccharifying alpha-amylases of Bacillus subtilis. PMID:6172418

  8. 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.

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

    Science.gov (United States)

    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. PMID:7665495

  10. CHARACTERIZATION OF A NEW BACILLUS-STEAROTHERMOPHILUS ISOLATE - A HIGHLY THERMOSTABLE ALPHA-AMYLASE-PRODUCING STRAIN

    NARCIS (Netherlands)

    WIND, RD; BUITELAAR, RM; EGGINK, G; HUIZING, HJ; DIJKHUIZEN, L

    A novel strain of Bacillus stearothermophilus was isolated from samples of a potato-processing industry. Compared to known alpha-amylases from other B. stearothermophilus strains, the isolate was found to produce a highly thermostable alpha-amylase. The half-time of inactivation of this

  11. 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.

  12. Effect of decoyinine on the regulation of alpha-amylase synthesis in Bacillus subtilis.

    OpenAIRE

    Nicholson, W L; Chambliss, G H

    1987-01-01

    Decoyinine, an inhibitor of GMP synthetase, allows sporulation in Bacillus subtilis to initiate and proceed under otherwise catabolite-repressing conditions. The effect of decoyinine on alpha-amylase synthesis in B. subtilis, an event which exhibits regulatory features resembling sporulation initiation, was examined. Decoyinine did not overcome catabolite repression of alpha-amylase synthesis in a wild-type strain of B. subtilis but did cause premature and enhanced synthesis in a mutant strai...

  13. Spectroscopic study on the interaction of Bacillus subtilis {alpha}-amylase with cetyltrimethylammonium bromide

    Energy Technology Data Exchange (ETDEWEB)

    Omidyan, R., E-mail: r.omidyan@sci.ui.ac.i [Department of Chemistry, University of Isfahan, Isfahan 81746-73441 (Iran, Islamic Republic of); Department of Chemistry, Institute for Advanced Studies in Basic Sciences (IASBS), Zanjan 45137-66731 (Iran, Islamic Republic of); Kazemi, S.H. [Department of Chemistry, Institute for Advanced Studies in Basic Sciences (IASBS), Zanjan 45137-66731 (Iran, Islamic Republic of); Bordbar, A.K. [Department of Chemistry, University of Isfahan, Isfahan 81746-73441 (Iran, Islamic Republic of); Zaynalpour, S. [Department of Chemistry, Institute for Advanced Studies in Basic Sciences (IASBS), Zanjan 45137-66731 (Iran, Islamic Republic of)

    2011-06-15

    The interaction between {alpha}-amylase from Bacillus subtilis and cetyltrimethylammonium bromide (CTAB) has been investigated at various temperature conditions using fluorescence and circular dichroism (CD) spectroscopic methods. Fluorescence data revealed that the fluorescence quenching of {alpha}-amylase by CTAB is the result of complex formation between CTAB and {alpha}-amylase. The thermodynamic analysis on the binding interaction data shows that the interactions are strongly exothermic ({Delta}H{sup o}=-17.92 kJ mol{sup -1}) accompanied with increase in entropy ({Delta}S{sup o} between 109 to 135 J mol{sup -1} K{sup -1}). Thus the binding of CTAB to {alpha}-amylase is both enthalpic and entropic driven, which represent the predominate role of both electrostatic and hydrophobic interactions in complex formation process. The values of 2.17x10{sup -3} M{sup -1} and 1.30 have been obtained from associative binding constant (K{sub a}) and stoichiometry of binding number (n), from analysis of fluorescence data, respectively. Circular dichroism spectra showed the substantial conformational changes in secondary structure of {alpha}-amylase due to binding of CTAB, which represents the complete destruction of both secondary and tertiary structure of {alpha}-amylase by CTAB. - Research highlights: {yields} The Fluorescence quenching effect of {alpha}-amylase by CTAB is a consequence of formation {alpha}-amylase-CTAB complex. {yields} The {alpha}-helical analyzing from the CD spectra in the various concentration of CTAB shows strongly deformation of {alpha}-amylase. {yields} Thermodynamic analysis of quenching verify that the interactions are both enthalpy and entropic driven.

  14. Structure based protein engineering of Bacillus stearothermophilus {alpha}-amylase: toward a new substrate specificity

    Energy Technology Data Exchange (ETDEWEB)

    Rasera, Ana Claudia [Sao Paulo Univ., SP (Brazil). Inst. de Ciencias Biomedicas; Iulek, Jorge [Universidade Estadual de Ponta Grossa, PR (Brazil). Inst. de Quimica; Delboni, Luis Fernando; Barbosa, Valma Martins Barbosa [Parana Univ., Curitiba, PR (Brazil). Dept. de Bioquimica

    1997-12-31

    Full text. Structural similarity is observed in all members of {alpha}-amylase family but different products are generated during hydrolysis of starch due to different affinities for intermediate dextrins. In order to understand the structural determinants for this property and to introduce different specificity to {alpha}-amylase of Bacillus stearothermophilus we intend to solve the three dimensional structure by X-ray crystallography of the native protein by using synchrotron radiation at Brazilian National Synchrotron Light Laboratory (LNLS). Protein was over expressed in E. coli, purified and crystallization experiments were carried out by using sparse matrix Crystal Screen and Crystal Screen II from Hampton Research (Laguna Hills, CA, USA). Several condition have produced crystals with some defined characteristic: MDP seems to be important to the crystallization: the preferential pH is around 7.5 with organic buffer (HEPES); organic solvent as 2-propanol seems to be also important for the crystallization. On those condition crystals appeared as cluster of needles or small crystals with high number of nucleation. New conditions are being prepared to improve the site and quality of crystals. Data collection of native of Bacillus stearothermophilus {alpha}-amylase will e done at Protein Crystallography Station at LNLS. Crystal structure of mutated {alpha}-amylase bu site direct mutagenesis of residues suggested by the native crystal structure will be obtained. Co-crystallization of Bacillus stearothermophilus {alpha}-amylase and oligosaccharide will be carried out to identify residues involved in the binding site to plan new mutation. In another series of mutation the putative binding site residues, once identified, will be mutated to residues observed in TAKA amylase to confer different specificity to {alpha}-amylase. Based on the available TAKA amylase structure, in the primary sequence homology and in the three dimensional model of Bacillus stearothermophilus

  15. 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

  16. Study of the solubility of a modified Bacillus licheniformis alpha-amylase around the isoelectric point

    DEFF Research Database (Denmark)

    Faber, Cornilius; Hobley, Timothy John; Mollerup, Jørgen

    2007-01-01

    The solubility of a modified recombinant Bacillus licheniformis alpha-amylase (mBLA) has been studied by batch crystallization. A semi-pure preparation was chosen containing five isoforms with pI values from 6 to 7.3 (weighted average of 6.6). Small amounts (

  17. Increased production of alpha-amylase by Bacillus amyloliquefaciens in the presence of glycine.

    Science.gov (United States)

    Zhang, Q; Tsukagoshi, N; Miyashiro, S; Udaka, S

    1983-01-01

    The production of alpha-amylase by Bacillus amyloliquefaciens increased by a factor of 300 when glycine was added to a chemically defined simple medium at the early-logarithmic phase of growth. Glycine was not metabolized to a significant extent under the conditions used, but it considerably prevented the lowering of the pH of the culture. PMID:6193759

  18. 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.

  19. 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...

  20. 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.

  1. Expression and secretion of Bacillus amyloliquefaciens alpha-amylase by using the yeast pheromone alpha-factor promoter and leader sequence in Saccharomyces cerevisiae.

    OpenAIRE

    Southgate, V J; Steyn, A J; Pretorius, I S; Van Vuuren, H J

    1993-01-01

    Replacement of the regulatory and secretory signals of the alpha-amylase gene (AMY) from Bacillus amylolique-faciens with the complete yeast pheromone alpha-factor prepro region (MF alpha 1p) resulted in increased levels of extracellular alpha-amylase production in Saccharomyces cerevisiae. However, the removal of the (Glu-Ala)2 peptide from the MF alpha 1 spacer region (Lys-Arg-Glu-Ala-Glu-Ala) yielded decreased levels of extracellular alpha-amylase.

  2. 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.

  3. Purification and characterization of an extracellular alpha-amylase from Bacillus subtilis AX20.

    Science.gov (United States)

    Najafi, Mohsen Fathi; Deobagkar, Dileep; Deobagkar, Deepti

    2005-06-01

    A Bacillus subtilis AX20 from soil with ability to produce extracellular alpha-amylases was isolated. The characterization of microorganism was performed by biochemical tests as well as 16S rDNA sequencing. Maximum amylase activity (38 U/ml) was obtained at stationery phase when the culture was grown at 37 degrees C. The enzyme was purified to homogeneity with an overall recovery of 24.2% and specific activity of 4133 U/mg. The native protein showed a molecular mass of 149 kDa composed of a homodimer of 78 kDa polypeptide by SDS-PAGE. The optimum pH and temperature of the amylase were 6 and 55 degrees C, respectively. The enzyme was inhibited by Hg(2+), Ag(2+), and Cu(2+) and it did not show an obligate requirement of metal ions. The enzyme was not inhibited by EDTA or EGTA, suggesting that this enzyme is not a metalloenzyme. The end products of corn starch and soluble starch were glucose (70-75%) and maltose (20-25%). Rapid reduction of blue value and the end products suggest an endo mode of action for the amylase. The purified amylase shows interesting properties useful for industrial applications.

  4. 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.

  5. Characteristics of raw starch degrading alpha-amylase from Bacillus aquimaris MKSC 6.2 associated with soft coral Sinularia sp.

    NARCIS (Netherlands)

    Puspasari, Fernita; Nurachman, Zeily; Noer, Achmad Saefuddin; Radjasa, Ocky Karna; van der Maarel, Marc J. E. C.; Natalia, Dessy

    Partially purified alpha-amylase from Bacillus aquimaris MKSC 6.2, a bacterium isolated from a soft coral Sinularia sp., Merak Kecil Island, West Java, Indonesia, showed an ability to degrade raw corn, rice, sago, cassava, and potato starches with adsorption percentage in the range of 65-93%. Corn

  6. Effect of gelatinization and hydrolysis conditions on the selectivity of starch hydrolysis with alpha-amylase from Bacillus licheniformis.

    Science.gov (United States)

    Baks, Tim; Bruins, Marieke E; Matser, Ariette M; Janssen, Anja E M; Boom, Remko M

    2008-01-23

    Enzymatic hydrolysis of starch can be used to obtain various valuable hydrolyzates with different compositions. The effects of starch pretreatment, enzyme addition point, and hydrolysis conditions on the hydrolyzate composition and reaction rate during wheat starch hydrolysis with alpha-amylase from Bacillus licheniformis were compared. Suspensions of native starch or starch gelatinized at different conditions either with or without enzyme were hydrolyzed. During hydrolysis, the oligosaccharide concentration, the dextrose equivalent, and the enzyme activity were determined. We found that the hydrolyzate composition was affected by the type of starch pretreatment and the enzyme addition point but that it was just minimally affected by the pressure applied during hydrolysis, as long as gelatinization was complete. The differences between hydrolysis of thermally gelatinized, high-pressure gelatinized, and native starch were explained by considering the granule structure and the specific surface area of the granules. These results show that the hydrolyzate composition can be influenced by choosing different process sequences and conditions.

  7. Concomitant production of two proteases and alpha-amylase by a novel strain of Bacillus subtilis in a microprocessor controlled bioreactor

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    Hamid Mukhtar

    2012-09-01

    Full Text Available We describe the simultaneous production of Bacillus subtilis based proteases and alpha amylase using a computer controlled laboratory scale 7.5 L batch bioreactor. The present strain is the first to be reported that concomitantly produces these two industrially important enzymes. The growth and sporulation of Bacillus subtilis was monitored and maximum production of alkaline protease and alpha amylase was found to coincide with maximum sporulation. Two types of proteases were detected in the fermentation broth; a neutral and an alkaline protease most active in a pH range of 7.0-8.0 and 8.0-10, respectively. Maximum production of proteases was observed at an incubation temperature of 37ºC while that of alpha amylase was observed at 40ºC. The optimum aeration and agitation levels for protease production were 0.6 L/L/min and 200rpm, respectively, and for alpha amylase were 0.6 L/L/min and 150 rpm. The kinetic parameters Yp/x and qp were also found to be significant at the given fermentation conditions.

  8. A statistical approach for the production of thermostable and alklophilic alpha-amylase from Bacillus amyloliquefaciens KCP2 under solid-state fermentation.

    Science.gov (United States)

    Prajapati, Vimal S; Trivedi, Ujjval B; Patel, Kamlesh C

    2015-04-01

    The bacterial strain producing thermostable, alklophilic alpha-amylase was identified as Bacillus amyloliquefaciens KCP2 using 16S rDNA gene sequencing data (NCBI Accession No: KF112071). Medium components were optimized through the statistical approach for the synthesis of alpha-amylase by the organism under solid-state fermentation using wheat bran as the substrate. The medium components influencing the enzyme production were identified using a two-level fractional factorial Plackett-Burman design. Among the various variables screened, starch, ammonium sulphate and calcium chloride were found to be most significant medium components. The optimum levels of these significant parameters were determined employing the response surface Central Composite design which significantly increased the enzyme production with the supplementation of starch 0.01 g, ammonium sulphate 0.2 g and 5 mM calcium chloride in the production medium. Temperature and pH stability of the alpha-amylase suggested its wide application in the food and pharmaceutical industries.

  9. Enzymatic laundry for old clothes: immobilized alpha-amylase from Bacillus sp. for the biocleaning of an ancient Coptic tunic.

    Science.gov (United States)

    Ferrari, Martina; Mazzoli, Roberto; Morales, Simona; Fedi, Mariaelena; Liccioli, Lucia; Piccirillo, Anna; Cavaleri, Tiziana; Oliva, Cinzia; Gallo, Paolo; Borla, Matilde; Cardinali, Michela; Pessione, Enrica

    2017-09-01

    The classification and conservation of ancient artworks (belonging to collections) is of important cultural, historical, and economic concern. However, ancient textiles often display structural damage that renders them fragile and unsuitable for exhibition. One of the most common types of damage is linked to erroneous restoration treatments, among which the application of glues to consolidate cuts. Harsh strategies, such as mechanical or chemical treatments, are not suitable since they can cause further impairment of the fabric, whereas mild approaches, like wet cleaning, are often ineffective, as also demonstrated by the present study. Here, we have explored the possibility of using gellan-immobilized enzymes of bacterial origin (Bacillus alpha-amylase) to obtain a satisfactory starch removal from a damaged archaeological tunic-shroud from the Turin Egyptian Museum (Italy), without altering the original yarns or textile fibers. This method, already applied to clean casein-damaged wall paintings, as well as cotton, silk, and linen fabrics, has proved to be optimal for the treatment of a wool burial shroud and to be able to definitively solve fragile textile restoration problems. Moreover, efforts have been made to obtain insights into the artwork: a multidisciplinary approach has allowed to obtain a correct chronological attribution (radiocarbon dating) and fabric fiber characterization (SEM-EDX) as well as shed light on the colored parts and dark stains (FORS+IRFC and XRF). Finally, the evaluation of the type of glue, by Fourier transform infrared spectroscopy, has suggested the best enzyme for glue removal. These results have demonstrated that a mild bio-based approach is a successful tool for the treatment of archaeological textiles in critical conditions.

  10. Isolation of Alpha-amylase Producing Thermophilic Bacillus Strains and Partial Characterization of the Enzymes

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    Celal Türker

    2015-03-01

    Full Text Available In the present study, we isolated three thermophilic Bacillus strains from the soil samples collected from the coast sediments of the Burnaz Stream located in Erzin. The isolates were entitled as Bacillus sp. CT1, CT2, and CT3, respectively. The maximum α-amylase production was revealed at 60°C for CT1 strain, and at 80°C for CT2 and CT3 strains, respectively. The optimum enzyme activity was observed at 90°C for CT1 α-amylase, whereas at 60°C for CT2 and CT3 α-amylases. On the other hand, optimum pH value for CT2 α-amylase was 7.0, whereas 8.0 for CT1 and CT3 α-amylases. The specific activities of CT1, CT2, and CT3 amylases were 317.6, 113.3 and 362.7 U/mg at 55°C, respectively. The estimated molecular weight of CT1 and CT3 α-amylase was 65 kDa, and for CT2 α-amylase was 38 kDa by zymogram analysis.

  11. 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)

  12. Wheat bran as a substrate for thermo stable alpha-amylase production by gamma irradiated bacillus megaterium in solid state fermentation

    International Nuclear Information System (INIS)

    ElVatal, A.I.; Khalaf, M.A.

    2003-01-01

    Thermo stable alpha-amylase (EC 3.2.1.1) production from cheap agriculture-industrial waste wheat bran (WB) medium by superior potent gamma irradiated locally isolated strain of Bacillus megaterium in solid state fermentation (SSF) was studied. A highly yielding, stable enhanced isolated strain of bacillus megaterium in solid state fermentation (SSF) was studied. A highly yielding stable enhanced isolate B. megaterium- gamma 21F derived from the 10 kGy, treatment, exhibited the highest alpha-amylase activity under SSF, with 2.8 fold more enzyme titer as compared to the unirradiated wild strain. A vancomycin (Vm) resistant gamma irradiated enhanced isolate B. megaterium-gamma 21F2 (which was selected throughout the subsequent work) secreted (1.27 and 3.58) folds superior titers of alpha-amylase than the gamma irradiated parent isolate (B.megaterium -gamma21F) and unirradiated wild strain, respectively under SSF process. The effects of various parameters, such as moistening agent, initial moisture content level, initial ph, incubation temperature, inoculum size and incubation time on thermo stable alpha-amylase production by B.megaterium-gamma 21F2 under SSF were studied. Maximum enzyme production was recorded in WB medium moistened with (1:2, w/v) distilled water at initial ph (7.0) and inoculated with (2.24 x 10 8 cells/g WB) after 48 h incubation at 40 C degree. Between different solvents used for enzyme extraction from fermented WB mass, distilled water at ph (7.0) was the superior efficient leaching solvent. The specific activity of the precipitated partially purified crude thermo stable enzyme was (258.7 U/mg protein) with ph optima (6.5-7.0), at optimal temperatures (65-70 c degree) and it retained about 53% of its maximum activity after 12 h incubation at 70 c degree. The partially purified crude enzyme was used for starch digestion (5%0 under optimized reaction conditions, wherein (98.2%) starch hydrolysis was attained after 6 h

  13. New insights into the effectiveness of alpha-amylase enzyme presentation on the Bacillus subtilis spore surface by adsorption and covalent immobilization.

    Science.gov (United States)

    Gashtasbi, Fatemeh; Ahmadian, Gholamreza; Noghabi, Kambiz Akbari

    2014-10-01

    Most of the studies in the field of enzyme immobilization have sought to develop a simple, efficient and cost-effective immobilization system. In this study, probiotic Bacillus spores were used as a matrix for enzyme immobilization, because of their inherent resistance to extreme temperatures, UV irradiation, solvents and drying. Above all, their preparation is cost-effective. The alpha-amylase enzyme was immobilized on the spore surface by the covalent and adsorption methods. For the covalent method, 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC) and N hydroxysulfosuccinimide (NHS) were used. The maximum concentration of the alpha-amylase immobilized by the two methods onto the spore surface was 360 μg/1.2×10(11) spore. However, maximum activity was achieved at an enzyme concentration of approximately 60 μg/.4×10(10), corresponding to an estimated activity of 8×10(3) IU mg(-1)/1.2×10(11) spore for covalent immobilization and 8.53×10(3) for the adsorption method. After washing the enzyme with 1M NaCl and 0.5% Triton X-100, the enzyme immobilization yield was estimated to be 77% and 20.07% for the covalent and adsorption methods, respectively. The alpha-amylase immobilized by both methods, displayed improved activity in the basic pH range. The optimum pH for the free enzyme was 5 while it shifted to 8 for the immobilized enzyme. The optimum temperatures for the free and immobilized enzymes were 60 °C and 80 °C, respectively. The covalently-immobilized alpha-amylase retained 65% of its initial activity, even after 10 times of recycling. The Km and Vmax values were determined by the GraphPad Prism software, which showed that the Vmax value decreased moderately after immobilization. This is the first study which reports the covalent immobilization of an enzyme on the spore surface. Copyright © 2014 Elsevier Inc. All rights reserved.

  14. Solid State Fermentation of a Raw Starch Digesting Alkaline Alpha-Amylase from Bacillus licheniformis RT7PE1 and Its Characteristics

    Directory of Open Access Journals (Sweden)

    Romana Tabassum

    2014-01-01

    Full Text Available The thermodynamic and kinetic properties of solids state raw starch digesting alpha amylase from newly isolated Bacillus licheniformis RT7PE1 strain were studied. The kinetic values Qp, Yp/s, Yp/X, and qp were proved to be best with 15% wheat bran. The molecular weight of purified enzyme was 112 kDa. The apparent Km and Vmax values for starch were 3.4 mg mL−1 and 19.5 IU mg−1 protein, respectively. The optimum temperature and pH for α-amylase were 55°C, 9.8. The half-life of enzyme at 95°C was 17h. The activation and denaturation activation energies were 45.2 and 41.2 kJ mol−1, respectively. Both enthalpies (ΔH∗ and entropies of activation (ΔS∗ for denaturation of α-amylase were lower than those reported for other thermostable α-amylases.

  15. 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...

  16. Synergistic Effects of Agro-Industrial Wastes on Alpha Amylase Production by Bacillus amyloliquefaciens in Semi Solid Substrate Fermentation

    Directory of Open Access Journals (Sweden)

    Ali Yaraş

    2015-12-01

    Full Text Available This study concerns with the combinations of soybean meal (SM, wheat bran (WB and whey were utilized as the substrates for α-amylase production by Bacillus amyloliquefaciens NRRL B-645. As a result of the experiments, the highest α-amylase activity (4257 U/ml was obtained containing SM 20 g/l, WB 5 g/l, whey 5% (v/v, peptone 1 g/l, yeast extract (YE 0.5 g/l and (NH42SO4 1 g/l at 33 °C and 150 rpm for 48 h. This innovative process for the α-amylase production can be extended to different enzymes using various agro-industrial wastes.

  17. Enzymatic Properties of an Alkaline and Chelator Resistant alpha-amylase from the Alkaliphilic Bacillus sp. Isolate L1711

    Science.gov (United States)

    Bernhardsdotter, Eva C. M. J.; Pusey, Marc L.; Ng, Joseph D.; Garriott, Owen K.

    2004-01-01

    An alkaliphilic amylase producing bacterium, Bacillus sp. strain L 711, was selected among 13 soda lakes isolates. When grown at pH 10.5 and 37 C, strain L711 produced multiple forms of amylases in the culture broth. One of these, BAA, was purified from the culture supernatant by QAE column chromatography and preparative native gel electrophoresis. The molecular weight of BAA was determined to be 51 kDa by denaturing gel electrophoresis. The pH optima for activity below and above 40 C were 9.5 - 10.0 and 7.0 - 7.5 respectively. BAA was stable in the pH range 6-11 and was completely inactivated at 55 C. The thermostability was not increased in the presence of Ca(2+). The enzyme was strongly inhibited by Ca(2+), Zn(2+), Mg(2+), Mn(2+), Ba(2+) and Cu(2+), whereas the presence of Na(+), Co(2+) and EDTA (10 mM) enhanced enzymatic activity. The K(sub m), and specific activity of BAA on soluble starch were 1.9 mg/ml and 18.5 U/mg respectively. The main end products of hydrolysis were maltotetraose, maltose and glucose.

  18. Production and Partial Characterization of Alpha-Amylase from Corn ...

    African Journals Online (AJOL)

    Corn pomace, a by-product of corn from “ogi” production was used as a substrate for the production of alpha-amylase by a corn fermenting strain of Bacillus licheniformis ATCC 4527. The corn pomace was fortified with potassium phosphate, magnesium sulphate and 2% w/v glucose to produce corn pomace basal medium, ...

  19. Significance of HPr in catabolite repression of alpha-amylase.

    Science.gov (United States)

    Voskuil, M I; Chambliss, G H

    1996-01-01

    CcpA and HPr are presently the only two proteins implicated in Bacillus subtilis global carbon source catabolite repression, and the ptsH1 mutation in the gene for the HPr protein was reported to relieve catabolite repression of several genes. However, alpha-amylase synthesis by B. subtilis SA003 containing the ptsH1 mutation was repressed by glucose. Our results suggest HPr(Ser-P) may be involved in but is not required for catabolite repression of alpha-amylase, indicating that HPr(Ser-P) is not the sole signaling molecule for CcpA-mediated catabolite repression in B. subtilis. PMID:8955329

  20. Chemical synthesis of a dual branched malto-decaose: A potential substrate for alpha-amylases

    DEFF Research Database (Denmark)

    Damager, Iben; Jensen, Morten; Olsen, Carl Erik

    2005-01-01

    A convergent block strategy for general use in efficient synthesis of complex alpha-(1 -> 4)- and alpha-(1 -> 6)-malto-oligosaccharides is demonstrated with the first chemical synthesis of a malto-oligosaccharide, the decasoccharide 6,6""-bis(alpha-maltosyl)-maltohexaose, with two branch points....... Using this chemically defined branched oligosaccharide as a substrate, the cleavage pattern of seven different alpha-amylases were investigated. alpha-Amylases from human saliva, porcine pancreas, barley alpha-amylose 2 and recombinant barley alpha-amylase 1 all hydrolysed the decasaccharide selectively....... This resulted in a branched hexasaccharide and a branched tetrasoccharide. alpha-Amylases from Asperagillus oryzae, Bacillus licheniformis and Bacillus sp. cleaved the decasoccharide at two distinct sites, either producing two branched pentasoccharides, or a branched hexasoccharide and a branched...

  1. Production of Cellulases, Xylanase, Pectinase, alpha-amylase and Protease Enzymes Cocktail by Bacillus spp. and Their Mixed Cultures with Candida tropicalis and Rhodotorula glutinis under Solid State Fermentation

    International Nuclear Information System (INIS)

    El-Batal, A.I.; Abo-State, M.A.

    2006-01-01

    A group of twelve locally isolated Bacillus species, B.megaterium (MAI and MA II), B.licheniformis (MLI and ML II); B. circulans, B. stearothermophilis, B.cereus, B.sphaericus, B. pumilus, B. laterosporus, B. coagulans and B. pantothenticus, were examined for the production of cellulases, xylanase, pectinase, alpha-amylase and protease enzymes cocktail on wheat bran under solid state fermentation (SSF). All species were found to be potent hydrolyzing enzymes producers and the superior producing species were B. megaterium MAI and B. licheniformis. On the other hand, both of them still produced highest enzyme titres when mixed with Candida tropicalis or Rhodotorula glutinis, yeast strains. The two superior bacterial strains produced the highest enzymatic activities when coculturing with C. tropicalis compared with coculturing with R. glutinis only or with both C. tropicalis and R. glutinis in combination. The inferior activities of cocultures (B. megaterinm MAI and R. glutinis) were enhanced in carboxymethyl cellulase (CMCase), filter paper cellulase (FPase), avecilase, xylanase, pectinase, -amylase and protease by gamma irradiation at dose 1.0 kGy with percent increase 8 %, 20 %, 10 %, 4 %, 31 %, 22 % and 34 %, respectively as compared with un-irradiated cocultures

  2. Production and properties of alpha-amylase from thermophilic Bacillus sp. Produção e propriedades de alfa-amilase de Bacillus sp. termofílico

    Directory of Open Access Journals (Sweden)

    Carlos Alberto Martins Cordeiro

    2002-01-01

    Full Text Available alpha-amylase (1,4-alpha-D-glucan glucanohydrolase, EC 3.2.1.1 production by thermophilic Bacillus sp strain SMIA-2 cultivated in liquid media containing soluble starch reached a maximum at 48h, with levels of 57U/mL. Studies on the a-amylase characterization revealed that the optimum temperature for activity was 70ºC. The enzyme was stable for 2h at 50ºC, while at 60ºC, 70ºC and 90ºC, 4%, 13% and 38% of the original activities were lost, respectively. The optimum pH of the enzyme was 7.5. After incubation of crude enzyme solution for 24h at pH 7.5, a decrease of about 5% of its original activity was observed. The enzyme was strongly inhibited by Co2+, Cu2+ and Ba2+, but less affected by Ca2+, Mg2+, Ni2+, Sr2+ and Mn2+. The enzyme in 1M and 5M NaCl solutions the enzyme retained 70% and 47% of the original activity after 24h of incubation at 4ºC, respectively.A produção de alfa-amilase (1,4-alfa-D-glicano glicanohidrolase, EC 3.2.1.1 por um Bacillus sp cepa SMIA-2 cultivado em meios líquidos contendo amido solúvel, alcançou o máximo em 48h com níveis de 57U/mL. Estudos sobre a caracterização de alfa-amilase revelaram que a temperatura ótima de atividade desta enzima foi 70ºC. A enzima foi estável por 2h a 50ºC, enquanto que a 60ºC, 70ºC e 90ºC, 4%, 13% e 38% da atividade original foram perdidas, respectivamente. O pH ótimo da enzima foi 7,5. Após a incubação da enzima bruta por 24h a pH 7,5 observou-se um decréscimo em torno de 5% de sua atividade original. A enzima foi fortemente inibida por Co2+, Cu2+ e Ba2+, mas foi menos afetada por Ca2+, Mg2+, Ni2+, Sr2+ e Mn2+. Em solução de NaCl 1M e 5M, a enzima reteve 70% e 47% da sua atividade original após 24h a 4ºC, respectivamente.

  3. Raw starch-degrading alpha-amylase from Bacillus aquimaris MKSC 6.2 : isolation and expression of the gene, bioinformatics and biochemical characterization of the recombinant enzyme

    NARCIS (Netherlands)

    Puspasari, F.; Radjasa, O. K.; Noer, A. S.; Nurachman, Z.; Syah, Y. M.; van der Maarel, M.; Dijkhuizen, L.; Janecek, S.; Natalia, D.; Janeček, Š.

    Aims The aims were to isolate a raw starchdegrading a-amylase gene baqA from Bacillus aquimaris MKSC 6.2, and to characterize the gene product through in silico study and its expression in Escherichia coli. Methods and Results A 1539 complete open reading frame of a starchdegrading a-amylase gene

  4. Studies on alkali-modified cassava starch - changes of structural and enzyme (. alpha. -amylase) susceptibility properties

    Energy Technology Data Exchange (ETDEWEB)

    Raja, K.C.M. (Regional Research Lab., Trivandrum (India). Fermentation Section)

    1992-04-01

    Properties of cassava starch could be modified by subjecting to alkali treatment under controlled experimental conditions. Modified starch samples showed lower amylose content and higher alkali number. Compared to untreated starch samples, alkali modified starches had higher {alpha}-amylase (Bacillus sp.) susceptibility. The properties could be advantageously made use of for preparing maltodextrins having DE 20-23. (orig.).

  5. Enzymatic Properties of an Alkaline and Chelator Resistant Proportional to alpha-Amylase from the Alkaliphilic Bacillus sp. Isolate L1711

    Science.gov (United States)

    Bernhardsdotter, Eva C. M. J.; Pusey, Marc L.; Ng, Joseph D.; Garriott, Owen K.

    2004-01-01

    An alkaliphilic amylase producing bacterium, Bacillus sp. strain L1711, was selected among 13 soda lakes isolates. When grown at pH 10.5 and 370 C, strain L1711 produced multiple forms of amylases in the culture broth. One of these, BAA, was purified from the culture supernatant by QAE column chromatography and preparative native gel electrophoresis. The molecular weight of BAA was determined to be 51 kDa by denaturing gel electrophoresis. The pH optima for activity below and above 40 C were 9.5-10.0 and 7.0-7.5 respectively. BAA was stable in the pH range 6-11 and was completely inactivated at 55?C. The thermostability was not increased in the presence of Ca(2+). The enzyme was strongly inhibited by Ca(2+), Zn(2+), Mg(2+), Mn(2+), Ba(2+) and Cu(2+), whereas the presence of Na(+), Co2+ and EDTA (10 mM) enhanced enzymatic activity. The K(sub m) and specific activity of BAA on soluble starch were 1.9 mg/ml and 18.5 U/mg respectively. The main end products of hydrolysis were maltotetraose, maltose and glucose .

  6. Purification and biochemical characterization of a thermostable and acid-stable alpha-amylase from Bacillus licheniformis B4-423.

    Science.gov (United States)

    Wu, Xiangrong; Wang, Yuxia; Tong, Bending; Chen, Xianghua; Chen, Jianhua

    2018-04-01

    Novel thermostable amylase need to be continuously explored with the improvement of industrial requirements. A new acidophilic and thermostable amylase producing bacterium isolated from spring was identified as Bacillus strain on the basis of 16S rDNA. The amylase was purified by ammonium sulphate precipitation, gel chromatography and anion exchange chromatography. SDS-PAGE revealed that the enzyme was monomeric with a molecular weight of 58 kDa. The amylase exhibited optimal activity at pH 5.0 and temperature 100 °C. Then the enzyme showed high stability in pH ranges 4.0-10.0 and more than 90% of maximal activity was found from 20 °C to 80 °C. Apart from good stability toward SDS and non-ionic detergent, the purified enzyme exhibited high compatibility with some inhibitors such as urea and EDTA. The results demonstrated the stability of the enzyme in different organic solvents. Moreover, we determined the amylase gene, compared the structure with α-amylase BAA and BLA and found some thermostability determinants in our enzyme. Overall, presenting various properties were including high thermostability, Ca 2+ -independency, broad temperature and pH profiles, organic-solvent tolerance as well as excellent stability with detergents. Such characteristics have not been reported for this type of enzyme, and the α-amylase will be a suitable candidate in industrial fields. Copyright © 2017 Elsevier B.V. All rights reserved.

  7. 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.

  8. Purification and characterization of alpha-amylase from rat pancreatic acinar carcinoma. Comparison with pancreatic alpha-amylase.

    OpenAIRE

    Reddy, M K; Heda, G D; Reddy, J K

    1987-01-01

    alpha-Amylase was purified to apparent homogeneity from normal pancreas and a transplantable pancreatic acinar carcinoma of the rat by affinity chromatography on alpha-glucohydrolase inhibitor (alpha-GHI) bound to aminohexyl-Sepharose 4B. Recovery was 95-100% for both pancreas and tumour alpha-amylases. They were monomeric proteins, with Mr approx. 54000 on SDS/polyacrylamide-gel electrophoresis. Isoelectric focusing of both normal and tumour alpha-amylases resolved each into two major isoenz...

  9. Measurement of microbial alpha-amylases with p-nitrophenyl glycosides as the substrate complex.

    OpenAIRE

    Trepeta, R W; Edberg, S C

    1984-01-01

    The detection of alpha-amylase is commonly used in clinical microbiology laboratories to aid in differentiating Streptococcus bovis from other streptococci. It is also useful in identifying Eikenella corrodens and the gravis subspecies of Corynebacterium diphtheriae and in separating species of the genera Bacteroides, Clostridium, Actinomyces, and Bacillus. Currently, the most frequently used procedure utilizes starch as the substrate and iodine as the indicator. Starch is incorporated into a...

  10. Alpha-amylase inhibition kinetics by caulerpenyne

    Directory of Open Access Journals (Sweden)

    S. CENGIZ

    2010-03-01

    Full Text Available Many algae have important secretions which are generally used for defensive purposes. These secretions take attentions of a lot of researchers who are wondering if these metabolites can be used for medical researches or not. Among these metabolites, caulerpenyne (CYN which is the main metabolite of Caulerpa species, have had an important place in Caulerpa researches since the results related to its determined properties such as cytotoxic, antiviral, antiproliferative and apoptotic effects have been proven by many scientific reports. In the present study, the inhibitory effect of CYN isolated from C. prolifera on alpha-amylase was investigated. The inhibition experiments were done with CYN by spectrophotometric determination method. In order to evaluate the type of inhibition Lineweaver–Burk plot was produced. The results obtained from enzyme kinetic studies exhibited an un-competitive type of inhibition, which is characterized by the difference of Vmax and KM from those of the free enzyme, of alpha-amylase in the presence of CYN. The present study showed that Caulerpa species can be a potential target for producing diabetic drugs in the light of the results obtained for CYN.

  11. 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.

  12. 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...

  13. 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....

  14. Structural basis of alpha-amylase activation by chloride

    OpenAIRE

    Aghajari, N.; Feller, Georges; Gerday, Charles; Haser, R.

    2002-01-01

    To further investigate the mechanism and function of allosteric activation by chloride in some alpha-amylases, the structure of the bacterial alpha-amylase from the psychrophilic micro-organism Pseudoalteromonas haloplanktis in complex with nitrate has been solved at 2.1 Angstrom, as well as the structure of the mutants Lys300Gln (2.5 Angstrom) and Lys300Arg (2.25 Angstrom). Nitrate binds strongly to alpha-amylase but is a weak activator. Mutation of the critical chloride ligand Lys300 into G...

  15. Effect of polyols on fungal alpha-amylase thermostability

    OpenAIRE

    Graber, Marianne; Combes, Didier

    1989-01-01

    International audience; The influence of different polyols (ethylene glycol, glycerol, erythritol, xylitol, and sorbitol) on the thermostability of fungal alpha-amylase at 60°C has been studied. The results obtained show a stabilizing effect in the presence of polyols. In the case of 4 m sorbitol solution, the enzyme half-life is 2000-fold longer than in pure water. These polyols have been found as competitive inhibitors for alpha-amylase andtheir stabilizing effect has been correlated to the...

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

    Energy Technology Data Exchange (ETDEWEB)

    Scannapieco, F.A.; Bergey, E.J.; Reddy, M.S.; Levine, M.J. (State Univ. of New York, Buffalo (USA))

    1989-09-01

    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 {sup 125}I-labeled HSMSL or {sup 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 ({sup 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.

  17. 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

  18. 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

  19. THE INFLUENCE OF ALPHA AMYLASE ON THE QUALITY OF BREAD

    OpenAIRE

    RODICA CHEREJI; IULIANA CRETESCU; RODICA CĂPRIłĂ

    2008-01-01

    This study determined the quality of bread obtained from the control sample flour (M) and the quality of bread obtained from flour with addition of 3 different percentages of alpha amylase (P1-280000 U.SKB/ 100kg flour; P2-560000 U.SKB/ 100kg flour;P3-840000 U.SKB/ 100kg flour). Fungal alpha amylase was used in these concentrations in order to establish which one is the most suitable to be added in flour in order to obtain superior quality characteristics for bread: superior volume of bread, ...

  20. 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...

  1. SYNTHESIS AND PROCESSING OF ESCHERICHIA-COLI TEM-BETA-LACTAMASE AND BACILLUS-LICHENIFORMIS ALPHA-AMYLASE IN ESCHERICHIA-COLI : THE ROLE OF SIGNAL PEPTIDASE-I

    NARCIS (Netherlands)

    van Dijl, J M; SMITH, H; BRON, S; VENEMA, G

    A mutant of Escherichia coli, in which signal peptidase I synthesis can be regulated, was constructed. The mutant was used to study the effects of signal peptidase I limitation on the synthesis and efficiency of processing of two proteins: the periplasmic E. coli TEM-beta-lactamase and Bacillus

  2. 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.

  3. 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

  4. 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....

  5. THE INFLUENCE OF ALPHA AMYLASE ON THE QUALITY OF BREAD

    Directory of Open Access Journals (Sweden)

    RODICA CHEREJI

    2008-05-01

    Full Text Available This study determined the quality of bread obtained from the control sample flour (M and the quality of bread obtained from flour with addition of 3 different percentages of alpha amylase (P1-280000 U.SKB/ 100kg flour; P2-560000 U.SKB/ 100kg flour;P3-840000 U.SKB/ 100kg flour. Fungal alpha amylase was used in these concentrations in order to establish which one is the most suitable to be added in flour in order to obtain superior quality characteristics for bread: superior volume of bread, finer texture of the bread, prolonging the freshness of the bread, improving the color and flavor of the bread, improving the slicing proprieties of the bread.

  6. Salivary Cortisol, Salivary Alpha Amylase, and the Dental Anxiety Scale

    OpenAIRE

    Sadi, Hana; Finkelman, Matthew; Rosenberg, Morton

    2013-01-01

    The aim of this study was to investigate the correlation between dental anxiety, salivary cortisol, and salivary alpha amylase (sAA) levels. Furthermore, the aim was to look into individual differences such as age, race, gender, any existing pain, or traumatic dental experience and their effect on dental anxiety. This study followed a cross-sectional design and included a convenience sample of 46. Every patient was asked to complete the Dental Anxiety Scale (DAS) and a basic demographic/denta...

  7. 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.

  8. Spatio-temporal profiling and degradation of alpha-amylase isozymes during barley seed germination

    DEFF Research Database (Denmark)

    Bak-Jensen, K.S.; Laugesen, Sabrina; Østergaard, Ole

    2007-01-01

    Ten genes from two multigene families encode barley alpha-amylases. To gain insight into the occurrence and fate of individual isoforms during seed germination, the alpha-amylase repertoire was mapped by using a proteomics approach consisting of 2D gel electrophoresis, western blotting, and mass...... spectrometry. Mass spectrometric analysis confirmed that the 29 alpha-amylase positive 2D gel spots contained products of one ( GenBank accession gi| 113765) and two ( gi vertical bar 4699831 and gi vertical bar 166985) genes encoding alpha-amylase 1 and 2, respectively, but lacked products from seven other...... genes. Eleven spots were identified only by immunostaining. Mass spectrometry identified 12 full-length forms and 12 fragments from the cultivar Barke. Products of both alpha-amylase 2 entries co-migrated in five full-length and one fragment spot. The alpha-amylase abundance and the number of fragments...

  9. Clinical and immunological responses to occupational exposure to alpha-amylase in the baking industry.

    OpenAIRE

    Brisman, J; Belin, L

    1991-01-01

    alpha-Amylase is a starch cleaving enzyme often used in the baking industry as a flour additive. It is usually of fungal origin, produced by Aspergillus oryzae. One previous report has shown IgE antibodies and positive skin prick test against alpha-amylase in asthmatic bakers. This paper describes four alpha-amylase sensitised index cases with occupational asthma or rhinitis and the results of a cross sectional study of 20 workers from the same factory who were also exposed to alpha-amylase p...

  10. Salivary Alpha-Amylase Reactivity in Breast Cancer Survivors

    Directory of Open Access Journals (Sweden)

    Cynthia Wan

    2016-03-01

    Full Text Available The two main components of the stress system are the hypothalamic-pituitary-adrenal (HPA and sympathetic-adrenal-medullary (SAM axes. While cortisol has been commonly used as a biomarker of HPA functioning, much less attention has been paid to the role of the SAM in this context. Studies have shown that long-term breast cancer survivors display abnormal reactive cortisol patterns, suggesting a dysregulation of their HPA axis. To fully understand the integrity of the stress response in this population, this paper explored the diurnal and acute alpha-amylase profiles of 22 breast cancer survivors and 26 women with no history of cancer. Results revealed that breast cancer survivors displayed identical but elevated patterns of alpha-amylase concentrations in both diurnal and acute profiles relative to that of healthy women, F (1, 39 = 17.95, p < 0.001 and F (1, 37 = 7.29, p = 0.010, respectively. The average area under the curve for the diurnal and reactive profiles was 631.54 ± 66.94 SEM and 1238.78 ± 111.84 SEM, respectively. This is in sharp contrast to their cortisol results, which showed normal diurnal and blunted acute patterns. The complexity of the stress system necessitates further investigation to understand the synergistic relationship of the HPA and SAM axes.

  11. Cloning of a yeast alpha-amylase promoter and its regulated heterologous expression

    Science.gov (United States)

    Gao, Johnway [Richland, WA; Skeen, Rodney S [Pendleton, OR; Hooker, Brian S [Kennewick, WA; Anderson, Daniel B [Pasco, WA

    2003-04-01

    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.

  12. Morphological characterization of recombinant strains of Aspergillus oryzae producing alpha-amylase during batch cultivations

    DEFF Research Database (Denmark)

    Spohr, Anders Bendsen; Carlsen, Morten; Nielsen, Jens Bredal

    1997-01-01

    Three alpha-amylase producing strains of Aspergillus oryzae used for recombinant protein production have been studied with respect to growth and protein production. By comparing the three strains with respect to morphology and protein production it is shown that a morphological mutant with a more...... dense mycelium is more efficient in producing alpha-amylase....

  13. Binding of carbohydrates and protein inhibitors to the surface of alpha-amylases

    DEFF Research Database (Denmark)

    Bozonnet, Sophie; Bønsager, Birgit Christine; Kramhoft, B.

    2005-01-01

    This review on barley alpha-amylases 1 (AMY1) and 2 (AMY2) addresses rational mutations at distal subsites to the catalytic site, polysaccharide hydrolysis, and interactions with proteinaceous inhibitors. Subsite mapping of barley alpha-amylases revealed 6 glycone and 4 aglycone substrate subsites...

  14. Identification of a novel bean alpha-amylase inhibitor with chitinolytic activity.

    Science.gov (United States)

    Dayler, Charles S A; Mendes, Paulo A M; Prates, Maura V; Bloch, Carlos; Franco, Octavio L; Grossi-de-Sá, Maria F

    2005-10-24

    Zabrotes subfasciatus is a devastating starch-dependent storage bean pest. In this study, we attempted to identify novel alpha-amylase inhibitors from wild bean seeds, with efficiency toward pest alpha-amylases. An inhibitor named Phaseolus vulgaris chitinolytic alpha-amylase inhibitor (PvCAI) was purified and mass spectrometry analyses showed a protein with 33330 Da with the ability to form dimers. Purified PvCAI showed significant inhibitory activity against larval Z. subfasciatus alpha-amylases with no activity against mammalian enzymes. N-terminal sequence analyses showed an unexpected high identity to plant chitinases from the glycoside hydrolase family 18. Furthermore, their chitinolytic activity was also detected. Our data provides compelling evidence that PvCAI also possessed chitinolytic activity, indicating the emergence of a novel alpha-amylase inhibitor class.

  15. Influence of concentration of fragrances on salivary alpha-amylase.

    Science.gov (United States)

    Yamaguchi, M; Tahara, Y; Kosaka, S

    2009-10-01

    The objective is to reveal the influence of the concentration of fragrances on salivary biomarkers, which reflect the human stress system, in 15 female young healthy adults. Lavandula officinalis and Citrus aurantium were used as the test samples. Salivary biomarkers such as alpha-amylase activity (AMY), cortisol (CORT) and dehydroepiandrosterone (DHEA) were measured during baseline, inhalation and post-inhalation periods. Our results indicated that (i) a significant difference was not observed for the control and the 3 wt% test samples, however, the AMY was decreased by inhalation of the 1 wt% test samples (P < 0.05); (ii) AMY levels changed more significantly than did the hormone levels; (iii) a tendency of negative correlation was not observed between DHEA and CORT. It was considered that the time-course change of AMY might be a useful index of the inhalation of fragrances, which reflects the acute psychosomatic reactivity of humans.

  16. Measuring Salivary Alpha-Amylase in the Undergraduate Neuroscience Laboratory.

    Science.gov (United States)

    Bañuelos, Maria S; Musleh, Aya; Olson, Lisa E

    2017-01-01

    Undergraduate courses in biopsychology, neuroscience, and physiology often include laboratory exercises that examine responses to stimulation of the sympathetic nervous system with measurements of heart rate, blood pressure, or galvanic skin levels (sweat response). A newer bioindicator of the sympathetic nervous system is salivary alpha-amylase (sAA) measured with a colorimetic enzyme assay. Undergraduate students successfully measured a rise in sAA due to the stress of giving a class presentation (n=13). Students were enthusiastic to measure a physiological response to a real-life anxiety-producing situation. We describe potential difficulties in the assay and our adaptations to the manufacturer's protocol to make it more feasible in the undergraduate setting.

  17. THE INFLUENCE OF ALPHA AMYLASE ON THE QUALITY OF BREAD

    Directory of Open Access Journals (Sweden)

    CAPRITA RODICA

    2008-01-01

    Full Text Available This study determined the quality of bread obtained from the control sample flour (M and the qualityof bread obtained from flour with addition of 3 different percentages of alpha amylase (P1-280000U.SKB/ 100kg flour; P2-560000 U.SKB/ 100kg flour;P3-840000 U.SKB/ 100kg flour. Fungal alphaamylase was used in these concentrations in order to establish which one is the most suitable to beadded in flour in order to obtain superior quality characteristics for bread: superior volume of bread,finer texture of the bread, prolonging the freshness of the bread, improving the color and flavor of thebread, improving the slicing proprieties of the bread.

  18. The influence of nitrogen sources on the alpha-amylase productivity of Aspergillus oryzae in continuous cultures

    DEFF Research Database (Denmark)

    Pedersen, Henrik; Nielsen, Jens

    2000-01-01

    The influence of the nitrogen source on the cc-amylase productivity of Aspergillus oryzae was quantified in continuous cultivations. Both inorganic and complex nitrogen sources were investigated and glucose was used as the carbon and energy sources. For production of alpha-amylase, nitrate...... in the cc-amylase productivity. The higher alpha-amylase productivity during growth on casein hydrolysate was not caused by increased transcription of the alpha-amylase genes but was caused by a faster secretion of alpha-amylase or by a lower binding of alpha-amylase to the biomass....

  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. Chemical synthesis of a dual branched malto-decaose: A potential substrate for alpha-amylases

    DEFF Research Database (Denmark)

    Damager, Iben; Jensen, Morten; Olsen, Carl Erik

    2005-01-01

    A convergent block strategy for general use in efficient synthesis of complex alpha-(1 -> 4)- and alpha-(1 -> 6)-malto-oligosaccharides is demonstrated with the first chemical synthesis of a malto-oligosaccharide, the decasoccharide 6,6""-bis(alpha-maltosyl)-maltohexaose, with two branch points....... Using this chemically defined branched oligosaccharide as a substrate, the cleavage pattern of seven different alpha-amylases were investigated. alpha-Amylases from human saliva, porcine pancreas, barley alpha-amylose 2 and recombinant barley alpha-amylase 1 all hydrolysed the decasaccharide selectively...

  1. Production of fungal alpha-amylase by Saccharomyces kluyveri in glucose-limited cultivations

    DEFF Research Database (Denmark)

    Møller, Kasper; Sharif, M.Z.; Olsson, Lisbeth

    2004-01-01

    and S. cerevisiae were transformed with the same plasmid, which led to secretion of active alpha-amylase in both cases. The S. cerevisiae 2 mu plasmid was found to be stable in S. kluyveri as evaluated by a constant alpha-amylase productivity in a continuous cultivation for more than 40 generations. S......Heterologous protein production by the yeast Saccharomyces kluyveri was investigated under aerobic glucose-limited conditions. alpha-Amylase from Aspergillus oryzae was used as model protein and the gene was expressed from a S. cerevisiae 2 mu plasmid. For comparison, strains of both S. kluyveri....... kluyveri and S. cerevisiae secreted alpha-amylase with similar yields during continuous cultivations at dilution rates of 0.1 and 0.2 h(-1) (4.8-5.7 mg (g dry weight)(-1)). At a dilution rate of 0.3 h(-1) the metabolism of S. kluyveri was fully respiratory, whereas S. cerevisiae produced significant...

  2. Production of fungal alpha-amylase by Saccharomyces kluyveri in glucose-limited cultivations

    DEFF Research Database (Denmark)

    Møller, Kasper; Sharif, M.Z.; Olsson, Lisbeth

    2004-01-01

    Heterologous protein production by the yeast Saccharomyces kluyveri was investigated under aerobic glucose-limited conditions. alpha-Amylase from Aspergillus oryzae was used as model protein and the gene was expressed from a S. cerevisiae 2 mu plasmid. For comparison, strains of both S. kluyveri...... and S. cerevisiae were transformed with the same plasmid, which led to secretion of active alpha-amylase in both cases. The S. cerevisiae 2 mu plasmid was found to be stable in S. kluyveri as evaluated by a constant alpha-amylase productivity in a continuous cultivation for more than 40 generations. S....... kluyveri and S. cerevisiae secreted alpha-amylase with similar yields during continuous cultivations at dilution rates of 0.1 and 0.2 h(-1) (4.8-5.7 mg (g dry weight)(-1)). At a dilution rate of 0.3 h(-1) the metabolism of S. kluyveri was fully respiratory, whereas S. cerevisiae produced significant...

  3. 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)

  4. Morphology and physiology of an alpha Amylase producing strain of Aspergillus oryzae during batch cultivations

    DEFF Research Database (Denmark)

    Carlsen, Morten; Spohr, Anders Bendsen; Nielsen, Jens Bredal

    1996-01-01

    , whereas the alpha-amylase production has a sharper maximum at about pH 6. During batch cultivation with pellets the growth is described well by the cube-root law when pellet fragmentation can be neglected. The kinetic parameter k in the cube-root law is derived from the growth kinetics with no mass......, suggesting that ethanol is produced in the oxygen limited part of the biomass. A constitutive, low alpha-amylase production was observed at high glucose concentration. The specific alpha-amylase production was significantly higher for filamentous growth than for pellets and oxygen appears to be necessary...... for production of alpha-amylase. (C) 1996 John Wiley & Sons, Inc....

  5. 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...

  6. Ability to bind salivary alpha-amylase discriminates certain viridans group streptococcal species.

    OpenAIRE

    Kilian, M; Nyvad, B

    1990-01-01

    A collection of 144 viridans group streptococcal strains recently characterized as part of a taxonomic study was examined for the ability to bind salivary alpha-amylase. This property was found in most strains of Streptococcus gordonii and Streptococcus mitis and in occasional strains of Streptococcus anginosus and Streptococcus salivarius. In contrast, all strains of Streptococcus sanguis, Streptococcus oralis, Streptococcus vestibularis, and Streptococcus mutans lacked alpha-amylase-binding...

  7. Patterns of cortisol and alpha-amylase reactivity to psychosocial stress in maltreated women.

    Science.gov (United States)

    Mielock, Alyssa S; Morris, Matthew C; Rao, Uma

    2017-02-01

    Childhood maltreatment can trigger enduring changes in major stress response systems, particularly in the context of major depressive disorder (MDD). However, the relative impact of maltreatment versus MDD on hypothalamic-pituitary-adrenal axis and sympathetic-adrenal-medullary system stress reactivity is not well understood. This study examined salivary cortisol and alpha-amylase responses to the Trier Social Stress Test (TSST) in 26 maltreated (15 with current MDD) and 26 non-maltreated (17 with current MDD) women. Maltreated women showed greater anticipatory cortisol reactivity during the TSST protocol compared to non-maltreated women. Maltreated women also showed rapid deceleration in cortisol levels. Whereas non-maltreated women showed initial declines in alpha-amylase levels but rapidly increasing alpha-amylase levels during the TSST protocol, maltreated women did not exhibit changes in alpha-amylase levels during the TSST protocol. Contrary to expectation, MDD did not impact cortisol or alpha-amylase responses. The present study is limited by retrospective report of childhood maltreatment, cross-sectional design, and modest sample sizes. These findings suggest that childhood maltreatment plays a greater role driving alterations in cortisol and alpha-amylase stress reactivity than MDD. Understanding the biological embedding of maltreatment is critical for elucidating mechanisms linking these experiences to risk for negative mental and physical health outcomes. Copyright © 2016 Elsevier B.V. All rights reserved.

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

    Directory of Open Access Journals (Sweden)

    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.

  9. Psychological stress-induced changes in salivary alpha-amylase and adrenergic activity.

    Science.gov (United States)

    Kang, Younhee

    2010-12-01

    The aim of the study was to examine the relationships among salivary alpha-amylase, plasma catecholamines, blood pressure, and heart rate during psychological stress. This study used a pretest-post-test experimental design with a control group, using repeated measures. A total of 33 participants was divided into the experimental group (n = 16) that underwent a college academic final test as the psychological stress and the control group (n = 17) that did not undergo the test. The levels of salivary alpha-amylase and plasma catecholamines, blood pressure, and heart rate were measured seven times and stress and anxiety were measured once and twice, respectively, as subjective stress markers. Significant changes in the level of salivary alpha-amylase were found in response to psychological stress. However, the correlations of salivary alpha-amylase with the plasma catecholamines, blood pressure, and heart rate were only partially found to be statistically significant. In conclusion, it was shown that salivary alpha-amylase was sensitive to stress throughout this study. Thus, salivary alpha-amylase may be used to measure stress uninvasively in both clinical settings and nursing research where the effects of stress might be scrutinized. Furthermore, the mechanisms of illnesses that are induced by stress could be explored. © 2010 Blackwell Publishing Asia Pty Ltd.

  10. Inhibitory Effect of Heracleum persicum and Ziziphus jujuba on Activity of Alpha-Amylase

    Directory of Open Access Journals (Sweden)

    Reza Afrisham

    2015-01-01

    Full Text Available Postprandial hyperglycemia plays an important role in the development of type 2 diabetes. Inhibition of alpha-amylase was led to a delay in breaks down of starch and glycogen and prevented a rapid rise in blood sugar. Alpha-amylase was isolated by gel filtration chromatography Sephadex G-75 from bovine pancreas. Then, total methanolic extracts of plants were prepared and IC50 values of extracts on alpha-amylase were obtained and compared with acarbose IC50. The polyphenolic content of extracts and antioxidant capacity were determined by Folin-Ciocalteu test and DPPH test, respectively. The specific activity of alpha-amylase was 48.2 U/mg. For inhibition of alpha-amylase, IC50 values of H. persicum, Z. jujuba, and acarbose were 307, 827, and 113 μg/ml, respectively. For inhibition of DPPH radical, IC50 values of extracts were 235 and 701 μg/ml. Total phenolic contents of methanol extracts were 73.8±3.2 and 44.2±1.8 μg tannic acid equivalent/mg extract. Acarbose causes gastrointestinal symptoms and liver toxicity, but H. persicum and Z. jujuba decrease these side effects and prevent gastrointestinal disorders. Due to the high polyphenolic content and antioxidant capacity of these plants and significant inhibitory effect of the plants on alpha-amylase, these plants can be proposed for treatment of diabetic patients.

  11. 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+.

  12. Salivary cortisol, salivary alpha amylase, and the dental anxiety scale.

    Science.gov (United States)

    Sadi, Hana; Finkelman, Matthew; Rosenberg, Morton

    2013-01-01

    The aim of this study was to investigate the correlation between dental anxiety, salivary cortisol, and salivary alpha amylase (sAA) levels. Furthermore, the aim was to look into individual differences such as age, race, gender, any existing pain, or traumatic dental experience and their effect on dental anxiety. This study followed a cross-sectional design and included a convenience sample of 46. Every patient was asked to complete the Dental Anxiety Scale (DAS) and a basic demographic/dental history questionnaire. A saliva sample, utilizing the method of passive drooling, was then collected in 2-mL cryovials. Samples were analyzed for salivary cortisol and sAA levels by Salimetrics. Significant associations were observed between DAS scores and presence of pain and history of traumatic dental experience. However, no significant correlations were observed between DAS, cortisol, and sAA levels. Our study reconfirms that dental anxiety is associated with presence of pain and a history of traumatic dental experience. On the other hand, our study was the first to our knowledge to test the correlation between the DAS and sAA; nevertheless, our results failed to show any significant correlation between dental anxiety, cortisol, and sAA levels.

  13. Diurnal Salivary Alpha-amylase Dynamics among Dementia Family Caregivers

    Science.gov (United States)

    Liu, Yin; Granger, Douglas A.; Kim, Kyungmin; Klein, Laura C.; Almeida, David M.; Zarit, Steven H.

    2016-01-01

    Objective The study examined diurnal regulation of salivary alpha-amylase (sAA) in association with daily stressors, adult day services (ADS) use, and other caregiving characteristics. Methods A sample of 165 family caregivers of individuals with dementia (IWD) completed an 8-day diary study. Caregivers provided 5 saliva samples across the 8 days. On some days, caregivers provided all or most of the care. On other days, their relative attended ADS for part of the day. A 3-level unconditional linear spline model was fit to describe the typical sAA diurnal rhythms. Predictors were then added to the unconditional model to test the hypotheses on ADS use and daily stressors. Results Daily ADS use did not have an effect on diurnal sAA regulation. However, controlling for daily ADS use, greater ADS use over the 8 days was associated with a more prominent rise between 30 minutes after wake-up and before lunch, and a more prominent decline between before lunch and late afternoon. Fewer ADS days were associated with a more flattened sAA diurnal rhythm. Additionally, greater daily care-related stressor exposures had a within-person association with lower sAA levels in the late afternoon. Care-related stressor exposures had significant within- and between-person associations with sAA diurnal slopes. Furthermore, daily positive experiences had a significant between-person association with sAA diurnal slopes. Conclusions Caring for a disabled family member may heighten the vulnerability to potential physiological conditions. Respite from care stressors from ADS use may have some biobehavioral benefits on sAA regulations. PMID:27786517

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

    Directory of Open Access Journals (Sweden)

    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.

  15. Long-term stability of diurnal salivary cortisol and alpha-amylase secretion patterns.

    Science.gov (United States)

    Skoluda, Nadine; La Marca, Roberto; Gollwitzer, Mario; Müller, Andreas; Limm, Heribert; Marten-Mittag, Birgitt; Gündel, Harald; Angerer, Peter; Nater, Urs M

    2017-06-01

    This study aimed to investigate long-term stability and variability of diurnal cortisol and alpha-amylase patterns. Diurnal cortisol and alpha-amylase secretion patterns were assessed on a single workday with three waves of measurement across a total time period of 24months in 189 participants. Separate hierarchical linear models were analyzed, with and without a number of potential predictor variables (age, BMI, smoking, chronic stress, stress reactivity). While low long-term stability was found in diurnal cortisol, the stability of diurnal alpha-amylase was moderate across the time period of 24months. Several predictor variables had a positive impact on diurnal cortisol and alpha-amylase secretion patterns averaged across waves. Our findings underpin the notion that long-term stability is not necessarily warranted in longitudinal studies. It is important to choose an appropriate study design when attempting to disentangle clinically and biologically relevant changes from naturally occurring variations in diurnal cortisol and alpha-amylase. Copyright © 2017 Elsevier Inc. All rights reserved.

  16. Influence of carbon source on alpha-amylase production by Aspergillus oryzae

    DEFF Research Database (Denmark)

    Carlsen, Morten; Nielsen, Jens

    2001-01-01

    on sucrose, fructose, glycerol, mannitol and acetate. During growth on acetate there was no production of alpha -amylase, whereas addition of small amounts of glucose resulted in alpha -amylase production. A possible induction by alpha -methyl-D-glucoside during growth on glucose was also investigated......, but this compound was not found to be a better inducer of alpha -amylase production than glucose. The results strongly indicate that besides acting as a repressor via the CreA protein, glucose acts as an inducer.......The influence of the carbon source on a-amylase production by Aspergillus oryzae was quantified in carbon-limited chemostat cultures. The following carbon sources were investigated: maltose, maltodextrin (different chain lengths), glucose, fructose, galactose, sucrose, glycerol, mannitol...

  17. 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)

  18. The effect of gamma irradiation on the formation of alpha-amylase isoenzymes in germinating wheat

    International Nuclear Information System (INIS)

    Machaiah, J.P.; Vakil, U.K.

    1979-01-01

    The biosynthesis of alpha-amylase during seedling growth commenced after a prolonged lag-period in wheat (cv. Vijay), irradiated at a high dose (200 krad). Also, a different requirement for exogenous gibberellins (GA) to stimulate the enzyme synthesis was noted in control and irradiated seeds. Further, the developmental patterns of three major isoenzymes of alpha-amylase (designated as α 1 , α 2 - and α 3 ) during germination were different. It was observed that α 1 -isoenzyme which appeared on the fourth day of germination of control seeds, was delayed in its development and was undetectable up to 4 days in samples irradiated with 200 krad. However, α 1 -isoenzyme appeared after 6 days or after 4 days in GA-treated samples in germinating seeds exposed to a high dose. These results suggested that two systems differing in their radiosensitivity and response to GA application were operating in germinating wheat for the synthesis of functional alpha-amylase molecules. (author)

  19. 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.

  20. Effect of pullulanase and. alpha. -amylase on hydrolysis of waxy corn starch

    Energy Technology Data Exchange (ETDEWEB)

    Sreenath, H.K. (Defence Food Research Lab., Mysore (India)); BeMiller, J. (Purdue Univ., Lafayette, IN (USA). Whistler Center for Carbohydrate Research)

    1990-12-01

    Using commercial pullulanase, {alpha}-amylase and their mixture, partial hydrolysis of waxy corn starch was characterised for optimising production of maltodextrins free of D-glucose. Compared to pullulanase or {alpha}-amylase alone, the mixture of these two (simultaneous or successive addition) on the substrate enhanced the efficiency of maltodextrin turnover with low or traces of D-glucose production in a short time. D-glucose was removed from dextrins by mambrane filtration and the yield of dextrin above DP6 was 60-65%. (orig.).

  1. 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...

  2. Exposure to inhalable dust, wheat flour and alpha-amylase allergens in industrial and traditional bakeries.

    Science.gov (United States)

    Bulat, Petar; Myny, Katrien; Braeckman, Lutgart; van Sprundel, Marc; Kusters, Edouard; Doekes, Gert; Pössel, Kerstin; Droste, Jos; Vanhoorne, Michel

    2004-01-01

    This study was designed to characterize exposure to inhalable dust, wheat flour and alpha-amylase allergens in industrial and traditional bakeries. The study included 70 bakeries from the northern part of Belgium. Based on the degree of automation and a clear division of individual job tasks, four bakeries were identified as industrial and the remaining 66 were identified as traditional ones. Personal, as well as stationary, samples of inhalable dust were collected during full shift periods, usually 5-7 h. The portable pumps aspirated 2 l/min through Teflon personal dust samplers (Millipore, pore size 1.0 microm) mounted in PAS-6 sampling heads. In the collected samples the inhalable dust, wheat flour and alpha-amylase allergens were determined. Wheat flour allergens were measured using enzyme-linked immunosorbent assay inhibition and an antiwheat IgG4 serum pool. The alpha-amylase allergens were measured using a sandwich enzyme immunoassay with affinity-purified polyclonal rabbit IgG antibodies. In total, 440 samples (300 personal and 140 stationary) were processed. The highest inhalable dust exposure was observed in traditional bakeries among bread [geometric mean (GM) 2.10 mg/m3] and bread and pastry workers (GM 1.80 mg/m3). In industrial bakeries the highest dust exposure was measured in bread-producing workers (GM 1.06 mg/m3). Similar relations were observed for wheat flour and alpha-amylase allergens. Bread baking workers in traditional bakeries had the highest exposure to both allergens (wheat flour GM 22.33 microg/m(3), alpha-amylase GM 0.61 ng/m3). The exposure to wheat flour and alpha-amylase allergens in industrial bakeries was higher in bread baking workers (wheat flour GM 6.15 microg/m3, alpha-amylase GM 0.47 ng/m3) than in bread packing workers (wheat flour GM 2.79 microg/m3, alpha-amylase GM 0.15 ng/m3). The data presented suggest that, on average, exposure in the Belgium bakeries studied-industrial as well as traditional-is lower than or similar to

  3. Ability to bind salivary alpha-amylase discriminates certain viridans group streptococcal species.

    Science.gov (United States)

    Kilian, M; Nyvad, B

    1990-01-01

    A collection of 144 viridans group streptococcal strains recently characterized as part of a taxonomic study was examined for the ability to bind salivary alpha-amylase. This property was found in most strains of Streptococcus gordonii and Streptococcus mitis and in occasional strains of Streptococcus anginosus and Streptococcus salivarius. In contrast, all strains of Streptococcus sanguis, Streptococcus oralis, Streptococcus vestibularis, and Streptococcus mutans lacked alpha-amylase-binding capacity. A rapid and easy assay described in this paper may be an important supplementary test for identification of oral streptococci. PMID:2254435

  4. Engineering of factors determining alpha-amylase and cyclodextrin glycosyltransferase specificity in the cyclodextrin glycosyltransferase from Thermoanaerobacterium thermosulfurigenes EM1

    NARCIS (Netherlands)

    Wind, RD; Buitelaar, RM; Dijkhuizen, L

    1998-01-01

    The starch-degrading enzymes alpha-amylase and cyclodextrin glycosyltransferase (CGTase) are functionally and structurally closely related, with CGTases containing two additional domains (called D and E) compared to the three domains of alpha-amylases (A, B and C). Amino acid residue 196

  5. The effect of alpha amylase enzyme on quality of sweet sorghum juice for chrystal sugar

    Science.gov (United States)

    Marwati, T.; Cahyaningrum, N.; Widodo, S.; Astiati, U. T.; Budiyanto, A.; Wahyudiono; Arif, A. B.; Richana, N.

    2018-01-01

    Sweet sorghum juice (Sorghum bicolor L. Moench) has characteristics similar to sugar cane juice and potentially used for sugar substitutes that can support food security. Nevertheless the sweet sorghum juicecontain starch which impede sorghum sugar crystallization. Therefore, research on the enzymatic process is needed to convert starch into reducing sugar. The experimental design used was the Factorial Randomized Design with the first factor was alpha amylase enzyme concentration (0, 20, 40, 60, 80, 100, 120 μL/100 mL) and second factor was incubation time (0, 30, 60, 90 minute) at temperature 100°C. The experiment was conducted on fresh sweet sorghum. The results showed that the addition of the alpha amylase enzyme increased the content of reducing sugar and decreased levels of starch. Elevating concentration of alpha amylase enzyme will increase the reducing sugar content in sweet sorghum juice. The optimum alpha amylase enzyme concentration to produce the highest total sugar was 80 μL/100 mL of sweet sorghum juice with the optimum incubation time was 90 minutes. The results of this study are expected to create a new sweetener for sugar substitution. From the economic prospective aspect, sorghum is a potential crop and can be relied upon to support the success of the food diversification program which further leads to the world food security

  6. Production of alpha-amylase from Aspergillus oryzae for several industrial applications in a single step.

    Science.gov (United States)

    Porfirif, María C; Milatich, Esteban J; Farruggia, Beatriz M; Romanini, Diana

    2016-06-01

    A one-step method as a strategy of alpha-amylase concentration and purification was developed in this work. This methodology requires the use of a very low concentration of biodegradable polyelectrolyte (Eudragit(®) E-PO) and represents a low cost, fast, easy to scale up and non-polluting technology. Besides, this methodology allows recycling the polymer after precipitation. The formation of reversible soluble/insoluble complexes between alpha-amylase and the polymer Eudragit(®) E-PO was studied, and their precipitation in selected conditions was applied with bioseparation purposes. Turbidimetric assays allowed to determine the pH range where the complexes are insoluble (4.50-7.00); pH 5.50 yielded the highest turbidity of the system. The presence of NaCl (0.05M) in the medium totally dissociates the protein-polymer complexes. When the adequate concentration of polymer was added under these conditions to a liquid culture of Aspergillus oryzae, purification factors of alpha-amylase up to 7.43 and recoveries of 88% were obtained in a simple step without previous clarification. These results demonstrate that this methodology is suitable for the concentration and production of alpha-amylase from this source and could be applied at the beginning of downstream processing. Copyright © 2016 Elsevier B.V. All rights reserved.

  7. Peer Victimization and Aggression: Moderation by Individual Differences in Salivary Cortisol and Alpha-Amylase

    Science.gov (United States)

    Rudolph, Karen D.; Troop-Gordon, Wendy; Granger, Douglas A.

    2010-01-01

    This research examined whether variations in salivary measures of the hypothalamic-pituitary-adrenal axis (cortisol) and autonomic nervous system (alpha amylase [sAA]) contribute to individual differences in the association between peer victimization and aggression. Children (N = 132; M age = 9.46 years, SD = 0.33) completed a measure of peer…

  8. changes in the composition of the pulp, alpha-amylase activity and ...

    African Journals Online (AJOL)

    DJFLEX

    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 ...

  9. A rapid lateral flow immunoassay for the detection of fungal alpha-amylase at the workplace

    NARCIS (Netherlands)

    Koets, M.; Sander, I.; Bogdanovic, J.; Doekes, G.; Amerongen, van A.

    2006-01-01

    Fungal alpha-amylase is a flour supplement which is added to improve the quality of bakery products. Various studies have shown that exposure to this enzyme is an important risk factor for the development of bakers allergy and this allergy is reported to be one of the most frequent causes of

  10. SALIVARY ALPHA-AMYLASE AS A BIOMARKER OF DENTAL FEAR AND ANXIETY IN CHILDREN

    Directory of Open Access Journals (Sweden)

    Réka GYERGYAY

    2015-03-01

    Full Text Available Dental treatment represents a stress factor for most children. The aim of the study was to analyse the variation of salivary alpha-amylase concentration in children after a video viewing on dental treatments. In this study, 7 to 10 year-old school children were evaluated (n=119. Unstimulated whole saliva was collected before and after viewing a 15 min video on dental treatments performed on children. Changes in salivary alpha-amylase levels have been assessed. Video viewing on dental procedures led to a significant increase of the alpha-amylase level in the whole sample group. This was noticeable in terms of gender as well as age groups. From the viewpoint of age and gender, girls displayed significantly higher levels of amylase in all age groups, while this could be observed only in younger boys. In conclusion, analysis of salivary alpha-amylase revealed that the sight of dental treatment represents a significant source of stress among children.

  11. 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 ...

  12. 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

  13. [Characteristics of alpha-amylase isozymes in cytologenetically different wheat cultivars].

    Science.gov (United States)

    Netsvetaev, V P; Badaeva, E D

    2014-07-01

    The isoenzyme composition of alpha-amylase is studied by polyacrylamide gel electrophoresis in Tris-glycine (pH 8.3) system in wheat cultivars with different genome composition. We show that durum wheat (Triticum durum, 2n=4x=28, BBAA) lacks the isoenzymes encoded by 6D and 7D chromosomes that are present in common wheat zymograms (Triticum aestivum, 2n=6x=42, BBAADD). A similar pattern is observed in a synthetic allohexaploid carrying the BBAA genomes of wheat and the HchHch genome of barley (Hordeum chilense). Our method of electrophoresis fails to reveal additional variants of alpha-amylase encoded by the barley genome, although C-banding analysis confirms the genomic structure BBAAHChHCh of this allopolyploid. The electrophoretic spectrum of the spring common wheat cultivar Dobrynya with the wheat-Agropyron translocation 7DL-7AiL contains all of the alpha-amylase isoenzymes typical for common wheat (2n=6x=42, BBAADD) except for the zymotype encoded by the long arm of chromosome 7D. This observation confirms the results of cytogenetic analysis that identified a 7DL-7AiL translocation in this cultivar. No additional alpha-amylase isoenzymes encoded by Agropyron chromosome have been observed. Our data indicate that analysis of wheat-alien hybrids or introgressive forms should be carried out using a complex of different methods.

  14. Aerobic Fitness Level Affects Cardiovascular and Salivary Alpha Amylase Responses to Acute Psychosocial Stress.

    Science.gov (United States)

    Wyss, Thomas; Boesch, Maria; Roos, Lilian; Tschopp, Céline; Frei, Klaus M; Annen, Hubert; La Marca, Roberto

    2016-12-01

    Good physical fitness seems to help the individual to buffer the potential harmful impact of psychosocial stress on somatic and mental health. The aim of the present study is to investigate the role of physical fitness levels on the autonomic nervous system (ANS; i.e. heart rate and salivary alpha amylase) responses to acute psychosocial stress, while controlling for established factors influencing individual stress reactions. The Trier Social Stress Test for Groups (TSST-G) was executed with 302 male recruits during their first week of Swiss Army basic training. Heart rate was measured continuously, and salivary alpha amylase was measured twice, before and after the stress intervention. In the same week, all volunteers participated in a physical fitness test and they responded to questionnaires on lifestyle factors and personal traits. A multiple linear regression analysis was conducted to determine ANS responses to acute psychosocial stress from physical fitness test performances, controlling for personal traits, behavioural factors, and socioeconomic data. Multiple linear regression revealed three variables predicting 15 % of the variance in heart rate response (area under the individual heart rate response curve during TSST-G) and four variables predicting 12 % of the variance in salivary alpha amylase response (salivary alpha amylase level immediately after the TSST-G) to acute psychosocial stress. A strong performance at the progressive endurance run (high maximal oxygen consumption) was a significant predictor of ANS response in both models: low area under the heart rate response curve during TSST-G as well as low salivary alpha amylase level after TSST-G. Further, high muscle power, non-smoking, high extraversion, and low agreeableness were predictors of a favourable ANS response in either one of the two dependent variables. Good physical fitness, especially good aerobic endurance capacity, is an important protective factor against health

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

    Directory of Open Access Journals (Sweden)

    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.

  16. 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.

  17. Bacillus subtilis PrsA is required in vivo as an extracytoplasmic chaperone for secretion of active enzymes synthesized either with or without pro-sequences

    DEFF Research Database (Denmark)

    Jabobs, Myra F; Andersen, Jens Bo; Kontinen, Vesa P.

    1993-01-01

    In prsA (protein secretion) mutants of Bacillus subtilis, decreased levels of exoproteins, including alpha-amylase and subtilisins, are found extracellularly. The effect of prsA on subtilisin secretion is elaborated here. Extracytoplasmic folding and secretion of active subtilisin is assisted......) and those without (like alpha-amylase)....

  18. Crystallization and preliminary X-ray diffraction studies of alpha-amylase from the antarctic psychrophile Alteromonas haloplanctis A23.

    Science.gov (United States)

    Aghajari, N.; Feller, G.; Gerday, C.; Haser, R.

    1996-01-01

    A cold-active alpha-amylase was purified from culture supernatants of the antarctic psychrophile Alteromonas haloplanctis A23 grown at 4 degrees C. In order to contribute to the understanding of the molecular basis of cold adaptations, crystallographic studies of this cold-adapted enzyme have been initiated because a three-dimensional structure of a mesophilic counterpart, pig pancreatic alpha-amylase, already exists. alpha-Amylase from A. haloplanctis, which shares 53% sequence identity with pig pancreatic alpha-amylase, has been crystallized and data to 1.85 A have been collected. The space group is found to be C222(1) with a = 71.40 A, b = 138.88 A, and c = 115.66 A. Until now, a three-dimensional structure of a psychrophilic enzyme is lacking. PMID:8897615

  19. Response surface methodology for the optimization of alpha amylase production by serratia marcescens SB08

    International Nuclear Information System (INIS)

    Venil, C.K.; Lakshmanaperumalsamy, P.

    2008-01-01

    In this work, central composite design combining with response surface methodology was successfully employed to optimize medium composition for the production of alpha amylase by Serratia marcescens SB08 in submerged fermentation. The process parameters that influence the enzyme production were identified using Plackett- Burman design. Among the various factors screened, inoculum concentration, pH, NaCl and CaCl/sub 2/ were found to be most significant. The optimum level of pH was 5.0, inoculum concentration 3%, NaCl 0.30 g/l and CaCl/sub 2/ 0.13 g/l. The actual enzyme yield before and after optimization was 56.43 U/ml and 87.23 U/ml, respectively. Thus, it is advisable to the microbial industry sponsors to apply such profitable bioprocess to maintain high yield for mass production of alpha amylase. (author)

  20. Digestive alpha-amylases of the flour moth Ephestia kuehniella - adaptation to alkaline environment and plant inhibitors

    Czech Academy of Sciences Publication Activity Database

    Pytelková, Jana; Hubert, J.; Lepšík, Martin; Šobotník, Jan; Šindelka, Radek; Křížková, I.; Horn, Martin; Mareš, Michael

    2009-01-01

    Roč. 276, č. 13 (2009), s. 3531-3546 ISSN 1742-464X R&D Projects: GA AV ČR IAA400550617; GA MŠk LC512; GA ČR GA301/09/1752 Institutional research plan: CEZ:AV0Z40550506; CEZ:AV0Z50520701 Keywords : alkaline adaptation * alpha - amylase * alpha - amylase inhibitor * Ephestia kuehniella * plant-insect interaction Subject RIV: CE - Biochemistry Impact factor: 3.042, year: 2009

  1. Genetically Modified alpha-Amylase Inhibitor Peas Are Not Specifically Allergenic in Mice

    OpenAIRE

    Lee, Rui-Yun; Reiner, Daniela; Dekan, Gerhard; Moore, Andrew E.; Higgins, T. J. V.; Epstein, Michelle M.

    2013-01-01

    Weevils can devastate food legumes in developing countries, but genetically modified peas (Pisum sativum), chickpeas and cowpeas expressing the gene for alpha-amylase inhibitor-1 (αAI) from the common bean (Phaseolus vulgaris) are completely protected from weevil destruction. αAI is seed-specific, accumulated at high levels and undergoes post-translational modification as it traverses the seed endomembrane system. This modification was thought to be responsible for the reported allergenicity ...

  2. Alpha-amylase inhibitory activity and phytochemical study of Zhumeria majdae Rech. f. and Wendelbo

    OpenAIRE

    Mirshafie, Behnaz; Mokhber-Dezfouli, Najmeh; Manayi, Azadeh; Saeidnia, Soodabeh; Ajani, Yousef; Gohari, Ahmad Reza

    2015-01-01

    Background: Zhumeria majdae (Lamiaceae) is an endemic species growing in the South parts of Iran especially Hormozgan province. The plant is so-called Mohrekhosh locally and widely used for medicinal purposes including stomachache and dysmenorrhea. Objective: In order to separation and identification of the main flavonoid glycosides of the plant (aerial parts including leaves, stems, flowers, and fruits were used) and evaluation of its alpha-amylase inhibitory (AAI) activity, methanolic extra...

  3. Gibberellic acid-stimulated alpha-amylase secretion and phospholipid metabolism in wheat aleurone tissue.

    OpenAIRE

    Mirbahar, R B; Laidman, D L

    1982-01-01

    1. Turnovers of [14C]glycerol-labelled phospholipids in wheat aleurone tissue have been measured by using a pulse-decay technique. The most metabolically active phospholipids were phosphatidylcholine, phosphatidylinositol, phosphatidylethanolamine and phosphatidylglycerol. 2. Gibberellic acid action on the tissue led to breakdown of phosphatidylcholine and stimulated turnover of the other phosphatides concomitant with the secretion of alpha-amylase from the tissue. After pulse-labelling of th...

  4. Identification of human saliva by antisera to alpha-amylase in human salivary glands.

    Science.gov (United States)

    Ohya, I; Iwasa, M; Komoriya, H; Bunai, Y; Sagisaka, K

    1986-11-01

    Amylase activities were detected significantly in saliva from human, macaques and rodents and slightly in the vegetable and fruit extracts. Dried stains on filter paper prepared from human and mammalian saliva, and the vegetable and fruit extracts were subjected to starch-iodine test and blue starch polymer-agar plate test. Both tests showed strong positive reactions with the macaque and rodent saliva stains as well as human, but the vegetable and fruit stains showed clear positive reactions only in the starch-iodine test. The results suggest that these tests are not specific for human saliva and that for screening test of saliva stains, blue starch polymer-agar plate test is more suitable than starch-iodine test. Rabbit antisera against alpha-amylase isolated from the human submaxillary glands were prepared. In double immunodiffusion test with the human and mammalian saliva, human and macaque saliva produced precipitation lines. Human saliva gave patterns of partial identity with saliva from Japanese monkey and crab-eating monkey and these two macaque saliva gave total identity with each other. Anti-alpha-amylase sera absorbed with Japanese monkey saliva reacted only with human saliva. This suggested that the anti-alpha-amylase sera absorbed with macaque saliva make it possible to identify human saliva stain.

  5. production of a thermostable α-amylase and its assay using bacillus ...

    African Journals Online (AJOL)

    Omoh-Sexy

    Correspondence author: .... and biochemical examination. Pure bacterial ..... of a thermostable Bacillus stearothermophilus alpha-amylase. Biotechnology. Applied. Biochemistry. 12:427– 435. Von S.F., Mayr, R. and Scherer, S. (1999). Climatic influence ...

  6. Physiological characterisation of recombinant Aspergillus nidulans strains with different creA genotypes expressing A-oryzae alpha-amylase

    DEFF Research Database (Denmark)

    Agger, Teit; Petersen, J.B.; O'Connor, S.M.

    2002-01-01

    The physiology of three strains of Aspergillus nidulans was examined-a creA deletion strain, a wild type creA genotype and a strain containing extra copies of the creA gene, all producing Aspergillus oryzae alpha-amylase. The strains were cultured in batch and continuous cultivations and the biom......The physiology of three strains of Aspergillus nidulans was examined-a creA deletion strain, a wild type creA genotype and a strain containing extra copies of the creA gene, all producing Aspergillus oryzae alpha-amylase. The strains were cultured in batch and continuous cultivations...... and the biomass formation and alpha-amylase production was characterised. Overexpression of the creA gene resulted in a lower maximum specific growth rate and a slightly higher repression of the alpha-amylase production during conditions with high glucose concentration. No expression of creA also resulted...... in a decreased maximum specific growth rate, but also in drastic changes in morphology. Furthermore, the expression of alpha-amylase was completely derepressed and creA thus seems to be the only regulatory protein responsible for glucose repression of alpha-amylase expression. The effect of different carbon...

  7. Molecular identification of four different alpha-amylase inhibitors from baru (Dipteryx alata) seeds with activity toward insect enzymes.

    Science.gov (United States)

    Bonavides, Krishna B; Pelegrini, Patrícia B; Laumann, Raúl A; Grossi-de-Sá, Maria F; Bloch, Carlos; Melo, Jorge A T; Quirino, Betania F; Noronha, Eliane F; Franco, Octávio L

    2007-07-31

    The endophytic bruchid pest Callosobruchus maculatus causes severe damage to storage cowpea seeds, leading to economical losses. For this reason the use of alpha-amylase inhibitors to interfere with the pest digestion process has been an interesting alternative to control bruchids. With this aim, alpha-amylase inhibitors from baru seeds (Dipteryx alata) were isolated by affinity chromatographic procedures, causing enhanced inhibition of C. maculatus and Anthonomus grandis alpha-amylases. To attempt further purification, this fraction was applied onto a reversed-phase HPLC column, generating four peaks with remarkable inhibition toward C. maculatus alpha-amylases. SDS-PAGE and MALDI-ToF analysis identified major proteins of approximately 5.0, 11.0, 20.0 and 55 kDa that showed alpha-amylase inhibition. Results of in vivo bioassays using artificial seeds containing 1.0% (w/w) of baru crude extract revealed 40% cowpea weevil larvae mortality. These results provide evidence that several alpha-amylase inhibitors classes, with biotechnological potential, can be isolated from a single plant species.

  8. Normative references of heart rate variability and salivary alpha-amylase in a healthy young male population

    Directory of Open Access Journals (Sweden)

    Kobayashi Hiromitsu

    2012-04-01

    Full Text Available Abstract Background This study aimed to present normative reference values of heart rate variability and salivary alpha-amylase in a healthy young male population with a particular focus on their distribution and reproducibility. Methods The short-term heart rate variability of 417 young healthy Japanese men was studied. Furthermore, salivary alpha-amylase was measured in 430 men. The average age of the subjects were 21.9 years with standard deviation of 1.6 years. Interindividual variations in heart rate variability indices and salivary alpha-amylase levels were plotted as histograms. Data are presented as the mean, median, standard deviation, coefficient of variation, skewness, kurtosis, and fifth and 95th percentiles of each physiological index. Results Mean recorded values were heart period 945.85 ms, log-transformed high frequency component 9.84 ln-ms2, log-transformed low frequency component 10.42 ln-ms2, log-transformed low frequency to high frequency ratio 0.58 ln-ratio, standard deviation of beat-to-beat interval 27.17 ms and root mean square of successive difference 37.49 ms. The mean value of raw salivary alpha-amylase was 17.48 U/mL, square root salivary alpha-amylase 3.96 sqrt[U/mL] and log-transformed salivary alpha-amylase 2.65 ln[U/mL]. Log-transformed heart rate variability indices exhibited almost symmetrical distributions; however, time-domain indices of heart rate variability (standard deviation of beat-to-beat interval and root mean square of successive difference exhibited right-skewed (positive skewness distributions. A considerable right-skewed distribution was observed for raw salivary alpha-amylase. Logarithmic transformation improved the distribution of salivary alpha-amylase, although square root transformation was insufficient. The day-to-day reproducibility of these indices was assessed using intraclass correlation coefficients. Intraclass correlation coefficients of most heart rate variability and salivary indices

  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. 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.

  11. 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.

  12. Diurnal cortisol and alpha-amylase in the daily lives of older adults with vital exhaustion.

    Science.gov (United States)

    Strahler, J; Fischer, S

    2018-03-01

    Vital exhaustion (VE) is characterised by unusual fatigue, increased irritability, and a feeling of demoralisation. It has been found a major risk factor for cardiovascular diseases, and one that is independent of subclinical or clinical manifestations of coronary heart disease or lifestyle-related risk factors. Stress-induced alterations in the hypothalamic-pituitary-adrenal (HPA) axis and sympathetic nervous system may mediate the link between VE and increased cardiovascular risk. However, no studies have yet assessed both systems simultaneously and in high-risk populations, such as older adults. A total of 72 older adults (34 women, mean age 61.7±7.3) who were free of any major physical or mental illnesses filled out the Maastricht Vital Exhaustion Questionnaire (MVEQ) and the Perceived Stress Scale (PSS). To determine cortisol and alpha-amylase, participants collected saliva samples upon awakening, +30min thereafter, and at 11am, 3pm, and 8pm. Participants with higher VE reported lower perceived stress (β=-0.515, p<0.001). Individuals reporting higher VE also exhibited more diminished cortisol concentrations across the day, although only by trend (β=-0.218, p=0.092). There was no significant association between VE and diurnal alpha-amylase activity. Moreover, women had lower diurnal cortisol (β=-0.381, p=0.004) and alpha-amylase (β=-0.329, p=0.011) when compared to men. Our findings provide initial evidence for psychosocial stress to be linked to VE in older adults, while evidence for HPA alterations remains tentative. Future research is warranted to determine whether VE related hypocortisolaemia represents a specific stage of the stress adaptation process that may put individuals at risk for incident cardiovascular diseases. Copyright © 2017 Elsevier Inc. All rights reserved.

  13. Elimination of double strand nuclease activity from S1 nuclease prepared from crude alpha amylase.

    Science.gov (United States)

    Hahn, W E; Van Ness, J

    1976-01-01

    Single strand-specific s1 nuclease prepared as previously described from crude alpha amylase by DEAE-cellulose chromatography also contains nuclease which degrades double strand nucleic acid. The double strand activity can be removed by repeating the DEAE-cellulose chromatography procedure at least two additional times. S1 nuclease prepared by this procedure does not degrade double strand sheared DNA as measured by Sephadex chromatography. Under the same conditions single strand DNA is completely degraded. Thus, S1 nuclease prepared by this procedure is suitable for use in removing single strand regions in DNA/DNA duplexes and DNA/RNA hybrids. PMID:940774

  14. 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...... of binding to target enzymes by surface plasmon resonance. Generally, the mutations had little effect on k(on), whereas the k(off) values were increased up to 67-fold. The effects on the inhibitory activity, however, were far more pronounced, and the K-i values of some mutants on the AMY2-binding side...

  15. 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.

  16. ALPHA-AMYLASE PRODUCTION FROM Aspergillus oryzae M BY SUBMERGED FERMENTATION

    Directory of Open Access Journals (Sweden)

    Suleimenova

    2016-08-01

    Full Text Available The main goal of present study was implementation of the Aspergillus oryzae M strain improved technology using earlier developed method of microorganism selection. 8 pure strains of Aspergillus fungi were screened for the production of extra cellular alpha-amylase using agar medium with starch as a substrate and incubated for 72h at 30 ºС. Zone of clearance was observed for screening of the amylolytic fungi (in mm. Aspergillus oryzae M has demonstrated the highest zone of clearance. Aspergillus oryzae M was cultivated for 42 days in submerged conditions of growth using new method of fungal cultivation. This method based on immobilizing enzymes producers on solid career in submerged conditions of growth gives the way to improve quality of filtrates, which remain clear, does not require additional filtering and easily separated from the mycelium. Moreover, it allows to prolong the process of fungal cultivation and to maintain high enzymatic activity for a long period of time. Presented method allowed increasing alpha-amylase production from 321 U/ml (before immobilization to 502 U/ml (after immobilization.

  17. [Effect of pectin substances on activity of human pancreatic alpha-amylase in vitro].

    Science.gov (United States)

    Chelpanova, T I; Vitiazev, F V; Mikhaleva, N Ia; Efimtseva, É A

    2012-06-01

    Pectin substances were extracted from food plants: sweet pepper Capsicum annuum L., carrot sowing Daucus sativus L., bulb onion Allium cepa L., white cabbage Brassica oleracea L. by two methods with acid solutions similar to gastric environment. The pectins that were extracted were characterized by Monosaccharide composition and quantitative contents of uronic acids, neutral monosaccharides, methoxy groups, protein. The inhibitory effect of all extracted pectin-protein complexes on activity of pharmaceutical drugs of human pancreatic alpha-amylase was detected. It was found that the inhibitory effect of isolated pectin substances was dependent upon the species of plant source, the manner of pectin substance extraction, the chemical composition and acting concentrations. The ability of pectin substances to suppress enzyme activity was found in a range of pectin concentrations from 0.5 up to 1.5 %. It was revealed that extracted pectin substances from bulb onion and white cabbage by acid solution with pepsin had a 2.4-3.4 times greater inhibiting effect on the human pancreatic alpha-amylase activity in comparison with pectin substances extracted by solution without pepsin from the same plant sources in high concentrations.

  18. The Effects of Red and Blue Lights on Circadian Variations in Cortisol, Alpha Amylase, and Melatonin

    Directory of Open Access Journals (Sweden)

    Mariana G. Figueiro

    2010-01-01

    Full Text Available The primary purpose of the present study was to expand our understanding of the impact of light exposures on the endocrine and autonomic systems as measured by acute cortisol, alpha amylase, and melatonin responses. We utilized exposures from narrowband long-wavelength (red and from narrow-band short-wavelength (blue lights to more precisely understand the role of the suprachiasmatic nuclei (SCN in these responses. In a within-subjects experimental design, twelve subjects periodically received one-hour corneal exposures of 40 lux from the blue or from the red lights while continuously awake for 27 hours. Results showed-that, as expected, only the blue light reduced nocturnal melatonin. In contrast, both blue and red lights affected cortisol levels and, although less clear, alpha amylase levels as well. The present data bring into question whether the nonvisual pathway mediating nocturnal melatonin suppression is the same as that mediating other responses to light exhibited by the endocrine and the autonomic nervous systems.

  19. 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.

  20. THESIS-ABSTRACT Supplementation levels of exogenous alpha-amylase in broilers diets.

    Science.gov (United States)

    Oliveira, H B; Silva, M I A; Mesquita, F R

    2017-08-17

    This study aimed to evaluate the supplementation levels of an exogenous alpha-amylase in broilers diets and compare two indicators in determining the diets energy. The experiment was divided into two parallel evaluations, being one of performance and the other of metabolism. In performance assay, 1,700 one-day-old Cobb-500 male chicks were used. The animals were distributed in 50 experimental plots and evaluated five treatments with ten replicates in a completely randomized design (CRD). The treatments were: a positive control (PC), a negative control (NC) and three alpha-amylase supplementation levels 200, 400 and 600 g/t, and the NC was formulated with 50 and 90 kcal of energy reduction in relation to the PC to the phases from 1 to 21 days and from 22 to 42 days, respectively. In the metabolism assay were used 240 animals, 150 birds for stage from 14 to 21 days and 90 birds to stage from 35 to 42 days of age and the treatments were the same as the performance assay, with six replicates per treatment in CRD. All diets of metabolism test contained the digestibility indicators Lipe ® (eucalyptus purified lignin) and chromic oxide (Cr 2 O 3 ), in concentrations of 0.05 and 1.0%, respectively. In the period from 1 to 21 days old, no significant differences were observed in weight gain (WG) (P > 0.05), however, feed intake (FI) was found higher by using 200 ppm of enzyme (P 0.05), but were observed lower FI and better FC to PC treatment (P 0.05), but there was lower FI and better FC for the PC treatment (P alpha-amylase tested was effective in increasing the metabolizable energy (ME), however, not enough to be equivalent to the ME of PC treatment, a result that was better expressed in the total collection, AMEn by Lipe ® and IDE by Lipe ® methodologies. The performance results reflect the superiority of PC treatment, pointing enzyme limitation to supply the energy deficit practiced in this work.

  1. Effects of infrared radiation, solar cooking and microwave cooking on alpha-amylase inhibitor in sorghum (Sorghum bicolor L.).

    Science.gov (United States)

    Mulimani, V H; Supriya, D

    1994-10-01

    Three domestic cooking methods were studied in alpha-amylase inhibitory activity in sorghum grains. In all the treatments, overnight soaked seeds lost amylase inhibitory activity much faster. All the three treatments reduced the inhibitory activity. Use of solar cooker for reducing amylase inhibitory activity works out very economically and efficiently. Microwave cooking eliminates amylase inhibitory activity within 5 minutes.

  2. 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

  3. Daytime Secretion of Salivary Cortisol and Alpha-Amylase in Preschool-Aged Children with Autism and Typically Developing Children

    Science.gov (United States)

    Kidd, Sharon A.; Corbett, Blythe A.; Granger, Douglas A.; Boyce, W. Thomas; Anders, Thomas F.; Tager, Ira B.

    2012-01-01

    We examined daytime salivary cortisol and salivary alpha-amylase (sAA) secretion levels and variability in preschool-aged children with autism (AUT) and typically developing children (TYP). Fifty-two subjects (26 AUT and 26 TYP) were enrolled. Salivary samples were obtained at waking, midday, and bedtime on two consecutive days at three phases…

  4. Physiological characterisation of recombinant Aspergillus nidulans strains with different creA genotypes expressing A-oryzae alpha-amylase

    DEFF Research Database (Denmark)

    Agger, Teit; Petersen, J.B.; O'Connor, S.M.

    2002-01-01

    The physiology of three strains of Aspergillus nidulans was examined-a creA deletion strain, a wild type creA genotype and a strain containing extra copies of the creA gene, all producing Aspergillus oryzae alpha-amylase. The strains were cultured in batch and continuous cultivations and the biom...

  5. Involvement of individual subsites and secondary substrate binding sites in multiple attack on amylose by barley alpha-amylase

    DEFF Research Database (Denmark)

    Kramhøft, Birte; Bak-Jensen, Kristian Sass; Mori, Haruhide

    2005-01-01

    Barley alpha-amylase 1 (AMY1) hydrolyzed amylose with a degree of multiple attack (DMA) of 1.9; that is, on average, 2.9 glycoside bonds are cleaved per productive enzyme-substrate encounter. Six AMY1 mutants, spanning the substrate binding cleft from subsites -6 to +4, and a fusion protein, AMY1...

  6. Electrospray mass spectrometry characterization of post-translational modifications of barley alpha-amylase 1 produced in yeast

    DEFF Research Database (Denmark)

    Søgaard, M; Andersen, Jens S.; Roepstorff, P

    1993-01-01

    We have used electrospray mass spectrometry (ESMS) in combination with protein chemistry and genetics to delineate post-translational modifications in yeast of barley alpha-amylase 1 (AMY1), a 45 kD enzyme crucial for production of malt, an important starting material in the manufacture of beer...

  7. 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

  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. Antidiabetic effects of natural plant extracts via inhibition of carbohydrate hydrolysis enzymes with emphasis on pancreatic alpha amylase.

    Science.gov (United States)

    Etxeberria, Usune; de la Garza, Ana Laura; Campión, Javier; Martínez, J Alfredo; Milagro, Fermín I

    2012-03-01

    The increasing prevalence of type 2 diabetes mellitus and the negative clinical outcomes observed with the commercially available anti-diabetic drugs have led to the investigation of new therapeutic approaches focused on controlling postprandrial glucose levels. The use of carbohydrate digestive enzyme inhibitors from natural resources could be a possible strategy to block dietary carbohydrate absorption with less adverse effects than synthetic drugs. This review covers the latest evidence regarding in vitro and in vivo studies in relation to pancreatic alpha-amylase inhibitors of plant origin, and presents bioactive compounds of phenolic nature that exhibit anti-amylase activity. Pancreatic alpha-amylase inhibitors from traditional plant extracts are a promising tool for diabetes treatment. Many studies have confirmed the alpha-amylase inhibitory activity of plants and their bioactive compounds in vitro, but few studies corroborate these findings in rodents and very few in humans. Thus, despite some encouraging results, more research is required for developing a valuable anti-diabetic therapy using pancreatic alpha-amylase inhibitors of plant origin.

  10. Maltose effects on barley malt diastatic power enzyme activity and thermostability at high isothermal mashing temperature: II. Alpha-amylase

    Science.gov (United States)

    Maltose, the primary product of starch degradation during mashing, has the potential as a compatible solute to affect the activity of and increase the thermostability of barley malt alpha-amylase activity at high temperatures used in mashing and temperatures above those normally used in mashing. To ...

  11. Monoclinic crystal form of Aspergillus niger alpha-amylase in complex with maltose at 1.8 A resolution

    NARCIS (Netherlands)

    Vujicic-Zagar, A.; Dijkstra, B. W.; Vujičić-Žagar, A.

    Aspergillus niger alpha-amylase catalyses the hydrolysis of alpha-1,4-glucosidic bonds in starch. It shows 100% sequence identity to the A. oryzae homologue (also called TAKA-amylase), three crystal structures of which have been published to date. Two of them belong to the orthorhombic space group

  12. Alpha-amylase Inhibition and Antioxidant Activity of Marine Green Algae and its Possible Role in Diabetes Management.

    Science.gov (United States)

    Unnikrishnan, P S; Suthindhiran, K; Jayasri, M A

    2015-10-01

    In the continuing search for safe and efficient antidiabetic drug, marine algae become important source which provide several compounds of immense therapeutic potential. Alpha-amylase, alpha-glucosidase inhibitors, and antioxidant compounds are known to manage diabetes and have received much attention recently. In the present study, four green algae (Chaetomorpha aerea, Enteromorpha intestinalis, Chlorodesmis, and Cladophora rupestris) were chosen to evaluate alpha-amylase, alpha-glucosidase inhibitory, and antioxidant activity in vitro. The phytochemical constituents of all the extracts were qualitatively determined. Antidiabetic activity was evaluated by inhibitory potential of extracts against alpha-amylase and alpha-glucosidase by spectrophotometric assays. Antioxidant activity was determined by 2,2-diphenyl-1-picrylhydrazyl, hydrogen peroxide (H2O2), and nitric oxide scavenging assay. Gas chromatography-mass spectrometry (GC-MS) analysis was carried out to determine the major compound responsible for its antidiabetic action. Among the various extracts screened, chloroform extract of C. aerea (IC50 - 408.9 μg/ml) and methanol extract of Chlorodesmis (IC50 - 147.6 μg/ml) showed effective inhibition against alpha-amylase. The extracts were also evaluated for alpha-glucosidase inhibition, and no observed activity was found. Methanol extract of C. rupestris showed notable free radical scavenging activity (IC50 - 666.3 μg/ml), followed by H2O2 (34%) and nitric oxide (49%). Further, chemical profiling by GC-MS revealed the presence of major bioactive compounds. Phenol, 2,4-bis (1,1-dimethylethyl) and z, z-6,28-heptatriactontadien-2-one were predominantly found in the methanol extract of C. rupestris and chloroform extract of C. aerea. Our results demonstrate that the selected algae exhibit notable alpha-amylase inhibition and antioxidant activity. Therefore, characterization of active compounds and its in vivo assays will be noteworthy. Four green algae were

  13. High-level expression of the native barley alpha-amylase/subtilisin inhibitor in Pichia pastoris

    DEFF Research Database (Denmark)

    Micheelsen, Pernille Ollendorff; Ostergaard, Peter Rahbek; Lange, Lene

    2008-01-01

    An expression system for high-level expression of the native Hordeum vulgare alpha-amylase/subtilisin inhibitor (BASI) has been developed in Pichia pastoris, using the methanol inducible alcohol oxidase 1 (AOX1) promoter. To optimize expression, two codon-optimized coding regions have been designed...... and expressed alongside the wild-type coding region. To ensure secretion of the native mature protein, a truncated version of the alpha mating factor secretion signal from Saccharomyces cerevisiae was used. In order to be able to compare expression levels from different clones, single insertion transformants...... generated by gene replacement of the AOX1 gene was selected by PCR screening. Following methanol induction, expression levels reached 125 mgL(-1) from the wild-type coding region while expression from the two codon-optimized variants reached 65 and 125 mgL(-1), respectively. The protein was purified...

  14. Effect of C:N ratio on alpha amylase production by Bacillus ...

    African Journals Online (AJOL)

    -) amylase exhibiting activity at a wide pH range and was relatively stable. The B. lichenformis isolate, however, produces low yields of the amylase. Our results show that the amylase production is higher in the presence of starch, with ...

  15. Immobilization of alpha-amylase produced by Bacillus circulans GRS 313

    Directory of Open Access Journals (Sweden)

    Gargi Dey

    2003-03-01

    Full Text Available A maltooligosaccharide-forming amylase from B circulans GRS 313 was immobilized by entrapment in calcium alginate beads. The immobilized activity was affected by the size of the bead and bead size of 2mm was found to be most effective for hydrolysis. Kinetics constants, Km and Vmax were estimated and were found to be affected by the bead size. The catalytic activity of the enzyme was studied in presence of various starchy residues and metal ions. HgCl2, CuSO4 and FeCl3 caused inhibition of the enzyme. The reaction conditions, pH and temperature, was optimized using response surface methodology. At the optimum pH and temperature of 4.9 and 57ºC, the apparent activity was 25.6U/g of beads, resulting in almost 2-fold increase in activity. The immobilized enzyme showed a high operational stability by retaining almost 85% of the initial activity after seventh use.Um maltooligossacarideo obtido a partir de amilase produzida por B. circulans GRS 313 foi imobilizada em alginato de sódio. A atividade enzimática foi afetada pelo tamanho da partícula. Partículas com 2mm foram as mais efetivas na hidrólise. Constantes cinética Km e Vmax foram estimadas e afetadas pelo tamanho das partículas. A atividade catalítica da enzima foi estuda na presença de diferentes tipos de amido e íons metálicos. HgCl2, CuSO4 e FeCl3 provocaram inibição na enzima. As condições de reação (temperatura e pH foram otimizadas utilizando a metodologia da superfície de resposta. Em pH ótimo de 4,9 e temperatura de 57 ºC, a atividade aparente foi de 25.6 U/g de partículas, resultando num acréscimo de mais de 2 vezes na atividade da enzima. A imobilização da enzima mostrou uma alta estabilidade operacional pela retenção de 85% de sua atividade inicial após sete ciclos de utilização.

  16. Salivary alpha amylase in on-call from home fire and emergency service personnel

    Directory of Open Access Journals (Sweden)

    Sarah J Hall

    2017-10-01

    Full Text Available The effect of working on-call from home on the sympatho-adrenal medullary system activity is currently unknown. This study had two aims, Aim 1: examine salivary alpha amylase awakening response (AAR and diurnal salivary alpha amylase (sAA profile in fire and emergency service workers who operate on-call from home following a night on-call with a call (NIGHT-CALL, a night on-call without a call (NO-CALL and an off-call night (OFF-CALL, and Aim 2: explore whether there was an anticipatory effect of working on-call from home (ON compared to when there was an off-call (OFF on the diurnal sAA profile. Participants wore activity monitors, completed sleep and work diaries and collected seven saliva samples a day for one week. AAR area under the curve with respect to ground (AUCG, AAR area under the curve with respect to increase (AUCI, AAR reactivity, diurnal sAA slope, diurnal sAA AUCG and mean 12-h sAA concentrations were calculated. Separate generalised estimating equation models were constructed for each variable of interest for each aim. For Aim 1, there were no differences between NIGHT-CALL or NO-CALL and OFF-CALL for any response variable. For Aim 2, there was no difference between any response variable of interest when ON the following night compared to when OFF the following night (n = 14. These findings suggest that there is no effect of working on-call from home on sAA, but should be interpreted with caution, as overnight data were not collected. Future research, using overnight heart rate monitoring, could help confirm these findings.

  17. [Alpha-amylase as an occupational allergen in baking industry employees].

    Science.gov (United States)

    De Zotti, R; Larese, F; Molinari, S

    1994-01-01

    In a group of 226 bakers and pastry makers and in 88 students of a training school for bakers, we evaluated skin sensitization to the common allergens, wheat and alpha amylase. Skin prick tests were positive to the enzyme in 17 exposed subjects (7.5%) and in one student with previous occupational exposure as a baker; 27 exposed subjects (11.9%) and 2 students were sensitized to wheat. Among the 42 exposed workers who complained of work-related symptoms, 12 (28.6%) cases were skin positive to amylase and 17 (42.9%) to wheat. Among the 17 workers who were positive to amylase, 16 were also sensitized to wheat and/or common allergens, 12 complained of symptoms at work but since in many cases there was a positive response to wheat, too, it is impossible to speculate on the role of each allergen in inducing symptoms. One case, with work-related rhinoconjunctivitis, had skin sensitization only to alpha amylase but no specific IgE in the serum. These findings confirm that bakers are at risk of sensitization not only to wheat allergen but also to amylase from Aspergillus oryzae. The enzyme should be included in the list of substances to be tested among bakers in whom an occupational allergy is suspected, but particular care should be taken in evaluating the cutaneous response, especially if compared to wheat wheals. Further investigations are also needed to identify the source of risk and to better define the characteristics of the enzyme and the relationship between skin reaction to amylase, sensitization to wheat and atopy.(ABSTRACT TRUNCATED AT 250 WORDS)

  18. Salivary alpha amylase in on-call from home fire and emergency service personnel.

    Science.gov (United States)

    Hall, Sarah J; Aisbett, Brad; Robertson, Samuel J; Ferguson, Sally A; Turner, Anne I

    2017-11-01

    The effect of working on-call from home on the sympatho-adrenal medullary system activity is currently unknown. This study had two aims, Aim 1: examine salivary alpha amylase awakening response (AAR) and diurnal salivary alpha amylase (sAA) profile in fire and emergency service workers who operate on-call from home following a night on-call with a call (NIGHT-CALL), a night on-call without a call (NO-CALL) and an off-call night (OFF-CALL), and Aim 2: explore whether there was an anticipatory effect of working on-call from home (ON) compared to when there was an off-call (OFF) on the diurnal sAA profile. Participants wore activity monitors, completed sleep and work diaries and collected seven saliva samples a day for one week. AAR area under the curve with respect to ground (AUC G ), AAR area under the curve with respect to increase (AUC I ), AAR reactivity, diurnal sAA slope, diurnal sAA AUC G and mean 12-h sAA concentrations were calculated. Separate generalised estimating equation models were constructed for each variable of interest for each aim. For Aim 1, there were no differences between NIGHT-CALL or NO-CALL and OFF-CALL for any response variable. For Aim 2, there was no difference between any response variable of interest when ON the following night compared to when OFF the following night ( n  = 14). These findings suggest that there is no effect of working on-call from home on sAA, but should be interpreted with caution, as overnight data were not collected. Future research, using overnight heart rate monitoring, could help confirm these findings. © 2017 The authors.

  19. A calorimetric study of solute effects on the kinetic stability of a-amylase

    DEFF Research Database (Denmark)

    Olsen, Søren Nymand; Andersen, Kim Bruno; Øgendal, Lars Holm

    2009-01-01

    In this study we evaluated the applications of isothermal titration calorimetry (ITC) to Study solute effects on the kinetics of irreversible protein denaturation. More specifically, denaturation of Bacillus Halmapalus alpha-amylase (BHA) was initiated by addition of EDTA to the calorimetric cell...

  20. PURIFICATION AND CHARACTERIZATION OF ALPHA AMYLASE FROM SEEDS OF PEARL MILLET (Pennisetum typhoides

    Directory of Open Access Journals (Sweden)

    Kerdalin Kharkrang

    2012-06-01

    Full Text Available Alpha-amylase from pearl millet (Pennisetum typhoides was purified to homogeneity through several steps involving change of pH (pH 7.0 to 6.0, lyophilization (-55 ºC, polyethylene glycol precipitation (3-15%, Sepharose 6 B column chromatography and CM-cellulose column chromatography. The 24 fold purified enzyme had specific activity of 580 units/mg protein. Native PAGE revealed a single protein band and activity staining revealed that the band corresponds to alpha amylase. SDS-PAGE also revealed a single protein band. The native molecular mass determined by gel filtration was 33 kDa. The sub-unit molecular mass determined by SDS-PAGE was 33 kDa, suggesting that enzyme is a monomeric protein. The A280/A260 ratio was 1.5, clearly indicating that the enzyme is free from bound nucleotides. When the enzyme was stored at 4ºC in absence of any additive, t1/2 was 90 days. However, when the enzyme was stored at 4ºC in the presence of acetaminophen, t1/2 increased to 121 days. The Km and Vmax values are 0.5 mg/ml and 603 µmol/min/mg, respectively. The kcat was 335 s-1. From the Arrhenius plot, the activation energy was calculated to be 6.9 kcal/mol. When the enzyme was dialyzed against EDTA (1 mM, negligible loss in activity was observed suggesting that there is no absolute requirement of a divalent cation. However, divalent cations namely, Co2+, Ca2+, Mg2+, Ni2+ at 1mM concentration, activated the enzyme activity. Mn2+ inhibited the enzyme activity at the same concentration. NH4 + showed activation of the enzyme activity, Li + and K+ showed inhibition. Na+ had no effect on the enzyme activity. NAD+ activated the enzyme activity at 1 mM concentration while ATP, ADP, NADH inhibited the enzyme activity. The latter inhibitors at 3 mM concentration showed non-competitive inhibition. The Ki values of NADH, ATP and ADP were 3.96 mM, 2.0 mM and 4.48 mM, respectively. At 5 mM concentration, glycine, fructose1,6-bisphosphate and phenylalanine brought

  1. CYCLODEXTRIN FORMATION BY THE THERMOSTABLE ALPHA-AMYLASE OF THERMOANAEROBACTERIUM THERMOSULFURIGENES EM1 AND RECLASSIFICATION OF THE ENZYME AS A CYCLODEXTRIN GLYCOSYLTRANSFERASE

    NARCIS (Netherlands)

    WIND, RD; LIEBL, W; BUITELAAR, RM; PENNINGA, D; SPREINAT, A; DIJKHUIZEN, L; BAHL, H

    Extensive characterization of the thermostable alpha-amylase of Clostridium thermosulfurogenes EM1, recently reclassified as Thermoanaerobacterium thermosulfurigenes, clearly demonstrated that the enzyme is a cyclodextrin glycosyltransferase (CGTase). Product analysis after incubation of the enzyme

  2. 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

    Recombinant barley alpha-amylase isozymes 1 and 2 were secreted by Pichia pastoris at up to 50 and 1 mg/liter, respectively, representing approximately a 50-fold increase compared to the levels of the heterologous expression by Saccharomyces cerevisiae. The cDNA clones E or pM/C encoding isozymes 1...... with a value of 45,447 calculated from the sequence, liquid chromatography/mass spectrometry of endo Lys C-generated peptides followed by tandem mass spectrometry on a nanoelectrospray ionization/mass spectrometry/mass spectrometry system identified additional recombinant isozyme 1 forms to be glycosylated...... on Thr410, N-acetylated on His1, S-glutathionylated on Cys95, or C-terminally truncated of -412RS, -411QRS, and -410LQRS. The recombinant enzymes and the alpha-amylases from barley malt closely resembled each other in enzymatic activity on insoluble Blue Starch, amylose of degree of polymerization 17...

  3. Adolescents' increasing stress response to social evaluation: pubertal effects on cortisol and alpha-amylase during public speaking.

    Science.gov (United States)

    van den Bos, Esther; de Rooij, Mark; Miers, Anne C; Bokhorst, Caroline L; Westenberg, P Michiel

    2014-01-01

    Stress responses to social evaluation are thought to increase during adolescence, which may be due to pubertal maturation. However, empirical evidence is scarce. This study is the first to investigate the relation between pubertal development and biological responses to a social-evaluative stressor longitudinally. Participants performed the Leiden Public Speaking Task twice, with a 2-year interval (N = 217; age at Time 1: 8-17 years). The results support an increase in sensitivity to social evaluation during adolescence. The overall cortisol and alpha-amylase responses increased-both between and within participants-and were more strongly related to self-reported pubertal development than to age. The cortisol response shifted from speech delivery toward anticipation. The alpha-amylase response increased in both phases. © 2013 The Authors. Child Development © 2013 Society for Research in Child Development, Inc.

  4. 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...

  5. Assessing agreement between salivary alpha amylase levels collected by passive drool and eluted filter paper in adolescents with cancer.

    Science.gov (United States)

    Ameringer, Suzanne; Munro, Cindy; Elswick, R K

    2012-07-01

    To assess the validity of filter paper (FP) against the gold standard of passive drool (PD) for collecting salivary alpha amylase as a surrogate biomarker of psychological stress in adolescents with cancer. Part of a longitudinal, descriptive study of symptoms in adolescents with cancer during chemotherapy. A pediatric hematology/oncology treatment center. 33 saliva sample pairs from nine adolescents with cancer, aged 13-18 years. Salivary alpha amylase was collected by PD and FP at four time points during a cycle of chemotherapy: days 1 (time 1) and 2 (time 2) of chemotherapy, day 7-10 (time 3), and day 1 of the next cycle (time 4). A random effects regression was used to assess the correlation between PD and FP values, and a Bland Altman analysis was conducted to assess agreement between the values. Salivary alpha amylase. The estimated correlation between PD and FP values was r = 0.91, p < 0.001. Regression results were also used to rescale FP values to the levels of the PD values because the FP values were on a different scale than the PD values. The Bland Altman analysis revealed that the agreement between the rescaled FP values and PD values was not satisfactory. Eluted FP may not be a valid method for collecting salivary alpha amylase in adolescents with cancer. Psychological stress in adolescents with cancer may be linked to negative outcomes, such as greater symptom severity and post-traumatic stress disorder. Nurses need valid, efficient, biobehavioral measures to assess psychological stress in the clinical setting.

  6. Digestive alpha-amylases of the flour moth Ephestia kuehniella - adaptation to alkaline environment and plant inhibitors

    Czech Academy of Sciences Publication Activity Database

    Pytelková, Jana; Hubert, J.; Lepšík, Martin; Šobotník, Jan; Šindelka, Radek; Křížková, I.; Horn, Martin; Mareš, Michael

    2009-01-01

    Roč. 276, Suppl. 1 (2009), s. 162-162 ISSN 1742-464X. [FEBS Congress /34/. 04.07.2009-09.07.2009, Praha] R&D Projects: GA ČR GP525/09/P600; GA AV ČR IAA400550617 Institutional research plan: CEZ:AV0Z40550506; CEZ:AV0Z50520701 Keywords : Ephestia kuehniella * alpha amylase * alkaline adaptation Subject RIV: CE - Biochemistry

  7. Alpha amylase assisted synthesis of TiO{sub 2} nanoparticles: Structural characterization and application as antibacterial agents

    Energy Technology Data Exchange (ETDEWEB)

    Ahmad, Razi; Mohsin, Mohd [Department of Biosciences, Jamia Millia Islamia, New Delhi 110025 (India); Ahmad, Tokeer [Department of Chemistry, Jamia Millia Islamia, New Delhi 110025 (India); Sardar, Meryam, E-mail: msardar@jmi.ac.in [Department of Biosciences, Jamia Millia Islamia, New Delhi 110025 (India)

    2015-02-11

    Graphical abstract: - Highlights: • Green synthesis of TiO{sub 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{sub 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{sub 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{sub 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{sub 2} nanoparticles possess potent bactericidal activity. Further, growth curve study shows effect of inhibitory concentration of TiO{sub 2} nanoparticles against S. aureus and E. coli. Confocal microscopy and TEM investigation confirm that nanoparticles were disrupting the bacterial cell wall.

  8. 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

  9. 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.

  10. Harsh discipline and behavior problems: the moderating effects of cortisol and alpha-amylase.

    Science.gov (United States)

    Chen, Frances R; Raine, Adrian; Rudo-Hutt, Anna S; Glenn, Andrea L; Soyfer, Liana; Granger, Douglas A

    2015-01-01

    Numerous studies link harsh discipline to adjustment problems in youth, yet not all individuals exposed to harsh discipline develop behavior problems. Contemporary theory suggests that this relationship could be moderated by individual differences in environmentally sensitive biological systems. This study investigated whether the interaction between hypothalamic-pituitary-adrenal (HPA) activity and autonomic nervous system (ANS) arousal moderated the link between harsh discipline and behavior problems. Three saliva samples were collected on a single day from 425 inner city youth (50% male, age 11-12 years, 80% African American) and were later assayed for cortisol (HPA) and alpha-amylase (ANS). Problem behavior was assessed by self- and parent-report using the Child Behavior Checklist. Youth also reported the level of harsh discipline that they experienced. Harsh discipline was positively associated with externalizing and internalizing problems only when there were asymmetrical profiles of HPA activity and ANS arousal. This pattern was evident for boys but not girls. Findings are discussed in relation to prevailing theories suggesting that biological susceptibility translates adversity into risk for behavior problems. Copyright © 2014 Elsevier B.V. All rights reserved.

  11. Salivary Alpha Amylase, Dental Anxiety, and Extraction Pain: A Pilot Study.

    Science.gov (United States)

    Lee, Kevin C; Bassiur, Jennifer P

    2017-01-01

    The primary intention of this study was to determine whether salivary alpha-amylase (sAA) factors or the Dental Anxiety Scale (DAS) was a better predictor of dental extraction pain. This study followed a cross-sectional design and included a convenience sample (n = 23) recruited from an outpatient oral surgery clinic. While waiting for their scheduled appointments, consenting patients completed both basic demographic/medical history questionnaires and Corah's DAS as well as submitted sublingual saliva samples. After their extractions, patients marked visual analog scales (VAS) to indicate the intensity of their intraoperative discomfort. Results of this study confirm that there is a relationship between a patient's dental anxiety and intraoperative extraction pain (r[21] = .47, P = .02). This study did not find that preoperative sAA factors (concentration and output rate) were related to either VAS extraction pain or DAS score. A strong positive relationship was observed between the concentration of sAA and the rate of sAA output (r[21] = .81, P dental anxiety has a moderate but significant correlation with intraoperative dental pain. Factors of sAA do not appear to be predictive of this experience. Therefore, simply assessing an anxious patient may be the best indication of that patient's extraction pain.

  12. Aspergillus oryzae S2 alpha-amylase production under solid state fermentation: optimization of culture conditions.

    Science.gov (United States)

    Sahnoun, Mouna; Kriaa, Mouna; Elgharbi, Fatma; Ayadi, Dorra-Zouari; Bejar, Samir; Kammoun, Radhouane

    2015-04-01

    Aspergillus oryzae S2 was assayed for alpha-amylase production under solid state fermentation (SSF). In addition to AmyA and AmyB already produced in monitored submerged culture, the strain was noted to produce new AmyB oligomeric forms, in particular a dominant tetrameric form named AmyC. The latter was purified to homogeneity through fractional acetone precipitation and size exclusion chromatography. SDS-PAGE and native PAGE analyses revealed that, purified AmyC was an approximately 172 kDa tetramer of four 42 kDa subunits. AmyC was also noted to display the same NH2-terminal amino acid sequence residues and approximately the same physico-chemical properties of AmyA and AmyB, to exhibit maximum activity at pH 5.6 and 60 °C, and to produce maltose and maltotriose as major starch hydrolysis end-products. Soyabean meal was the best substitute to yeast extract compared to fish powder waste and wheat gluten waste. AmyC production was optimized under SSF using statistical design methodology. Moisture content of 76.25%, C/N substrate ratio of 0.62, and inoculum size of 10(6.87) spores allowed maximum activity of 22118.34 U/g of dried substrate, which was 33 times higher than the one obtained before the application of the central composite design (CCD). Copyright © 2015 Elsevier B.V. All rights reserved.

  13. Salivary Alpha-Amylase Enzyme, Psychological Disorders, and Life Quality in Patients with Recurrent Aphthous Stomatitis

    Directory of Open Access Journals (Sweden)

    Juliana Andrade Cardoso

    2017-01-01

    Full Text Available Objective. The aim of this study was to evaluate stress, anxiety, and salivary alpha-amylase (SAA activity in patients with recurrent aphthous stomatitis (RAS. The impact of this disease on the life quality was also evaluated. Design. Twenty-two patients with RAS and controls, matched by sex and age, were selected. Stress and anxiety were assessed using Lipp’s Inventory of Stress Symptoms and Beck Anxiety Inventory. Life quality was assessed through the World Health Organization Quality of Life-bref (WHOQOL-BREF and the Oral Health Impact Profile-14 (OHIP-14. Saliva samples were collected in the morning and afternoon and the SAA activity was analyzed by enzymatic kinetic method. Results. No significant difference was observed between the groups regarding the SAA activity (p=0.306. Patients with RAS had higher scores of anxiety (p=0.016. The scores of WHOQOL-BREF were significantly lower in patients with RAS. The values obtained through OHIP-14 were significantly higher in these patients (p=0.002. Conclusion. RAS negatively affects the life quality. Patients with the disease have higher levels of anxiety, suggesting its association with the etiopathogenesis of RAS.

  14. Psychosocial Stress Increases Salivary Alpha-Amylase Activity Independently from Plasma Noradrenaline Levels.

    Directory of Open Access Journals (Sweden)

    Liubov Petrakova

    Full Text Available Salivary alpha-amylase activity (sAA and plasma noradrenaline (NA concentrations are often considered to be surrogate markers of sympathetic activation in response to stress. However, despite accumulating evidence for a close association between sAA and noradrenaline and other indicators of sympathetic activity, reliability and generality of this relation remains unclear. We employed the Trier Social Stress Test (TSST in order to directly compare the responses in sAA and NA to psychological stress in healthy volunteers (n = 23. The TSST significantly increased sAA and NA plasma levels with no significant differences in females and males. However, when subjects were divided according to their NA responses into low versus high responders, both groups did not significantly differ in their sAA before, during or after stress exposure. These data suggest that in response to acute psychological stress both plasma NA levels and sAA reflect sympathetic activity, however seemed to increase independently from each other.

  15. Alpha-amylase inhibitory activities of ascidians in the treatment of diabetes mellitus

    Directory of Open Access Journals (Sweden)

    Antonyraj Selva Prabhu

    2014-12-01

    Full Text Available Extracts of ten selected ascidians with a reputation of usefulness in treating diabetes were examined for alpha-amylase inhibition using an in vitro model. The extract with the highest activity was selected for further characterization. From the results ethyl acetate showed predominant amylase inhibition activity for all species and the maximum level of inhibition was recorded in Phallusia mammillata (68% at 300 µg/mL and the lowest activity was noted in Microcosmus squamiger (12% at 200 µg/mL. After preliminary results, the methanolic extract of P. mammallita were further assayed for confirmation of enzyme inhibition and the maximum results (82% were obtained at 250 µg/mL and the IC50 value of P. mammillata were evidenced at 145.0 ± 0.4 µg/g. In the present study, P. mammillata indicated the maximum α-amylase activity without toxic effects. Similarly, α-glucosidase and α-amylase inhibitor bromophenol, C6H5BrO was produced by P. mammillata.

  16. Simultaneous measurement of salivary cortisol and alpha-amylase: Application and recommendations.

    Science.gov (United States)

    Strahler, Jana; Skoluda, Nadine; Kappert, Mattes B; Nater, Urs M

    2017-12-01

    Salivary cortisol (sCort) and salivary alpha-amylase (sAA) constitute proxy measures of the two major stress response systems, i.e. the hypothalamic-pituitary-adrenal axis and the autonomic nervous system, respectively. Potentially confounding determinants of sCort and sAA may limit a reliable concurrent measurement of both biomarkers, if not adequately considered. We reviewed the most important determinants of sCort and sAA and provide recommendations for handling these potential confounders. We focused on a selection of confounders, resulting in an in-depth consideration of age, sex steroid-related factors, somatic health, acute medication, smoking, consumption of food and drinks, alcohol consumption, physical activity/fitness, and sleep. Our review further highlights the importance of the consideration of potential confounders for a reliable and valid simultaneous measurement of sCort and sAA. We further provide recommendations about which and how to handle relevant confounders in the study design or data analysis. Copyright © 2017 Elsevier Ltd. All rights reserved.

  17. The salivary alpha amylase awakening response is related to over-commitment.

    Science.gov (United States)

    Eddy, Pennie; Wertheim, Eleanor H; Hale, Matthew W; Wright, Bradley J

    2018-01-25

    We assessed the relationship between physiological and psychological measures of workplace stress as measured by the effort-reward imbalance (ERI) model, with a seldom studied sample of owner-operator dairy farmers. Dairy farmers (N = 74) self-reported ERI, over-commitment (OC), dedication, and health then provided awakening saliva samples that were used to calculate the salivary alpha amylase awakening response (sAA-AR), cortisol awakening response (CAR), and salivary immunoglobulin A (sIgA) level. ERI, OC, and dedication levels were not related with sIgA or the CAR, but more over-committed farmers had a less pronounced sAA-AR. OC was more associated than ERI with the physiological indicators of stress, potentially due to the owner-operator sample used in this investigation. The suitability of sAA as a viable physiological measure of autonomic nervous system activity has been debated, but our findings promote its inclusion in future occupational stress research.

  18. 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.

  19. 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.

  20. Rapid detection of fungal alpha-amylase in the work environment with a lateral flow immunoassay.

    Science.gov (United States)

    Bogdanovic, Jelena; Koets, Marjo; Sander, Ingrid; Wouters, Inge; Meijster, Tim; Heederik, Dick; van Amerongen, Aart; Doekes, Gert

    2006-11-01

    Occupational allergen exposure assessment usually requires airborne dust sampling at the worksite followed by dust extraction and enzyme immunoassay (EIA) analysis at the laboratory. Use of semiquantitative lateral flow immunoassays (LFIAs) may allow a more rapid detection procedure with direct on-site demonstration of a bioallergen exposure hazard. In a field study, we evaluated a recently developed LFIA for fungal alpha-amylase, an important bakery allergen. Airborne and surface dust (wipe) samples and samples from flours and baking additives used at the workplace were collected in 5 industrial bakeries and tested in the LFIA for fungal amylase. For comparison, amylase was measured in sample eluates with the reference EIA method. Sensitivity of the LFIA was 1 to 10 ng/mL, and of EIA, approximately 25 pg/mL. In LFIA, most flour samples, 84% of wipe samples, 26% of personal airborne dust, and none of the 26 ambient air dust samples produced a visible reaction. Wipe samples from dough-making areas and flour samples gave the strongest reactions. All extracts with >5 ng allergen per milliliter showed a positive LFIA reaction. The LFIA for fungal amylase is an easy and rapid method to demonstrate the allergen directly at the worksite in less than 10 to 20 minutes. Similar LFIA methods may be used for other occupational allergens in other work environments. Lateral flow immunoassays for occupational allergens may be of great value in occupational hygiene surveys to demonstrate directly to workers and supervisors the hazards of work-related bioallergen exposure.

  1. Salivary alpha amylase and salivary cortisol response to fluid consumption in exercising athletes

    Directory of Open Access Journals (Sweden)

    TP Backes

    2015-11-01

    Full Text Available The objective of the study was to examine salivary biomarker response to fluid consumption in exercising athletes. Exercise induces stress on the body and salivary alpha amylase (sAA and salivary cortisol are useful biomarkers for activity in the sympathoadrenal medullary system and the hypothalamic pituitary adrenal axis which are involved in the stress response. Fifteen college students were given 150 ml and 500 ml of water on different days and blinded to fluid condition. The exercise protocol was identical for both fluid conditions using absolute exercise intensities ranging from moderate to high. Saliva was collected prior to exercise, post moderate and post high intensities and analyzed by Salimetrics assays. Exercise was significant for sAA with values different between pre-exercise (85 ± 10 U • ml-1 and high intensity (284 ± 30 U • ml-1 as well as between moderate intensity (204 ± 32 U • ml-1 and high intensity. There was no difference in sAA values between fluid conditions at either intensity. Exercise intensity and fluid condition were each significant for cortisol. Cortisol values were different between pre-exercise (0.30 ± 0.03 ug • dL -1 and high intensity (0.45 ± 0.05 ug • dL -1 as well as between moderate intensity (0.33 ± 0.04 ug • dL -1 and high intensity. Moderate exercise intensity cortisol was lower in the 500 ml condition (0.33 ± 0.03 ug • dL -1 compared with the 150 ml condition (0.38 ± 0.03 ug • dL -1 . This altered physiological response due to fluid consumption could influence sport performance and should be considered. In addition, future sport and exercise studies should control for fluid consumption.

  2. ALPHA AMYLASE AS A SALIVARY BIOMARKER OF ACUTE STRESS OF VENEPUNCTURE FROM PERIODIC MEDICAL EXAMINATIONS

    Directory of Open Access Journals (Sweden)

    David eKoh

    2014-08-01

    Full Text Available Periodic occupational health examinations often require venepuncture. Acute psychological and physical stressors during such a procedure result in sympathetic stimulation and increased salivary protein secretion, including salivary alpha-amylase (SAA. We studied SAA response to venepuncture during such an examination. Fifty eight healthy males undergoing periodic medical examination reported perceived stress level (PSL scores (on a 5-point scale and provided passive drool saliva samples at 15-min (T1 and 1-min before (T2; and 1-min (T3 and 15-min after venepuncture (T4. A subset of 33 participants available for repeat examination on a control day when there was no venepuncture provided saliva samples at the corresponding times for comparison. Saliva SAA activity levels were analysed using a salivary α-amylase assay kit (Salimetrics LLC, USAAmong 58 participants, mean SAA increased from T1 (89.95 U/L to T2 (109.5 U/L and T3 (116.9 U/L. SAA remained elevated 15 minutes after venepuncture (121.0 U/L. A positive trend in the difference of SAA between T3 and T1 was noted among subjects with increasing mean PSL scores. T3-T1 values were 0.6 (among those with PSL ≤ 1, n=24, 11.3 (among those with PSL between 1 and 1.5, n= 18 and 78.9 (among those with PSL >1.5, n=16. SAA increment over 4 time points was significantly higher on the venepuncture compared to the control day (p=0.021. SAA increases in response to the acute stress of venepuncture during a periodic medical examination, and remains elevated 15 minutes after the procedure. In comparison, such fluctuations in SAA were not seen on a control day. During venepuncture, increase in SAA from baseline is higher among those who reported greater self-perceived stress during the procedure.

  3. Reflection on design and testing of pancreatic alpha-amylase inhibitors: an in silico comparison between rat and rabbit enzyme models

    Directory of Open Access Journals (Sweden)

    Khalil-Moghaddam Shiva

    2012-11-01

    Full Text Available Abstract Background Inhibitors of pancreatic alpha-amylase are potential drugs to treat diabetes and obesity. In order to find compounds that would be effective amylase inhibitors, in vitro and in vivo models are usually used. The accuracy of models is limited, but these tools are nonetheless valuable. In vitro models could be used in large screenings involving thousands of chemicals that are tested to find potential lead compounds. In vivo models are still used as preliminary mean of testing compounds behavior in the whole organism. In the case of alpha-amylase inhibitors, both rats and rabbits could be chosen as in vivo models. The question was which animal could present more accuracy with regard to its pancreatic alpha-amylase. Results As there is no crystal structure of these enzymes, a molecular modeling study was done in order to compare the rabbit and rat enzymes with the human one. The overall result is that rabbit enzyme could probably be a better choice in this regard, but in the case of large ligands, which could make putative interactions with the −4 subsite of pancreatic alpha-amylase, interpretation of results should be made cautiously. Conclusion Molecular modeling tools could be used to choose the most suitable model enzyme that would help to identify new enzyme inhibitors. In the case of alpha-amylase, three-dimensional structures of animal enzymes show differences with the human one which should be taken into account when testing potential new drugs.

  4. Production of bacterial amylase by Bacillus species isolated from ...

    African Journals Online (AJOL)

    Optimum pH activity was obtained at 4.0 with a concentration of 0.376 mg/ml. Bacillus licheniformis has the greatest potential for producing amylase than the other isolates and rice husk can be exploited for amylase production. The B. licheniformis strain produced thermostable alpha-amylase with characteristics suitable for ...

  5. Characterization of alpha-amylase and pullulanase activities of Clostridium thermohydrosulfuricum. Evidence for a novel thermostable amylase.

    OpenAIRE

    Melasniemi, H

    1987-01-01

    Thermostable extracellular alpha-amylase and pullulanase activities of Clostridium thermohydrosulfuricum E 101-69 were characterized in a crude enzyme preparation. The activities responded similarly to temperature and pH, with optima at 85-90 degrees C and pH 5.6. The activities were stable at 65 degrees C, but were inactivated gradually in an identical manner at higher temperatures in the absence of Ca2+ and substrate. Ca2+ stabilized both activities similarly at high temperatures. Ca2+ also...

  6. Cortisol and salivary alpha-amylase trajectories following a group social-evaluative stressor with adolescents.

    Science.gov (United States)

    Katz, Deirdre A; Peckins, Melissa K

    2017-12-01

    Intraindividual variability in stress responsivity and the interrelationship of multiple neuroendocrine systems make a multisystem analytic approach to examining the human stress response challenging. The present study makes use of an efficient social-evaluative stress paradigm - the Group Public Speaking Task for Adolescents (GPST-A) - to examine the hypothalamic-pituitary-adrenocortical (HPA)-axis and Autonomic Nervous System (ANS) reactivity profiles of 54 adolescents with salivary cortisol and salivary alpha-amylase (sAA). First, we account for individuals' time latency of hormone concentrations between individuals. Second, we use a two-piece multilevel growth curve model with landmark registration to examine the reactivity and recovery periods of the stress response separately. This analytic approach increases the models' sensitivity to detecting trajectory differences in the reactivity and recovery phases of the stress response and allows for interindividual variation in the timing of participants' peak response following a social-evaluative stressor. The GPST-A evoked typical cortisol and sAA responses in both males and females. Males' cortisol concentrations were significantly higher than females' during each phase of the response. We found no gender difference in the sAA response. However, the rate of increase in sAA as well as overall sAA secretion across the study were associated with steeper rates of cortisol reactivity and recovery. This study demonstrates a way to model the response trajectories of salivary biomarkers of the HPA-axis and ANS when taking a multisystem approach to neuroendocrine research that enables researchers to make conclusions about the reactivity and recovery phases of the HPA-axis and ANS responses. As the study of the human stress response progresses toward a multisystem analytic approach, it is critical that individual variability in peak latency be taken into consideration and that accurate modeling techniques capture

  7. Amelioration in secretion of hyperthermostable and Ca2+ -independent alpha-amylase of Geobacillus thermoleovorans by some polyamines and their biosynthesis inhibitor methylglyoxal-bis-guanylhydrazone.

    Science.gov (United States)

    Uma Maheswar Rao, J L; Satyanarayana, T

    2004-01-01

    Effect of polyamines and their biosynthesis inhibitors on the production of hyperthermostable and Ca2+ -independent alpha-amylase by Geobacillus thermoleovorans MTCC 4220. The alpha-amylase was produced in starch-yeast extract-tryptone (SYT) broth with different polyamines (PA) and polyamine biosynthesis inhibitors, methylglyoxal-bis-guanylhydrazone (MGBG) and cyclohexylammonium sulphate (CHA) at 70 degrees C. The bacterial pellets were obtained after growing G. thermoleovorans at different temperatures, and used in determining total PA. The cell-free culture filtrates were used in alpha-amylase assays. During growth, total polyamines in biomass increased till 2 h, and thereafter, decreased gradually. The total polyamine content was very high in the biomass cultivated at 55 degrees C when compared with that of higher temperatures. Enzyme titre enhanced up to 70 degrees C, and thereafter declined. Extracellular enzyme and protein levels declined in the presence of exogenously added PA. The intracellular enzyme titres, however, were higher in putrescine (put) and spermidine (spd) than in spermine (spm). Polyamine biosynthesis inhibitor, MGBG enhanced secretion of alpha-amylase in a laboratory fermentor as well as shake flasks, although CHA did not affect it. The intracellular accumulation of put in the presence of MGBG appeared to enhance synthesis and secretion of alpha-amylase. Extracellular enzyme and protein levels were low in the presence of exogenously added PA, but their intracellular levels, however, were higher in put and spd than in spm. A substantial increase in the synthesis and secretion of alpha-amylase was attained in G. thermoleovorans in the presence of polyamine biosynthesis inhibitor MGBG.

  8. Comparing Dental Stress in New Child Patients and Returning Patients Using Salivary Cortisol, Immunoglobulin-A and Alpha- Amylase.

    Science.gov (United States)

    Alaki, Sumer M; Safi, Ayman; Ouda, Soliman; Nadhreen, Alaa

    this study was aimed at comparing dental stress in children having their first dental visit to those returning for dental treatment using salivary biomarkers of stress including salivary cortisol (s-cortisol), Immunoglobulin-A (s-IgA) and alpha-amylase (s-α-amylase). Additionally, the study was aimed at monitoring the change in stress in new patients as they progressed from the waiting to the clinical areas. salivary samples were collected from 40 children who had not been to a dentist before and similar samples were collected from 40 children who were returning for completion of dental treatment. Salivary cortisol, s-IgA and s-α-amylase concentrations were obtained by Enzyme-linked Immunosorbent Assay (ELISA). salivary cortisol levels were higher for new patients at the waiting area compared to that at the dental chair (p=0.05). Salivary alpha-amylase significantly increased in new patients while being seated in the dental chair. Returning patients had higher s-α-amylase (p=0.001) and s-IgA (p=0.016) compared to new patients. Returning patients had the lowest level of s-cortisol when providers were faculty pediatric dentists than with students and interns (p=0.035). children coming in for their first dental visit may experience dental stress at the waiting area before being seated for dental examination. Returning children may experience higher levels of stress compared to new child patients possibly due to previous dental exposure.

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

    Directory of Open Access Journals (Sweden)

    Pytelková Jana

    2012-01-01

    Full Text Available Abstract Background Enzymatic allergens of storage mites that contaminate stored food products are poorly characterized. We describe biochemical and immunological properties of the native alpha-amylase allergen Aca s 4 from Acarus siro, a medically important storage mite. Results A. siro produced a high level of alpha-amylase activity attributed to Aca s 4. This enzyme was purified and identified by protein sequencing and LC-MS/MS analysis. Aca s 4 showed a distinct inhibition pattern and an unusual alpha-amylolytic activity with low sensitivity to activation by chloride ions. Homology modeling of Aca s 4 revealed a structural change in the chloride-binding site that may account for this activation pattern. Aca s 4 was recognized by IgE from house dust mite-sensitive patients, and potential epitopes for cross-reactivity with house dust mite group 4 allergens were found. Conclusions We present the first protein-level characterization of a group 4 allergen from storage mites. Due to its high production and IgE reactivity, Aca s 4 is potentially relevant to allergic hypersensitivity.

  10. Alpha amylase from a fungal culture grown on oil cakes and its properties

    Directory of Open Access Journals (Sweden)

    Sumitra Ramachandran

    2004-06-01

    Full Text Available Solid-state fermentation was carried out for the production of alpha-amylase using Aspergillus oryzae. Different oil cakes such as coconut oil cake (COC sesame oil cake (SOC, groundnut oil cake (GOC, palm kernel cake (PKC and olive oil cake (OOC were screened to be used as substrate for the enzyme production and also compared with wheat bran (WB. GOC was found to be the best producer of the enzyme among these. Combination of WB and GOC (1:1 resulted higher enzyme titres than the individual substrates. Maximum amount of enzyme (9196 U/gds was obtained when SSF was carried out using WB + GOC, having initial moisture of 64% and supplemented with lactose and ammonium nitrate (1% each at 30ºC for 72h using 2 mL spore suspension (6x10(7spores/ml. Partial purification of the enzyme using ammonium sulphate fractionation resulted in 2.4-fold increase in the activity. The enzyme showed molecular weight of 68 KDa by SDS-PAGE. Except Mn, all other metal ions such as Ca, K, Na, Mg were found to be inhibitory for the enzyme activity. The enzyme was optimally active at 50(0C and pH 5.0.Fermentação no Estado Sólido foi empregada na produção de alfa-amilase usando Aspergillus niger. Diferentes tipos de torta foram utilizadas, como torta de óleo de coco (COC, torta de de óleo de amendoim (GOC torta de óleo de sesamo (SOC, torta de palma (PKC e torta de óleo de oliva (OOC foram selecionadas para serem usadas como substratos para produção de enzima e comparadas com o farelo de trigo (WB, GOC foi escolhido por ser o que produziu maiores concentrações de enzima. A combinação WB e GOC (1:1 resultou em maiores títulos da enzima quando em comparação com os substratos individuais. A máxima concentração de enzima (9196 U/ gms foi obtida quando a FES foi conduzida utilizando WB + GOC, com umidade de 64% e suplementada com lactose e nitrato de amônia (1% cada a 300C por 72 horas utilizando 2 mL de uma suspensão de esporo (6x107sporos/ml. A purifica

  11. 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

  12. 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…

  13. Salivary Alpha Amylase and Cortisol Levels in Children with Global Developmental Delay and Their Relation with the Expectation of Dental Care and Behavior during the Intervention

    Science.gov (United States)

    dos Santos, Marcio Jose Possari; Bernabe, Daniel Galera; Nakamune, Ana Claudia de Melo Stevanato; Perri, Silvia Helena Venturoli; de Aguiar, Sandra Maria Herondina Coelho Avila; de Oliveira, Sandra Helena Penha

    2012-01-01

    The purpose of this study was to analyze the alpha-amylase (sAA) and cortisol levels in children with Global developmental delay (GDD) before and after dental treatment and its association with the children's behavior during treatment. The morning salivary cortisol levels and activity of sAA of 33 children with GDD were evaluated before and after…

  14. 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...

  15. Messenger RNAs from the Scutellum and Aleurone of Germinating Barley Encode (1-->3,1-->4)-beta-d-Glucanase, alpha-Amylase and Carboxypeptidase

    DEFF Research Database (Denmark)

    Mundy, John; Brandt, Anders; Fincher, Geoffrey B

    1985-01-01

    Polyclonal antibodies raised against barley (1-->3,1-->4)-beta-d-glucanase, alpha-amylase and carboxypeptidase were used to detect precursor polypeptides of these hydrolytic enzymes among the in vitro translation products of mRNA isolated from the scutellum and aleurone of germinating barley....... In the scutellum, mRNA encoding carboxypeptidase appeared to be relatively more abundant than that encoding alpha-amylase or (1-->3,1-->4)-beta-d-glucanase, while in the aleurone alpha-amylase and (1-->3,1-->4)-beta-d-glucanase mRNAs predominated. The apparent molecular weights of the precursors for (1......-->3,1-->4)-beta-d-glucanase, alpha-amylase, and carboxypeptidase were 33,000, 44,000, and 35,000, respectively. In each case these are slightly higher (1,500-5,000) than molecular weights of the mature enzymes. Molecular weights of precursors immunoprecipitated from aleurone and scutellum mRNA translation...

  16. Biased mutagenesis in the N-terminal region by degenerate oligonucleotide gene shuffling enhances secretory expression of barley alpha-amylase 2 in yeast

    DEFF Research Database (Denmark)

    Fukuda, Kenji; Jensen, Malene Hillerup; Aghajari, Nushin

    2005-01-01

    Recombinant barley alpha-amylase 1 (rAMY1) and 2 (rAMY2), despite 80% sequence identity, are produced in very different amounts of 1.1 and alpha loop 2 that interacts with domain B (beta-->alpha loop 3) protruding from the catalytic (beta/alpha)(8)-barrel. Most remarkably Pichia pastoris strain GS...

  17. Expositie aan stof, tarwe-allergenen en schimmel alpha-amylase : stand der techniek in ambachtelijke bakkerijen, industriële bakkerijen, meelmaalderijen en bij bakkerijgrondstoffenleveranciers

    NARCIS (Netherlands)

    Pater, N. de; Doekes, G.; Miedema, E.; Goede, H.; Hemmen,J. van; Heederik, D.

    2003-01-01

    Beroepsmatige blootstelling aan meelstof kan leiden tot klachten aan de luchtwegen zoals astma en rhinitis. Deze klachten worden toegeschreven aan de in het stof aanwezige tarwe-allergenen en schimmel alpha-amylase. Het doel van het onderzoek is op basis van de resultaten een taakstelling te

  18. Two Secondary Carbohydrate Binding Sites on the Surface of Barley alpha-Amylase 1 Have Distinct Functions and Display Synergy in Hydrolysis of Starch Granules

    DEFF Research Database (Denmark)

    Nielsen, Morten Munch; Bozonnet, Sophie; Seo, Eun-Seong

    2009-01-01

    Some polysaccharide processing enzymes possess secondary carbohydrate binding sites situated on the surface far from the active site. In barley alpha-amylase 1 (AMY1), two such sites, SBS1 and SBS2, are found on the catalytic (beta/alpha)8-barrel and the noncatalytic C-terminal domain, respective...

  19. Effects of sorghum (Sorghum bicolor (L.) Moench) tannins on alpha-amylase activity and in vitro digestibility of starch in raw and processed flours

    Science.gov (United States)

    The effect of condensed tannins (CT) on in vitro starch digestibility in cooked, wholegrain sorghum flours and on corn starch was investigated. CT extracts were also tested for their inhibitory effect on alpha-amylases. Rapidly digestible starch, slowly digestible starch, and resistant starch were n...

  20. 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.

  1. Daily Social Interactions and the Biological Stress Response: Are There Age Differences in Links Between Social Interactions and Alpha-Amylase?

    Science.gov (United States)

    Birditt, Kira S; Tighe, Lauren A; Nevitt, Michael R; Zarit, Steven H

    2017-12-12

    According to the strength and vulnerability integration (SAVI) model, older people are better able to avoid negative social interactions than younger people, but when they do experience negative interactions, they are equally or more emotionally and physiologically reactive than younger people. Less is known about the links between daily negative and positive social encounters and the sympathetic adrenal medullary system (a key stress pathway) and whether there are age differences in these links. This study considers whether negative and positive social interactions are associated with diurnal alpha-amylase (a measure of the sympathetic adrenal medullary system) and whether there are differences in these links by age. Participants were from the Daily Health, Stress, and Relationship Study, which includes a random sample of 89 individuals (aged 40-95) who completed 14 days of daily diary interviews and provided saliva samples four times a day (wake, 30 min after wake, lunch, and bedtime) for four of those days that were assayed for alpha-amylase. Days in which people reported more negative interactions were associated with flatter morning declines in alpha-amylase, indicating greater stress. Links between positive interactions and diurnal alpha-amylase varied by age group. Findings are consistent with the SAVI model indicating that older adults respond differently to social stimuli than younger people. © The Author(s) 2017. Published by Oxford University Press on behalf of The Gerontological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  2. Purification and characterization of novel raw-starch-digesting and cold-adapted alpha-amylases from Eisenia foetida.

    Science.gov (United States)

    Ueda, Mitsuhiro; Asano, Tomohiko; Nakazawa, Masami; Miyatake, Kazutaka; Inouye, Kuniyo

    2008-05-01

    Novel raw-starch-digesting and cold-adapted alpha-amylases (Amy I and Amy II) from the earthworm Eisenia foetida were purified to electrophoretically homogeneous states. The molecular weights of both purified enzymes were estimated to be 60,000 by SDS-PAGE. The enzymes were most active at pH 5.5 and 50 degrees C and stable at pH 7.0-9.0 and 50-60 degrees C. Both Amy I and II exhibited activities at 10 degrees C. The enzymes were inhibited by metal ions Cu(2+), Fe(2+), and Hg(2+), and hydrolyzed raw starch into glucose, maltose and maltotriose as end products.

  3. Major water-soluble polyphenols, proanthocyanidins, in leaves of persimmon (Diospyros kaki) and their alpha-amylase inhibitory activity.

    Science.gov (United States)

    Kawakami, Kayoko; Aketa, Saiko; Nakanami, Mitsuhiro; Iizuka, Shinzo; Hirayama, Masao

    2010-01-01

    The amounts and compositions of polyphenol in persimmon leaves and persimmon leaf tea were investigated. The predominant polyphenols in fresh leaves were water-soluble, and the contents reached a maximum (2.40% w/w) in June, and then gradually decreased. Separation of them followed by thiolytic degradation revealed that the major components were unique proanthocyanidin oligomers consisting of four heterogeneous extension units, including epigallocatechin-3-O-gallate. Persimmon leaf tea also contained similar proanthocyanidins with similar compositional units. Oral administration of starch with polyphenol concentrate of persimmon leaf tea resulted in a significant and dose-dependent decrease in the blood glucose level in Wistar rats. This effect is considered to be due to inhibition of pancreas alpha-amylase. These results indicate that persimmon leaf tea containing peculiar proanthocyanidins has a significant role in suppressing blood glucose elevation after starch intake, and that the best harvest time is June.

  4. Alpha-amylase production by Streptomyces erumpens MTCC 7317 in solid state fermentation using response surface methodology (RSM).

    Science.gov (United States)

    Kar, Shaktimay; Ray, Ramesh C; Mohapatra, Uma B

    2008-01-01

    Production of alpha-amylase under solid state fermentation by Streptomyces erumpens MTCC 7317 has been investigated using different agro-industrial residues, i.e. cassava bagasse, sugarcane bagasse and wheat bran; wheat bran was found to be the best substrate. Among different nitrogen source supplemented to wheat bran, beef extract or peptone (1%) showed maximum enzyme production. Response surface methodology was used to evaluate the effect of main process parameters as incubation period (48 h), moisture holding capacity (70%), pH (7.0) and temperature (50 degrees C) on enzyme production by applying a full factorial central composite design. The maximum hydrolysis of soluble starch (90%) and cassava starch (75%) was obtained with the application of 4 ml (approximately 12096 U) of S. erumpens crude enzyme after 5 h of incubation.

  5. PRELIMINARY DATA REGARDING THE KINETIC PROPERTIES OF AN ALPHA-AMYLASE FROM ROBINIA PSEUDACACIA L. GERMINATED SEEDS

    Directory of Open Access Journals (Sweden)

    Vlad Artenie

    2005-08-01

    Full Text Available We have accomplished a partial purification of a alpha amylase from germinated seeds of Robinia pseudacacia L. by affinity precipitation. The key element is the sodium alginate, a polymer that proved affinity for this enzyme, and also has the propriety to reversibly precipitate with Ca2+. The enzyme binds to the alginate and the complex is precipitated with Ca2+. The amylase activity is recovered by dissolving the precipitate in 1M maltose and precipitating the alginate alone by addition of Ca2+. The  enzyme has a molecular weight estimated between 50 and 65 kDa, an optimum pH between 5 and 6; it is inhibited by ammonium sulfate and activated by CaCl2.

  6. Salivary alpha-amylase during pregnancy: diurnal course and associations with obstetric history, maternal demographics, and mood.

    Science.gov (United States)

    Giesbrecht, Gerald F; Granger, Douglas A; Campbell, Tavis; Kaplan, Bonnie

    2013-03-01

    Diurnal patterns of salivary alpha amylase (sAA) in pregnant women have not previously been described. The current study employed ecological momentary assessment to examine the association between the diurnal sAA, obstetric history, maternal demographics, and mood during pregnancy. Saliva was self-collected by 83 pregnant women (89% White, age 25.3-43.0 years; mean gestational age 21.9 weeks, range 6-37 weeks; gravida 1-6) at home over three days. Results indicated that current pregnancy (gestational age and fetal sex) and maternal demographics were not related to diurnal sAA. In contrast, a history of previous miscarriage (Parameter = -.17; SE = .05; p history, trait anxiety (Parameter = .16; SE = .04; p pregnancy is sensitive to emotional arousal. Evaluating diurnal patterns of sAA holds promise for advancing understanding of how emotional arousal during pregnancy may affect fetal development. Copyright © 2012 Wiley Periodicals, Inc.

  7. 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 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.

  8. 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.

  9. The impact of single nucleotide polymorphism in monomeric alpha-amylase inhibitor genes from wild emmer wheat, primarily from Israel and Golan

    Directory of Open Access Journals (Sweden)

    Yan Ze-Hong

    2010-06-01

    Full Text Available Abstract Background Various enzyme inhibitors act on key insect gut digestive hydrolases, including alpha-amylases and proteinases. Alpha-amylase inhibitors have been widely investigated for their possible use in strengthening a plant's defense against insects that are highly dependent on starch as an energy source. We attempted to unravel the diversity of monomeric alpha-amylase inhibitor genes of Israeli and Golan Heights' wild emmer wheat with different ecological factors (e.g., geography, water, and temperature. Population methods that analyze the nature and frequency of allele diversity within a species and the codon analysis method (comparing patterns of synonymous and non-synonymous changes in protein coding sequences were used to detect natural selection. Results Three hundred and forty-eight sequences encoding monomeric alpha-amylase inhibitors (WMAI were obtained from 14 populations of wild emmer wheat. The frequency of SNPs in WMAI genes was 1 out of 16.3 bases, where 28 SNPs were detected in the coding sequence. The results of purifying and the positive selection hypothesis (p Conclusions Great diversity at the WMAI locus, both between and within populations, was detected in the populations of wild emmer wheat. It was revealed that WMAI were naturally selected for across populations by a ratio of dN/dS as expected. Ecological factors, singly or in combination, explained a significant proportion of the variations in the SNPs. A sharp genetic divergence over very short geographic distances compared to a small genetic divergence between large geographic distances also suggested that the SNPs were subjected to natural selection, and ecological factors had an important evolutionary role in polymorphisms at this locus. According to population and codon analysis, these results suggested that monomeric alpha-amylase inhibitors are adaptively selected under different environmental conditions.

  10. Tyrosine 105 and threonine 212 at outermost substrate binding subsites -6 and +4 control substrate specificity, oligosaccharide cleavage patterns, and multiple binding modes of barley alpha-amylase 1

    DEFF Research Database (Denmark)

    Bak-Jensen, K.S.; André, G.; Gottschalk, T.E.

    2004-01-01

    The role in activity of outer regions in the substrate binding cleft in alpha-amylases is illustrated by mutational analysis of Tyr(105) and Thr(212) localized at subsites - 6 and +4 ( substrate cleavage occurs between subsites -1 and +1) in barley alpha-amylase 1 (AMY1). Tyr(105) is conserved...... in plant alpha-amylases whereas Thr(212) varies in these and related enzymes. Compared with wild-type AMY1, the subsite -6 mutant Y105A has 140, 15, and

  11. The spectrum of low molecular weight alpha-amylase/protease inhibitor genes expressed in the US bread wheat cultivar Butte 86

    Directory of Open Access Journals (Sweden)

    Vensel William H

    2011-07-01

    Full Text Available Abstract Background Wheat grains accumulate a variety of low molecular weight proteins that are inhibitors of alpha-amylases and proteases and play an important protective role in the grain. These proteins have more balanced amino acid compositions than the major wheat gluten proteins and contribute important reserves for both seedling growth and human nutrition. The alpha-amylase/protease inhibitors also are of interest because they cause IgE-mediated occupational and food allergies and thereby impact human health. Results The complement of genes encoding alpha-amylase/protease inhibitors expressed in the US bread wheat Butte 86 was characterized by analysis of expressed sequence tags (ESTs. Coding sequences for 19 distinct proteins were identified. These included two monomeric (WMAI, four dimeric (WDAI, and six tetrameric (WTAI inhibitors of exogenous alpha-amylases, two inhibitors of endogenous alpha-amylases (WASI, four putative trypsin inhibitors (CMx and WTI, and one putative chymotrypsin inhibitor (WCI. A number of the encoded proteins were identical or very similar to proteins in the NCBI database. Sequences not reported previously included variants of WTAI-CM3, three CMx inhibitors and WTI. Within the WDAI group, two different genes encoded the same mature protein. Based on numbers of ESTs, transcripts for WTAI-CM3 Bu-1, WMAI Bu-1 and WTAI-CM16 Bu-1 were most abundant in Butte 86 developing grain. Coding sequences for 16 of the inhibitors were unequivocally associated with specific proteins identified by tandem mass spectrometry (MS/MS in a previous proteomic analysis of milled white flour from Butte 86. Proteins corresponding to WDAI Bu-1/Bu-2, WMAI Bu-1 and the WTAI subunits CM2 Bu-1, CM3 Bu-1 and CM16 Bu-1 were accumulated to the highest levels in flour. Conclusions Information on the spectrum of alpha-amylase/protease inhibitor genes and proteins expressed in a single wheat cultivar is central to understanding the importance of

  12. Effect of gelatinization and hydrolysis conditions on the selectivity of starch hydrolysis with alpha-amylase from Bacillus licheniformis

    NARCIS (Netherlands)

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

    2008-01-01

    Enzymatic hydrolysis of starch can be used to obtain various valuable hydrolyzates with different compositions. The effects of starch pretreatment, enzyme addition point, and hydrolysis conditions on the hydrolyzate composition and reaction rate during wheat starch hydrolysis with ¿-amylase from

  13. Degs and degu operon from Bacillus-brevis: a combination that enhances the production of commercially valuable enzymes

    CSIR Research Space (South Africa)

    Louw, M

    1995-05-01

    Full Text Available A novel method has been developed for increasing the production of commercially valuable enzymes, such as proteases, beta-glucanases, alpha-amylases and levansucrase. It is dependent on two genes cloned from Bacillus brevis, expressed on a multicopy...

  14. Reflection on design and testing of pancreatic alpha-amylase inhibitors: an in silico comparison between rat and rabbit enzyme models

    OpenAIRE

    Khalil-Moghaddam, Shiva; Ebrahim-Habibi, Azadeh; Pasalar, Parvin; Yaghmaei, Parichehreh; Hayati-Roodbari, Nasim

    2012-01-01

    Abstract Background Inhibitors of pancreatic alpha-amylase are potential drugs to treat diabetes and obesity. In order to find compounds that would be effective amylase inhibitors, in vitro and in vivo models are usually used. The accuracy of models is limited, but these tools are nonetheless valuable. In vitro models could be used in large screenings involving thousands of chemicals that are tested to find potential lead compounds. In vivo models are still used as preliminary mean of testing...

  15. RNA interference-based silencing of the alpha-amylase (amy1) gene in Aspergillus flavus decreases fungal growth and aflatoxin production in maize kernels.

    Science.gov (United States)

    Gilbert, Matthew K; Majumdar, Rajtilak; Rajasekaran, Kanniah; Chen, Zhi-Yuan; Wei, Qijian; Sickler, Christine M; Lebar, Matthew D; Cary, Jeffrey W; Frame, Bronwyn R; Wang, Kan

    2018-03-14

    Expressing an RNAi construct in maize kernels that targets the gene for alpha-amylase in Aspergillus flavus resulted in suppression of alpha-amylase (amy1) gene expression and decreased fungal growth during in situ infection resulting in decreased aflatoxin production. Aspergillus flavus is a saprophytic fungus and pathogen to several important food and feed crops, including maize. Once the fungus colonizes lipid-rich seed tissues, it has the potential to produce toxic secondary metabolites, the most dangerous of which is aflatoxin. The pre-harvest control of A. flavus contamination and aflatoxin production is an area of intense research, which includes breeding strategies, biological control, and the use of genetically-modified crops. Host-induced gene silencing, whereby the host crop produces siRNA molecules targeting crucial genes in the invading fungus and targeting the gene for degradation, has shown to be promising in its ability to inhibit fungal growth and decrease aflatoxin contamination. Here, we demonstrate that maize inbred B104 expressing an RNAi construct targeting the A. flavus alpha-amylase gene amy1 effectively reduces amy1 gene expression resulting in decreased fungal colonization and aflatoxin accumulation in kernels. This work contributes to the development of a promising technology for reducing the negative economic and health impacts of A. flavus growth and aflatoxin contamination in food and feed crops.

  16. Application of plackett-burman experimental design to optimize the cold-active alpha amylase biosynthesis by psychrotrophic Streptomyces 4 Alga

    Directory of Open Access Journals (Sweden)

    Mihaela COTÂRLEŢ

    2012-12-01

    Full Text Available The aim of this work was to optimize the cultural and production parameters through the statistical approach for the synthesis of cold active alpha amylase by Streptomyces 4 Alga in submerged fermentation. The process parameters influencing the enzyme production were identified using Plackett-Burman design. Among the various variables screened, the yeast extract and CaCl2 were the most significant. The optimum levels of these significant parameterswere determined employing the Response Surface Methodology and theCentral Composite Design. The most significant variables were determined as follows: yeast extract (5.00 g% and CaCl2 (0.25 g%. By using the optimal fermentation medium, the cold-active alpha amylase production was increased up to 23.96 AU, an approximate 4.4-fold improvement over the previous production (5.44 AU with un-optimized medium. The cold-active alpha amylases could be used in bioethanol production at lower temperatures, waste-water treatment and bioremediation in cold climates.

  17. Does potentized HgCl2 (Mercurius corrosivus) affect the activity of diastase and alpha-amylase?

    Science.gov (United States)

    Witt, Claudia M; Bluth, Michael; Hinderlich, Stephan; Albrecht, Henning; Lüdtke, Rainer; Weisshuhn, Thorolf E R; Willich, Stefan N

    2006-05-01

    Homeopathic drugs even with dilutions beyond 10(23) (high potencies) are frequently used, although their working mechanism is still unknown. Curative information preserved in solvent structure is postulated to exert biologic effects. The objective was to test for a stimulating or inhibiting effect of high potencies of the homeopathic remedy HgCl2 (Mercurius corrosivus) on two sugar hydrolases. High potencies were produced using stepwise dilution plus shaking. Controls included potentized solvent (aqua bidestillata), equimolar dilutions without shaking, and enzyme-free references. Tested were potencies with dilution factors 1:200 (CC) on diastase extract from winter barley, and 1:100 (C) on alpha-amylase from hog pancreas. Enzyme activity was colorimetrically determined by Lugol's iodine-starch reaction. An inhibiting effect of HgCl2 on enzyme activities was observed only in low potencies and dilutions. Statistically significant differences between potencies and controls were not found in randomized and blinded experiments. This experimental design provided independent reproducible results of cell-free in vitro assays. However, it did not indicate an effect of potentized HgCl2 on hydrolases. Demonstrating potency effects may require additional experimental features.

  18. In vitro and in vivo inhibition of alpha-amylases of stored-product mite Acarus siro.

    Science.gov (United States)

    Hubert, Jan; Dolecková-Maresová, Lucie; Hýblová, Jana; Kudlíková, Iva; Stejskal, Václav; Mares, Michael

    2005-01-01

    The stored-product mites are the most abundant and frequent group of pests living on the stored food products in Europe. They endanger public health since they produce allergens and transmit mycotoxin-producing fungi. Novel acaricidal compounds with inhibitory effects on the digestive enzymes of arthropods are a safe alternative to the traditional neurotoxic pesticides used for control of the stored-product pests. In this work, we explored the properties of acarbose, the low molecular weight inhibitor of alpha-amylases (AI), as a novel acaricide candidate for protection of the stored products from infestation by Acarus siro (Acari: Acaridae). In vitro analysis revealed that AI blocked efficiently the enzymatic activity of digestive amylases of A. siro, and decreased the physiological capacity of mite's gut in utilizing a starch component of grain flour. In vivo experiments showed that AI suppressed the population growth of A. siro. The mites were kept for three weeks on experimental diet enriched by AI in concentration range of 0.005 to 0.25%. Population growth of A. siro was negatively correlated with the content of AI in the treated diet with a half-population dose of 0.125%. The suppressive effect of AIs on stored-product mites is discussed in the context of their potential application in GMO crops.

  19. Biochemical and Structural Characterization of Amy1: An Alpha-Amylase from Cryptococcus flavus Expressed in Saccharomyces cerevisiae

    Directory of Open Access Journals (Sweden)

    Alexsandro Sobreira Galdino

    2011-01-01

    Full Text Available An extracellular alpha-amylase (Amy1 whose gene from Cryptococcus flavus was previously expressed in Saccharomyces cerevisiae was purified to homogeneity (67 kDa by ion-exchange and molecular exclusion chromatography. The enzyme was activated by NH4+ and inhibited by Cu+2 and Hg+2. Significant biochemical and structural discrepancies between wild-type and recombinant α-amylase with respect to Km values, enzyme specificity, and secondary structure content were found. Far-UV CD spectra analysis at pH 7.0 revealed the high thermal stability of both proteins and the difference in folding pattern of Amy1 compared with wild-type amylase from C. flavus, which reflected in decrease (10-fold of enzymatic activity of recombinant protein. Despite the differences, the highest activity of Amy1 towards soluble starch, amylopectin, and amylase, in contrast with the lowest activity of Amy1w, points to this protein as being of paramount biotechnological importance with many applications ranging from food industry to the production of biofuels.

  20. Interactions between salivary cortisol and alpha-amylase as predictors of children's cognitive functioning and academic performance.

    Science.gov (United States)

    Keller, Peggy S; El-Sheikh, Mona; Granger, Douglas A; Buckhalt, Joseph A

    2012-02-28

    We examined relations between salivary cortisol, alpha-amylase (sAA), and children's cognitive and academic functioning. Of interest were curvilinear and interactive effects of these salivary measures on cognitive and academic performance. Data were based on a sample of 28 boys and 36 girls (ages 8 and 9) in the Southeastern U.S.A. Children provided resting afternoon saliva samples. Children completed standardized tests of Intellectual Ability and schools provided academic achievement information. Regression analyses demonstrated significant curvilinear relations and interactions between cortisol and sAA in the prediction of child functioning. Contrary to current models of interactions among biological systems, findings indicated some of the highest and lowest scores were predicted at moderate levels of physiological arousal. For example, children with moderate sAA and either higher or lower cortisol had low predicted scores for Reading Ability. Children with moderate cortisol and lower sAA had the highest predicted scores for Intellectual Ability. Findings suggest that the study of interactions between biological stress response systems should not be based on models of rectilinear interactions. Copyright © 2011. Published by Elsevier Inc.

  1. Salivary cortisol and alpha-amylase: Is there consistency between psychosocial stress test and burdensome work shifts?

    Science.gov (United States)

    Karhula, Kati; Härmä, Mikko; Sallinen, Mikael; Lindholm, Harri; Hirvonen, Ari; Elovainio, Marko; Kivimäki, Mika; Vahtera, Jussi; Puttonen, Sampsa

    2017-12-01

    This study examined the consistency of salivary cortisol and alpha-amylase (sAA) total daily secretion between laboratory and field circumstances. The 95 participants were shift working female health care professionals with high (n = 53) or low (n = 42) psychosocial stress (job strain) measured by the Job Content Questionnaire (JCQ). The Trier Social Stress Test including a 5-min free speech and a mental arithmetic task was conducted with four, and field measurements with three daily saliva samples of cortisol and sAA during circadian rhythm and inter-shift recovery controlled morning shift, night shift, and a day off. The associations of salivary cortisol and sAA area under the curve with respect to ground (AUCg) and area under the curve with respect to increase (AUCi) between laboratory and field were tested using OLS (Ordinary Least Squares) regression. The sAA AUCg output in the laboratory was correlated with the output during all field measurement days and similarly among high and low job strain groups (p < 0.001). SAA AUCi and salivary cortisol AUCg and AUCi were not correlated between laboratory and field measurement, neither in the whole sample nor among the low or high job strain group. In conclusion, a laboratory measure of sAA AUCg output is promising in predicting stress-related output during burdensome work shifts and leisure time, whereas sAA AUCi or salivary cortisol seem not to have this potential.

  2. RNA interference and functional characterization of a tergal gland alpha amylase in the German cockroach, Blattella germanica L.

    Science.gov (United States)

    Myers, A J; Gondhalekar, A D; Fardisi, M; Pluchar, K D; Saltzmann, K D; Bennett, G W; Scharf, M E

    2018-04-01

    German cockroach males possess tergal glands that secrete a combination of oligosaccharides, lipids and proteins. Four major proteins occur in the secretion, with one being the 63 kDa alpha-amylase Blattella germanica Tergal Gland protein-1 (BGTG-1). Denaturing and starch gel electrophoresis coupled with peptide sequencing verified amylase activity for the BGTG-1 protein. BGTG-1 gene expression profiles were determined by using quantitative real-time PCR to compare messenger RNA abundance among isolated tissues of males, females and gravid females. Differences in BGTG-1 gene expression occurred among male tissues, with tergal gland tissue showing the highest expression. Tissues of nongravid and gravid females had significantly lower expression in comparison with male tergal glands (gravid females lowest). RNA interference (RNAi) was used to silence BGTG-1 gene expression by injecting BGTG-1 homologous double-stranded RNA (dsRNA) into male cockroaches. Groups injected with BGTG-1 dsRNA showed ∼90% lower BGTG-1 gene and protein expression compared to controls, which correlated with lower amylase activity in colorimetric assays. However, behavioural assays comparing precopulatory behaviour and mating success between RNAi and control males did not reveal differences. These results connect amylase gene expression and activity in tergal gland tissue but suggest other factors, such as other tergal gland components, may contribute more strongly to mating success. © 2017 The Royal Entomological Society.

  3. Effects of alpha-amylase reaction mechanisms on analysis of resistant-starch contents.

    Science.gov (United States)

    Moore, Samuel A; Ai, Yongfeng; Chang, Fengdan; Jane, Jay-lin

    2015-01-22

    This study aimed to understand differences in the resistant starch (RS) contents of native and modified starches obtained using two standard methods of RS content analysis: AOAC Method 991.43 and 2002.02. The largest differences were observed in native potato starch, cross-linked wheat distarch phosphate, and high-amylose corn starch stearic-acid complex (RS5) between using AOAC Method 991.43 with Bacillus licheniformis α-amylase (BL) and AOAC Method 2002.02 with porcine pancreatic α-amylase (PPA). To determine possible reasons for these differences, we hydrolyzed raw-starch granules with BL and PPA with equal activity at pH 6.9 and 37°C for up to 84 h and observed the starch granules displayed distinct morphological differences after the hydrolysis. Starches hydrolyzed by BL showed erosion on the surface of the granules; those hydrolyzed by PPA showed pitting on granule surfaces. These results suggested that enzyme reaction mechanisms, including the sizes of the binding sites and the reaction patterns of the two enzymes, contributed to the differences in the RS contents obtained using different methods of RS analysis. Copyright © 2014 Elsevier Ltd. All rights reserved.

  4. Improving the thermostability of alpha-amylase by combinatorial coevolving-site saturation mutagenesis

    Directory of Open Access Journals (Sweden)

    Wang Chenghua

    2012-10-01

    Full Text Available Abstract Background The generation of focused mutant libraries at hotspot residues is an important strategy in directed protein evolution. Existing methods, such as combinatorial active site testing and residual coupling analysis, depend primarily on the evolutionary conserved information to find the hotspot residues. Hardly any attention has been paid to another important functional and structural determinants, the functionally correlated variation information--coevolution. Results In this paper, we suggest a new method, named combinatorial coevolving-site saturation mutagenesis (CCSM, in which the functionally correlated variation sites of proteins are chosen as the hotspot sites to construct focused mutant libraries. The CCSM approach was used to improve the thermal stability of α-amylase from Bacillus subtilis CN7 (Amy7C. The results indicate that the CCSM can identify novel beneficial mutation sites, and enhance the thermal stability of wild-type Amy7C by 8°C (T5030, which could not be achieved with the ordinarily rational introduction of single or a double point mutation. Conclusions Our method is able to produce more thermostable mutant α-amylases with novel beneficial mutations at new sites. It is also verified that the coevolving sites can be used as the hotspots to construct focused mutant libraries in protein engineering. This study throws new light on the active researches of the molecular coevolution.

  5. Modulation of key metabolic enzyme of Labeo rohita (Hamilton) juvenile: effect of dietary starch type, protein level and exogenous alpha-amylase in the diet.

    Science.gov (United States)

    Kumar, Shivendra; Sahu, N P; Pal, A K; Sagar, Vidya; Sinha, Amit Kumar; Baruah, Kartik

    2009-06-01

    A 60-day feeding trial was conducted to delineate the effect of both gelatinized (G) and non-gelatinized (NG) corn with or without supplementation of exogenous alpha-amylase, either at optimum (35%) or sub-optimum (27%) protein levels, on blood glucose, and the key metabolic enzymes of glycolysis (hexokinase, HK), gluconeogenesis (glucose-6 phosphatase, G6Pase and fructose-1,6 bisphosphatase, FBPase), lipogenesis (glucose-6 phosphate dehydrogenase, G6PD) and amino acid metabolism (alanine amino transferase, ALT and aspartate amino transferase, AST) in Labeo rohita. Three hundred and sixty juveniles (average weight 10 +/- 0.15 g) were randomly distributed into 12 treatment groups with each of two replicates. Twelve semi-purified diets containing either 35 or 27% crude protein were prepared by including G or NG corn as carbohydrate source with different levels of microbial alpha-amylase (0, 50, 100 and 150 mg kg(-1)). The G corn fed groups showed significantly higher (P 0.05) on liver HK activity, but the optimum crude protein (35%) fed group showed higher HK activity than their low protein counterparts. The sub-optimum crude protein (27%) fed group showed significantly higher (P < 0.05) G6PD activity than the optimum protein fed group, whereas the reverse trend was observed for HK, G6Pase, FBPase, ALT and AST activity. Addition of 50 mg alpha-amylase kg(-1) feed showed increased blood glucose and G6PD activity of the NG corn fed group, whereas the reverse trend was found for G6Pase, FBPase, ALT and AST activity in liver, which was similar to that of the G or NG corn supplemented with 100/150 mg alpha-amylase kg(-1) feed. Data on enzyme activities suggest that NG corn in the diet significantly induced more gluconeogenic and amino acid metabolic enzyme activity, whereas G corn induced increased lipogenic enzyme activity. Increased amino acid catabolic enzyme (ALT and AST) activity was observed either at optimum protein (35%) irrespective of corn type or NG corn

  6. Structure-activity relationship of benzoxazinones and related compounds with respect to the growth inhibition and alpha-amylase activity in cress seedlings.

    Science.gov (United States)

    Kato-Noguchi, Hisashi; Macías, Francisco A; Molinillo, José M G

    2010-10-15

    Benzoxazinones and their degradation compounds inhibited root growth and alpha-amylase activity in cress seedlings. The inhibitory activity of these compounds was divided into three groups: the high active group; 2,4-dihydroxy-7-methoxy-(2H)-1,4-benzoxazin-3(4H)-one, 2,4-dihydroxy-(2H)-1,4-benzoxazin-3(4H)-one, 4-hydroxy-7-methoxy-(2H)-1,4-benzoxazin-3(4H)-one, 4-hydroxy-(2H)-1,4-benzoxazin-3(4H)-one, the moderate active group; 7-methoxy-(2H)-1,4-benzoxazin-3(4H)-one, (2H)-1,4-benzoxazin-3(4H)-one, 6-methoxy-benzoxazolin-2(3H)-one, benzoxazolin-2(3H)-one and 2-amino-phenoxazine-3-one, and the low active group; 2-hydroxy-(2H)-1,4-benzoxazin-3(4H)-one, 2-hydroxy-7-methoxy-(2H)-1,4-benzoxazin-3(4H)-one, 2-amino-7-hydroxyphenoxazine-3-one and 2-amino-7-methoxyphenoxazine-3-one. The structure-activity of these compounds suggests that compounds that have benzoxazinone skeletons are the most active structure, and a hydroxyl group at position C-2 on the benzoxazinone skeleton may not affect inhibitory activity, whereas a hydroxyl group at position N-4 on the skeleton is essential for inhibitory activity. However, the concentration-response curves of these compounds and the I(50) values (the concentrations required for 50% inhibition) for root growth and alpha-amylase indicated that root growth was positively correlated with the alpha-amylase activity in the seedlings. alpha-Amylase is required not only for seed germination, but also subsequent seedling growth until photosynthesis is sufficient to support seedling growth. Therefore, these results suggest that the compounds studied here may inhibit the root growth of cress seedlings by inhibiting alpha-amylase activity. Copyright (c) 2010 Elsevier GmbH. All rights reserved.

  7. Bacillus subtilis PrsA is required in vivo as an extracytoplasmic chaperone for secretion of active enzymes synthesized either with or without pro-sequences

    DEFF Research Database (Denmark)

    Jacobs, M; Kontinen, V; Sarvas, M

    1993-01-01

    In prsA (protein secretion) mutants of Bacillus subtilis, decreased levels of exoproteins, including alpha-amylase and subtilisins, are found extracellularly. The effect of prsA on subtilisin secretion is elaborated here. Extracytoplasmic folding and secretion of active subtilisin is assisted...

  8. Phylogenetic distribution of intron positions in alpha-amylase genes of bilateria suggests numerous gains and losses.

    Directory of Open Access Journals (Sweden)

    Jean-Luc Da Lage

    Full Text Available Most eukaryotes have at least some genes interrupted by introns. While it is well accepted that introns were already present at moderate density in the last eukaryote common ancestor, the conspicuous diversity of intron density among genomes suggests a complex evolutionary history, with marked differences between phyla. The question of the rates of intron gains and loss in the course of evolution and factors influencing them remains controversial. We have investigated a single gene family, alpha-amylase, in 55 species covering a variety of animal phyla. Comparison of intron positions across phyla suggests a complex history, with a likely ancestral intronless gene undergoing frequent intron loss and gain, leading to extant intron/exon structures that are highly variable, even among species from the same phylum. Because introns are known to play no regulatory role in this gene and there is no alternative splicing, the structural differences may be interpreted more easily: intron positions, sizes, losses or gains may be more likely related to factors linked to splicing mechanisms and requirements, and to recognition of introns and exons, or to more extrinsic factors, such as life cycle and population size. We have shown that intron losses outnumbered gains in recent periods, but that "resets" of intron positions occurred at the origin of several phyla, including vertebrates. Rates of gain and loss appear to be positively correlated. No phase preference was found. We also found evidence for parallel gains and for intron sliding. Presence of introns at given positions was correlated to a strong protosplice consensus sequence AG/G, which was much weaker in the absence of intron. In contrast, recent intron insertions were not associated with a specific sequence. In animal Amy genes, population size and generation time seem to have played only minor roles in shaping gene structures.

  9. Sex determines cortisol and alpha-amylase responses to acute physical and psychosocial stress in patients with avoidant personality disorder.

    Science.gov (United States)

    Tanaka, Yoshihiro; Ishitobi, Yoshinobu; Inoue, Ayako; Oshita, Harumi; Okamoto, Kana; Kawashima, Chiwa; Nakanishi, Mari; Aizawa, Saeko; Masuda, Koji; Maruyama, Yoshihiro; Higuma, Haruka; Kanehisa, Masayuki; Ninomiya, Taiga; Akiyoshi, Jotaro

    2016-08-01

    Avoidant personality disorder (AVPD) has excessive and pervasive anxiety and discomfort in social situations. The aims of this study were to explore the relationship between AVPD and physical and psychological stress and psychological tests. We evaluated 93 AVPD patients and 355 nonpatient controls by salivary amylase and cortisol responses during exposure to the Trier Social Stress Test (TSST) and electrical stimulation stress. Spielberger state-trait anxiety inventory (STAI), Profile of Mood State (POMS), Beck Depression Inventory (BDI), Depression and Anxiety Cognition Scale (DACS), and Childhood Trauma Questionnaire (CTQ) were administered. Following electrical stimulation, salivary cortisol levels in female AVPD decreased significantly less than that in female's controls, but salivary cortisol levels did not show a difference between male AVPD patients and controls. Salivary alpha-amylase (sAA) levels did not show a difference between females or male AVPD patients and controls. Following TSST exposure, sAA levels did not show a difference between females or male AVPD patients and controls. Salivary cortisol levels did not show a difference between females or male AVPD patients and controls. In the AVPD patients, POMS scores were significantly higher compared with the controls. STAI, BDI, DACS scores, and CTQ significantly increased in the AVPD patients compared with the controls. LF in heart rate variability in AVPD significantly increased more compared with controls. These results suggest that heightened sympathetic reactivity in female AVPD co-occurs with attenuated salivary cortisol responses to electric stimulation stress and there is a significant difference between AVPD and controls in mood, anxiety, social cognition, and automatic nerve systems.

  10. 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+}.

  11. Miconia sp. Increases mRNA Levels of PPAR Gamma and Inhibits Alpha Amylase and Alpha Glucosidase

    Directory of Open Access Journals (Sweden)

    David Mizael Ortíz-Martinez

    2016-01-01

    Full Text Available Diabetes mellitus is a public health problem worldwide. For this reason, ethanolic extract of Miconia sp. from Oaxaca, Mexico, was selected in search of an alternative against this disease. The effect of Miconia sp. on mRNA expression of PPARγ on cell line 3T3-L1, its effect on alpha amylase and alpha glucosidase, lipid accumulation during adipogenesis, and cell viability on VERO cells were evaluated. The mRNA levels of PPARγ increased on 1.393±0.008 folds, lipid accumulation was increased by 29.55% with Miconia sp. extract and 34.57% with rosiglitazone, and α-amylase and α-glycosidase were inhibited with IC50 values from 28.23±2.15 μg/mL and 1.95±0.15 μg/mL, respectively; the IC50 on antiproliferative activity on VERO cells was 314.54±45.40 μg/mL. In case of α-amylase and α-glycosidase assays, IC50 (inhibitory concentration 50 refers to necessary extract amounts to inhibit 50% of enzymatic activity. On the other hand, on antiproliferative activity, IC50 (inhibitory concentration 50 refers to necessary extract amounts to inhibit 50% of cell proliferation. It was concluded that the compounds present in Miconia sp. ethanolic extract increase mRNA expression of PPARγ, inhibit α-amylase and α-glucosidase, and increase lipid accumulation. It constitutes an alternative as adjuvant in diabetes mellitus treatment; therefore, we recommend continuing identifying the compounds responsible for its promising in vivo antidiabetic activity.

  12. Emotion regulation and positive affect in the context of salivary alpha-amylase response to pain in children with cancer.

    Science.gov (United States)

    Jenkins, Brooke N; Granger, Douglas A; Roemer, Ryan J; Martinez, Ariana; Torres, Tara K; Fortier, Michelle A

    2018-01-19

    Children with cancer routinely undergo painful medical procedures invoking strong physiological stress responses. Resilience to this pain may be conferred through resources such as emotion regulation strategies and positive affect. This study measured dispositional positive affect in children with cancer (N = 73) and randomly assigned participants to one of three emotion regulation strategy conditions (distraction, reappraisal, or reassurance). Children applied their assigned strategy during an experimental pain procedure (the cold pressor task [CPT]) and provided saliva samples before, immediately after, and 15 min after the CPT. Saliva samples were later assayed for salivary alpha amylase (sAA)-a surrogate marker for autonomic/sympathetic nervous system activity and regulation. Children in the reassurance group had sAA levels that continued to rise after completion of the CPT compared to children in the distraction (b = -1.68, P = 0.021) and reappraisal conditions (b = -1.24, P = 0.084). Furthermore, dispositional positive affect moderated the effect of condition such that children in the reassurance group with lower levels of positive affect had sAA levels that continued to rise after completion of the CPT (dy/dx = 1.56, P = 0.027), whereas children in the reassurance condition with higher levels of positive affect did not exhibit this rise (P > 0.05). Specific emotion regulation strategies, such as distraction and reappraisal, may attenuate the stress response to pain in pediatric patients with cancer, and positive affect may confer resilience in response to pain even with use of less effective coping strategies such as reassurance. © 2018 Wiley Periodicals, Inc.

  13. Differences in salivary alpha-amylase and cortisol responsiveness following exposure to electrical stimulation versus the Trier Social Stress Tests.

    Directory of Open Access Journals (Sweden)

    Yoshihiro Maruyama

    Full Text Available BACKGROUND: Cortisol is an essential hormone in the regulation of the stress response along the HPA axis, and salivary cortisol has been used as a measure of free circulating cortisol levels. Recently, salivary alpha-amylase (sAA has also emerged as a novel biomarker for psychosocial stress responsiveness within the sympathetic adrenomedullary (SAM system. PRINCIPAL FINDINGS: We measured sAA and salivary cortisol in healthy volunteers after exposure to the Trier Social Stress Test (TSST and electric stimulation stress. One hundred forty-nine healthy volunteers participated in this study. All subjects were exposed to both the TSST and electric stimulation stress on separate days. We measured sAA and salivary cortisol levels three times immediately before, immediately after, and 20 min after the stress challenge. The State (STAI-S and Trait (STAI-T versions of the Spielberger Anxiety Inventory test and the Profile of Mood State (POMS tests were administered to participants before the electrical stimulation and TSST protocols. We also measured HF, LF and LF/HF Heart Rate Variability ratio immediately after electrical stimulation and TSST exposure. Following TSST exposure or electrical stimulation, sAA levels displayed a rapid increase and recovery, returning to baseline levels 20 min after the stress challenge. Salivary cortisol responses showed a delayed increase, which remained significantly elevated from baseline levels 20 min after the stress challenge. Analyses revealed no differences between men and women with regard to their sAA response to the challenges (TSST or electric stimulations, while we found significantly higher salivary cortisol responses to the TSST in females. We also found that younger subjects tended to display higher sAA activity. Salivary cortisol levels were significantly correlated with the strength of the applied electrical stimulation. CONCLUSIONS: These preliminary results suggest that the HPA axis (but not the SAM

  14. Influence of the way of reporting alpha-Amylase values in saliva in different naturalistic situations: A pilot study.

    Directory of Open Access Journals (Sweden)

    María Dolores Contreras-Aguilar

    Full Text Available The objective of this pilot study was to compare the different ways of measuring salivary alpha-amylase (sAA, enzymatic vs. concentration and to evaluate the influence that the different ways of reporting the results can have in sAA interpretation. For this purpose, sAA was measured by direct quantification and also by an enzymatic assay in three different naturalistic situations, a physical stressor (situation 1 and two mental stressors of different intensity (situations 2 and 3. The results were expressed in three different ways (without correction, multiplied by flow rate and divided by protein concentration. sAA concentration and activity increased just after situations 1 and 3. When values were multiplied by the flow rate, significant changes after situation 1 were detected only for sAA activity but not for sAA concentration, being these changes of lower significance and magnitude that those observed for sAA activity without any correction. In addition, a significant increase in sAA activity was found at T+15 in situation 2. In situation 3 the significant decrease in sAA at T+15 disappeared. When values were divided by protein concentration, there were no significant changes in situations 1 or 3, but a decrease in situation 2 at T+0 and an increase at T+15. sAA activity and concentration showed a significant correlation in all situations. This pilot study points out that the way of expressing sAA can influence the results obtained in different stress models and also their interpretation. Therefore, how sAA is reported and the factors involved in the different ways of expressing sAA, should be taken into consideration for an objective interpretation of sAA values.

  15. Phylogenetic Distribution of Intron Positions in Alpha-Amylase Genes of Bilateria Suggests Numerous Gains and Losses

    Science.gov (United States)

    Da Lage, Jean-Luc; Maczkowiak, Frédérique; Cariou, Marie-Louise

    2011-01-01

    Most eukaryotes have at least some genes interrupted by introns. While it is well accepted that introns were already present at moderate density in the last eukaryote common ancestor, the conspicuous diversity of intron density among genomes suggests a complex evolutionary history, with marked differences between phyla. The question of the rates of intron gains and loss in the course of evolution and factors influencing them remains controversial. We have investigated a single gene family, alpha-amylase, in 55 species covering a variety of animal phyla. Comparison of intron positions across phyla suggests a complex history, with a likely ancestral intronless gene undergoing frequent intron loss and gain, leading to extant intron/exon structures that are highly variable, even among species from the same phylum. Because introns are known to play no regulatory role in this gene and there is no alternative splicing, the structural differences may be interpreted more easily: intron positions, sizes, losses or gains may be more likely related to factors linked to splicing mechanisms and requirements, and to recognition of introns and exons, or to more extrinsic factors, such as life cycle and population size. We have shown that intron losses outnumbered gains in recent periods, but that “resets” of intron positions occurred at the origin of several phyla, including vertebrates. Rates of gain and loss appear to be positively correlated. No phase preference was found. We also found evidence for parallel gains and for intron sliding. Presence of introns at given positions was correlated to a strong protosplice consensus sequence AG/G, which was much weaker in the absence of intron. In contrast, recent intron insertions were not associated with a specific sequence. In animal Amy genes, population size and generation time seem to have played only minor roles in shaping gene structures. PMID:21611157

  16. A proprietary alpha-amylase inhibitor from white bean (Phaseolus vulgaris: A review of clinical studies on weight loss and glycemic control

    Directory of Open Access Journals (Sweden)

    Barrett Marilyn L

    2011-03-01

    Full Text Available Abstract Obesity, and resultant health hazards which include diabetes, cardiovascular disease and metabolic syndrome, are worldwide medical problems. Control of diet and exercise are cornerstones of the management of excess weight. Foods with a low glycemic index may reduce the risk of diabetes and heart disease as well as their complications. As an alternative to a low glycemic index diet, there is a growing body of research into products that slow the absorption of carbohydrates through the inhibition of enzymes responsible for their digestion. These products include alpha-amylase and glucosidase inhibitors. The common white bean (Phaseolus vulgaris produces an alpha-amylase inhibitor, which has been characterized and tested in numerous clinical studies. A specific and proprietary product named Phase 2® Carb Controller (Pharmachem Laboratories, Kearny, NJ has demonstrated the ability to cause weight loss with doses of 500 to 3000 mg per day, in either a single dose or in divided doses. Clinical studies also show that Phase 2 has the ability to reduce the post-prandial spike in blood glucose levels. Experiments conducted incorporating Phase 2 into food and beverage products have found that it can be integrated into various products without losing activity or altering the appearance, texture or taste of the food. There have been no serious side effects reported following consumption of Phase 2. Gastro-intestinal side effects are rare and diminish upon extended use of the product. In summary, Phase 2 has the potential to induce weight loss and reduce spikes in blood sugar caused by carbohydrates through its alpha-amylase inhibiting activity.

  17. Combined effect of an antifeedant alpha-amylase inhibitor and a predator Cheyletus malaccensis in controlling the stored-product mite Acarus siro

    Czech Academy of Sciences Publication Activity Database

    Hubert, J.; Hýblová, Jana; Münzbergová, Zuzana; Pekár, S.; Křížková, I.; Marešová, Lucie; Stejskal, V.; Mareš, Michael

    2007-01-01

    Roč. 32, č. 1 (2007), s. 41-49 ISSN 0307-6962 R&D Projects: GA MŠk(CZ) 1P04OC842.20; GA MŠk(CZ) 1P04OC853.003; GA AV ČR IAA400550617; GA ČR GP203/02/P081 Institutional research plan: CEZ:AV0Z40550506; CEZ:AV0Z60050516 Keywords : alpha-amylase * Acarus siro * Cheyletus malaccensis * digestion protease Subject RIV: CC - Organic Chemistry Impact factor: 1.410, year: 2007

  18. Salivary cortisol and alpha-amylase diurnal profiles and stress reactivity in children with Attention Deficit Hyperactivity Disorder.

    Science.gov (United States)

    Angeli, Eleni; Korpa, Terpsichori; Johnson, Elizabeth O; Apostolakou, Filia; Papassotiriou, Ioannis; Chrousos, George P; Pervanidou, Panagiota

    2018-02-23

    There is growing evidence for dysregulation of the stress system in individuals with Attention Deficit Hyperactivity Disorder (ADHD). The stress system includes neuroanatomical and functional components that function in concert to maintain homeostasis and its main effectors are the hypothalamic-pituitary-adrenal (HPA) axis and the sympathetic/adrenomedullary nervous system (SNS). As stress system activity demonstrates a distinct circadian variation, we aimed to describe simultaneously, diurnal rhythms of both the HPA axis and the SNS in children with ADHD and a comparison group. Moreover, we attempted to investigate stress responses to a physical stressor, venipuncture, in both groups. Sixty-two prepubertal children with ADHD combined (ADHD-C) or inattentive (ADHD-I) type and 40 typically developing children provided saliva samples at six specific time points during a day, as well as before and 10 min after a scheduled morning venipuncture. Salivary cortisol and α- amylase were selected as reliable noninvasive biomarkers for HPA axis and SNS function and were measured in the samples obtained. Results revealed that children with ADHD-C had lower mean cortisol values both 30 min after awakening and at 18:00 h than controls (p = 0.002 and p = 0.018 respectively), as well as lower mean Cortisol Awakening Response (CAR) and Area Under the Curve for "wake to bed" period (AUC i ) values of cortisol (p = 0.004 and p = 0.001, respectively). Also, mean CAR and cortisol AUC i were lower in children with ADHD-I than the control group (p = 0.034 and p = 0.038 respectively). Alpha-amylase measurements showed an increase over time (p < 0.001), which was similar in all three groups. Interestingly, α-amylase changes over time were correlated with the corresponding cortisol changes (p < 0.001). Venipuncture, elicited a significant increase only in α-amylase levels and more so in the control group (p = 0.003). These findings suggest a

  19. Alpha-Amylase

    Science.gov (United States)

    2004-01-01

    Both (Porcine and bacterial) starch degrading enzymes highly valued by the biotechnology industry. (Porcine) A major target for protein engineering and the study of diabetes, obesity and dental care. (Bacterial) Major industrial and biotechnology interest used in brewing, baking, and food processing. World's number one industrial protein.

  20. A stochastic model for predicting dextrose equivalent and saccharide composition during hydrolysis of starch by alpha-amylase

    NARCIS (Netherlands)

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

    2008-01-01

    A stochastic model was developed that was used to describe the formation and breakdown of all saccharides involved during -amylolytic starch hydrolysis in time. This model is based on the subsite maps found in literature for Bacillus amyloliquefaciens -amylase (BAA) and Bacillus licheniformis

  1. Starch digestion in tropical fishes: isolation, structural studies and inhibition kinetics of alpha-amylases from two tilapias Oreochromis niloticus and Sarotherodon melanotheron.

    Science.gov (United States)

    Moreau, Y; Desseaux, V; Koukiekolo, R; Marchis-Mouren, G; Santimone, M

    2001-03-01

    alpha-Amylases from the intestinal cavity of two tilapia species, Oreochromis niloticus (ONI-AMY) and Sarotherodon melanotheron (SME-AMY), were purified using ammonium sulfate precipitation, affinity chromatography and chromatofocusing procedures. The purification was approximately 100-fold. The amylolytic activity, specific activity, product distribution, pH and temperature profile of ONI-AMY and SME-AMY are quite similar. The molecular mass differs slightly: 56600 (ONI-AMY) vs. 55500 (SME-AMY). As shown by isoelectric focusing analysis, both amylases contain two isoforms A and B with distinct pI: 7.2 (A) and 7.8 (B), vs. 8.3 (A) and 8.8 (B), respectively. It was not possible to isolate B, since B converts into A with time. The kinetics of the inhibition of ONI-AMY and SME-AMY activity by alpha-, beta- and gamma-cyclodextrin (alpha-, beta- and gamma-CD) were investigated using amylose as the substrate. Statistical analysis of the kinetic data expressed using a general velocity equation and assuming rapid equilibrium showed that the inhibition is of the mixed noncompetitive type. Similar results were obtained with ONI-AMY and SME-AMY. beta- and gamma-CD are stronger inhibitors than alpha-CD. ONI-AMY and SME-AMY are then closely related and show the general features common to the members of the alpha-amylase class (family 13). They enable ONI and SME tilapias to digest starch in food.

  2. Direct production of L-lysine from raw corn starch by Corynebacterium glutamicum secreting Streptococcus bovis alpha-amylase using cspB promoter and signal sequence.

    Science.gov (United States)

    Tateno, Toshihiro; Fukuda, Hideki; Kondo, Akihiko

    2007-12-01

    Corynebacterium glutamicum is an important microorganism in the industrial production of amino acids. We engineered a strain of C. glutamicum that secretes alpha-amylase from Streptococcus bovis 148 (AmyA) for the efficient utilization of raw starch. Among the promoters and signal sequences tested, those of cspB from C. glutamicum possessed the highest expression level. The fusion gene was introduced into the homoserine dehydrogenase gene locus on the chromosome by homologous recombination. L-Lysine fermentation was conducted using C. glutamicum secreting AmyA in the growth medium containing 50 g/l of raw corn starch as the sole carbon source at various temperatures in the range 30 to 40 degrees C. Efficient L-lysine production and raw starch degradation were achieved at 34 and 37 degrees C, respectively. The alpha-amylase activity using raw corn starch was more than 2.5 times higher than that using glucose as the sole carbon source during L-lysine fermentation. AmyA expression under the control of cspB promoter was assumed to be induced when raw starch was used as the sole carbon source. These results indicate that efficient simultaneous saccharification and fermentation of raw corn starch to L-lysine were achieved by C. glutamicum secreting AmyA using the cspB promoter and signal sequence.

  3. Localization of an O-glycosylated site in the recombinant barley alpha-amylase 1 produced in yeast and correction of the amino acid sequence using matrix-assisted laser desorption/ionization mass spectrometry of peptide mixtures

    DEFF Research Database (Denmark)

    Andersen, Jens S.; Søgaard, M; Svensson, B

    1994-01-01

    Matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) of peptide mixtures was used to characterize recombinant barley alpha-amylase 1, produced in yeast. Three peptide mixtures were generated by cleavage with CNBr, digestion with endoproteinase Lys-C and Asp-N, respectively...

  4. 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.

  5. 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.

  6. Mapping of barley alpha-amylases and outer subsite mutants reveals dynamic high-affinity subsites and barriers in the long substrate binding cleft

    DEFF Research Database (Denmark)

    Kandra, L.; Abou Hachem, Maher; Gyemant, G.

    2006-01-01

    Subsite affinity maps of long substrate binding clefts in barley alpha-amylases, obtained using a series of maltooligosaccharides of degree of polymerization of 3-12, revealed unfavorable binding energies at the internal subsites -3 and -5 and at subsites -8 and +3/+4 defining these subsites...... as binding barriers. Barley a-amylase I mutants Y105A and T212Y at subsite -6 and +4 resulted in release or anchoring of bound substrate, thus modifying the affinities of other high-affinity subsites (-2 and +2) and barriers. The double mutant Y105A-T212Y displayed a hybrid subsite affinity profile......, converting barriers to binding areas. These findings highlight the dynamic binding energy distribution and the versatility of long maltooligosaccharide derivatives in mapping extended binding clefts in a-amylases....

  7. Stress exacerbates pain in the everyday lives of women with fibromyalgia syndrome--The role of cortisol and alpha-amylase.

    Science.gov (United States)

    Fischer, Susanne; Doerr, Johanna M; Strahler, Jana; Mewes, Ricarda; Thieme, Kati; Nater, Urs M

    2016-01-01

    Although fibromyalgia syndrome (FMS) is a chronic condition, its cardinal symptom pain is known to fluctuate over the day. Stress has often been claimed to exacerbate pain; however, there is barely any evidence on whether or not this is true on a day-to-day basis (and, alternatively, on whether pain leads to increased stress levels). Using an ecologically valid measurement design, we tested whether and how stress and pain are intertwined in participants with FMS. We additionally examined the role of the two major stress-responsive systems, the hypothalamic-pituitary-adrenal axis and the autonomic nervous system, as potential mediators of this relationship. An ambulatory assessment study was conducted over the course of 14 days. On each day, 32 females with FMS provided six diary entries on momentary stress and pain levels. Saliva samples were collected at the same time points to determine cortisol and alpha-amylase as indicators of stress-responsive systems. Higher stress at a given measurement time point was associated with higher reported pain levels at the subsequent time point (UC=1.47, ppain relationship was neither mediated by momentary cortisol nor by alpha-amylase; however, momentary cortisol was independently associated with momentary pain (UC=0.27, p=0.009). Stress seems to be a powerful exacerbating factor for pain as experienced by patients with FMS in their everyday lives. Cortisol may be involved in the diurnal fluctuation of pain levels in patients with FMS. Future studies should identify relevant daily stressors in persons with FMS and scrutinize the mechanisms underlying the cortisol-pain relationship. Copyright © 2015 Elsevier Ltd. All rights reserved.

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

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    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.

  9. Salivary cortisol, alpha-amylase and immunoglobulin a responses to a morning session of basketball or volleyball training in boys aged 14-18 years.

    Science.gov (United States)

    Bruzda-Zwiech, A; Konieczka, M; Hilt, A; Daszkowska, M; Grzegorczyk, J; Szczepańska, J

    2017-01-01

    This study investigates whether a single session of routine morning basketball or volleyball training affects saliva levels of cortisol, alpha-amylase (sAA) and secretory immunoglobulin A (sIgA) in boys aged 14–18 years. Twenty-nine boys who participate in basketball or volleyball training, recruited from the Marcin Gortat’s Athletic Championship School in Lodz, were enrolled in the study. The 90-minute routine exercise program included 15 minutes of warm-up followed by basketball or volleyball practice. Unstimulated saliva samples were collected prior to and immediately after the exercise, and were analysed using ELISA. One training session resulted in a significant increase of sAA concentration in all participants, as well as in the volleyball and basketball subgroups (p=0.00022; p=0.0029; p=0.0011; respectively). Post-exercise cortisol levels were significantly lower than pre-exercise levels (p=0.00002) throughout the group, as well as in the volleyball and basketball subgroups (p=0.0048; p=0.0019; p=0.0048; respectively). The exercise protocol did not significantly affect sIgA level, either in the whole examined group or the volleyball subgroup, however a weak significant increase of sIgA was observed in the basketball subgroup (p=0.046). The routine morning training session comprising a warm-up followed by basketball or volleyball practice seems to activate the sympatho-adrenal-medullary system, with a subsequent increase of alpha-amylase, but does not affect oral immunity in 14-18-year-old boys.

  10. Salivary cortisol and alpha-amylase levels during an assessment procedure correlate differently with risk-taking measures in male and female police recruits

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    Ruud eVan Den Bos

    2014-01-01

    Full Text Available Recent laboratory studies have shown that men display more risk-taking behaviour in decision-making tasks following stress, whilst women are more risk-aversive or become more task-focussed. In addition, these studies have shown that sex differences are related to levels of the stress hormone cortisol (indicative of activation of the hypothalamus-pituitary-adrenocortical-axis: the higher the levels of cortisol the more risk-taking behaviour is shown by men, whereas women generally display more risk-aversive or task-focussed behaviour following higher levels of cortisol. Here, we assessed whether such relationships hold outside the laboratory, correlating levels of cortisol obtained during a job-related assessment procedure with decision-making parameters in the Cambridge Gambling Task (CGT in male and female police recruits. The CGT allows for discriminating different aspects of reward-based decision-making. In addition, we correlated levels of alpha-amylase (indicative for activation of the sympatho-adrenomedullary-axis and decision-making parameters. In line with earlier studies men and women only differed in risk-adjustment in the CGT. Salivary cortisol levels correlated positively and strongly with risk-taking measures in men, which was significantly different from the weak negative correlation in women. In contrast, and less strongly so, salivary alpha-amylase levels correlated positively with risk-taking in women, which was significantly different from the weak negative correlation with risk-taking in men. Collectively, these data support and extend data of earlier studies indicating that risky decision-making in men and women is differently affected by stress hormones. The data are briefly discussed in relation to the effects of stress on gambling.

  11. Bacillus catenulatus sp. nov., an alkalitolerant bacterium isolated from a soda lake.

    Science.gov (United States)

    Sultanpuram, Vishnuvardhan Reddy; Mothe, Thirumala; Chintalapati, Sasikala; Chintalapati, Venkata Ramana

    2017-12-01

    Two novel (18C T and 6C) Gram-stain-positive, rod shaped, motile and endospore-forming bacterial strains were isolated from Lonar soda lake, India. Based on 16S rRNA gene sequence analysis, strains 18C T and 6C were identified as belonging to the class Firmibacteria, and were most closely related to Bacillus cohnii KCTC 3572 T (99.3 and 99.9%, respectively), Bacillus zhanjiangensis KCTC 13713 T (97.4 and 98.0%, respectively), Bacillus halmapalus LMG 17950 T (97.0 and 97.6%, respectively) and other members in the genus Bacillus (Bacillus, for which the name Bacillus catenulatus sp. nov. is proposed. The type strain is 18C T (=KCTC 33781 T  = CGMCC 1.15475 T ).

  12. [Effect of citric acid stimulation on salivary alpha-amylase, total protein, salivary flow rate and pH value in Pi deficiency children].

    Science.gov (United States)

    Yang, Ze-min; Chen, Long-hui; Lin, Jing; Zhang, Min; Yang, Xiao-rong; Chen, Wei-wen

    2015-02-01

    To compare the effect of citric acid stimulation on salivary alpha-amylase (sAA), total protein (TP), salivary flow rate, and pH value between Pi deficiency (PD) children and healthy children, thereby providing evidence for Pi controlling saliva theory. Twenty PD children were recruited, and 29 healthy children were also recruited at the same time. Saliva samples from all subjects were collected before and after citric acid stimulation. The sAA activity and amount, TP contents, salivary flow rate, and pH value were determined and compared. (1) Citric acid stimulation was able to significantly increase salivary flow rate, pH value, sAA activities, sAA specific activity and sAA amount (including glycosylated and non-glycosylated sAA amount) in healthy children (Pvalue, and glycosylated sAA levels in PD children (P0.05), salivary indices except salivary flow rate and glycosylated sAA levels decreased more in PD children. There was statistical difference in sAA activity ratio, sAA specific activity ratio, and the ratio of glycosylated sAA levels between PD children and healthy children (P<0.05). PD children had decreased response to citric acid stimulation.

  13. Mind your thoughts: associations between self-generated thoughts and stress-induced and baseline levels of cortisol and alpha-amylase.

    Science.gov (United States)

    Engert, Veronika; Smallwood, Jonathan; Singer, Tania

    2014-12-01

    Stress is a major health burden in today's society. Research shows that negative cognitive styles are associated with increased stress reactivity, low mood and accelerated cellular aging. Our study sought to unravel the relationship between the content of self-generated thoughts and psychosocial stress measured in terms of hypothalamic-pituitary-adrenal axis and sympathetic activity. Features of self-generated thoughts were assessed using thought sampling while participants performed cognitive tasks following a stress induction or in a baseline condition. More negatively toned emotional thoughts and more social temporal thoughts with a past focus were associated with increased cortisol and alpha-amylase levels, both after stress and at baseline. More social temporal thoughts with a future focus, on the other hand, had an overall attenuating effect on the levels of both stress markers. Our results indicate a fundamental link between the thoughts and stress levels we experience. Understanding the mechanisms governing this mind-body association may have important implications for understanding and counteracting the high incidence of stress-related disorders in today's society. Copyright © 2014 Elsevier B.V. All rights reserved.

  14. Effect of domestic cooking on the starch digestibility, predicted glycemic indices, polyphenol contents and alpha amylase inhibitory properties of beans (Phaseolis vulgaris) and breadfruit (Treculia africana).

    Science.gov (United States)

    Chinedum, E; Sanni, S; Theressa, N; Ebere, A

    2018-01-01

    The effect of processing on starch digestibility, predicted glycemic indices (pGI), polyphenol contents and alpha amylase inhibitory properties of beans (Phaseolis vulgaris) and breadfruit (Treculia africana) was studied. Total starch ranged from 4.3 to 68.3g/100g, digestible starch ranged from 4.3 to 59.2 to 65.7g/100g for the raw and processed legumes; Resistance starch was not detected in most of the legumes except in fried breadfruit and the starches in both the raw and processed breadfruit were more rapidly digested than those from raw and cooked beans. Raw and processed breadfruit had higher hydrolysis curves than raw and processed beans with the amylolysis level in raw breadfruit close to that of white bread. Raw beans had a low glycemic index (GI); boiled beans and breadfruit had intermediate glycemic indices respectively while raw and fried breadfruit had high glycemic indices. Aqueous extracts of the food samples had weak α-amylase inhibition compared to acarbose. The raw and processed legumes contained considerable amounts of dietary phenols and flavonoids. The significant correlation (r=0.626) between α-amylase inhibitory actions of the legumes versus their total phenolic contents suggests the contribution of the phenolic compounds in these legumes to their α-amylase inhibitory properties. Copyright © 2017 Elsevier B.V. All rights reserved.

  15. Effects of a high-pressure treatment on the wheat alpha-amylase inhibitor and its relationship to elimination of allergenicity

    Science.gov (United States)

    Yamamoto, S.; Takanohashi, K.; Hara, T.; Odani, S.; Suzuki, A.; Nishiumi, T.

    2010-03-01

    In this study, the effects of high-pressure treatment on structure and allergeincity of alpha amylase inhibitor (a-AI) were investigated. The pressure-induced structural changes of α-AI were estimated by fluorescence spectra and by fourth derivative UV-spectroscopy for probed tyrosine residues and by circular dichroism (CD) spectroscopy. The changes in the tertiary structure detected by fluorescence spectra and by fourth derivative UV-spectroscopy under high pressure were indicated at over 300 MPa. Measurements of CD spectroscopy suggested that the effects of a high-pressure treatment on changes in the secondary structure of α-AI were little. From our results, pressure-induced changes of the α-AI structure were not apparent. On the other hands, the IgE-specific binding activities of pressurized α-AI to sera from allergic patients against wheat, which is estimated by observations of dot-blotting, were decreased by high-pressure treatment. It is known that the pressure-induced elimination of allergenicity is related to the tertiary structural changes of allergen molecules. This study are suspected that the epitopes of α-AI do not contain tyrosine residues, and thus the decrease of IgE-specific binding activities is probably caused by the tertiary structural changes of these parts of α-AI.

  16. In vitro alpha-amylase inhibition and in vivo antioxidant potential of Momordica dioica seeds in streptozotocin-induced oxidative stress in diabetic rats

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    P. Sailaja Rao

    2017-09-01

    Full Text Available Momordica dioica Roxb. Commonly known as “Kakora” in Telugu, is used in the Indian traditional system of medicine for the treatment of diabetes. The aim of this study was to investigate the antidiabetic activity of methanolic extract of M. dioica seeds (MEMD in streptozotocin (STZ induced diabetic rats. The in vitro α-amylase inhibitory activity of the MEMD was done by spectrophotometric method. Diabetes was induced by STZ (45 mg/kg; i.p, MEMD (100 & 200 mg/kg; b.wt and standard drug metformin (50 mg/kg; b.wt were administered to the diabetic rats. Blood glucose was estimated on the 11th day and the level of MDA, SOD and CAT was estimated in the liver tissue homogenate after the 15 days of experimental period. MEMD showed significant inhibition of alpha amylase activity and the IC50 was found to be 48 μg/ml. Oral administration of MEMD significantly reduced blood glucose level (P < 0.05, diminished the MDA level and refurbished depleted antioxidant enzymes and Insulin level to normalcy. These findings revealed that M. dioica seeds possess antihyperglycemic, antioxidant and anti lipid peroxidative activity and thus mitigate STZ-induced oxidative damage.

  17. Effects of a high-pressure treatment on the wheat alpha-amylase inhibitor and its relationship to elimination of allergenicity

    Energy Technology Data Exchange (ETDEWEB)

    Yamamoto, S [Food Science Center, Niigata University, Ikarashi, Niigata, 950-2181 (Japan); Takanohashi, K; Nishiumi, T [Graduate School of Science and Technology, Niigata University, Ikarashi, Niigata, 950-2181 (Japan); Hara, T [Department of Applied Biological Chemistry, Faculty of Agriculture, Niigata University, Ikarashi, Niigata, 950-2181 (Japan); Odani, S [Department of Living Science and Technology, Faculty of Education and Human Science, Ikarashi, Niigata, 950-2181 (Japan); Suzuki, A, E-mail: shuyama@agr.niigata-u.ac.j [Department of Health and Nutrition, Faculty of Medical Science for Health, Teikyo Heisei University, Ikebukuro, Tokyo, 170-0013 (Japan)

    2010-03-01

    In this study, the effects of high-pressure treatment on structure and allergeincity of alpha amylase inhibitor (a-AI) were investigated. The pressure-induced structural changes of {alpha}-AI were estimated by fluorescence spectra and by fourth derivative UV-spectroscopy for probed tyrosine residues and by circular dichroism (CD) spectroscopy. The changes in the tertiary structure detected by fluorescence spectra and by fourth derivative UV-spectroscopy under high pressure were indicated at over 300 MPa. Measurements of CD spectroscopy suggested that the effects of a high-pressure treatment on changes in the secondary structure of {alpha}-AI were little. From our results, pressure-induced changes of the {alpha}-AI structure were not apparent. On the other hands, the IgE-specific binding activities of pressurized {alpha}-AI to sera from allergic patients against wheat, which is estimated by observations of dot-blotting, were decreased by high-pressure treatment. It is known that the pressure-induced elimination of allergenicity is related to the tertiary structural changes of allergen molecules. This study are suspected that the epitopes of {alpha}-AI do not contain tyrosine residues, and thus the decrease of IgE-specific binding activities is probably caused by the tertiary structural changes of these parts of {alpha}-AI.

  18. May Salivary Alpha-Amylase Level Be a Useful Tool for Assessment of the Severity of Schizophrenia and Evaluation of Therapy? A Case Report

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    Masa Ieda

    2012-01-01

    Full Text Available Background. Previous studies suggested dysfunction of the autonomic nervous system (ANS in schizophrenia patients, but the mechanism remains unclear. Recently, the measurement of salivary alpha-amylase (sAA has been considered a useful tool for evaluating ANS, especially the sympathoadrenal medullary system. Furthermore, there was a report that patients with schizophrenia showed much higher sAA level than normal controls. Methods. We present the case of a 51-year-old female with catatonic schizophrenia. She needed the treatment of electroconvulsive therapy (ECT. We evaluated her sAA level and her psychiatric symptoms during the treatment. Results. Before ECT treatment, she showed high sAA level. Her sAA level decreased during the course of ECT, and this attenuation was accompanied by improvement of schizophrenic symptoms. Conclusion. We consider that measurement of the sAA level may be one of the useful biological markers for assessment of psychotic state and efficacy of treatment in patients with schizophrenia.

  19. Refining the multisystem view of the stress response: coordination among cortisol, alpha-amylase, and subjective stress in response to relationship conflict.

    Science.gov (United States)

    Laurent, Heidemarie K; Powers, Sally I; Granger, Douglas A

    2013-07-02

    This study investigated associations among young adults' hypothalamic-pituitary-adrenal axis activity, autonomic nervous system activity, and subjective stress in response to interpersonal conflict to better characterize coordination across stress systems. Seven saliva samples were collected from 199 young adult opposite-sex couples before, during, and after they discussed an unresolved relationship conflict. Samples were later assayed for cortisol and alpha-amylase (sAA). Couples rated anticipatory stress prior to the conflict and perceived stress immediately following the task. Growth curve modeling was used to examine two possible levels of within-person coordination across physiological systems: alignment between cortisol and sAA responses throughout the sampling period ("matched phase coordination"), and association between overall levels of cortisol and sAA in response to conflict ("average level coordination"). Whereas both partners showed the former type of coordination, only women showed the latter type. Positive anticipation of the stressor predicted stronger cortisol-sAA matched phase coordination for women. Pre-task ratings related to women's sAA, and post-task ratings related to both partners' cortisol responses. Implications for a multisystem interpretation of normal and pathological responses to daily stress are discussed. Copyright © 2013 Elsevier Inc. All rights reserved.

  20. Dietary Intake, Eating Behaviors, and Diurnal Patterns of Salivary Cortisol and Alpha-Amylase Secretion Among Professional Young Adult Female Tennis Players.

    Science.gov (United States)

    Filaire, Edith; Massart, Alain; Hua, Jiewen; Le Scanff, Christine

    2015-06-01

    The aims of study were to examine the eating behaviors among 26 professional female tennis players and to assess the diurnal patterns of stress hormones through the measurement of awakening and diurnal profiles of salivary alpha-amylase (sAA) and cortisol concentrations. Eating behaviors were assessed through three questionnaires (Eating Attitudes Test-26; Eating Disorders Inventory 2; and Body Shape Questionnaire), food intake by a 7-day diet record, and menstrual status by questionnaire. Perceived stress scale and anxiety state were also evaluated. Saliva samples were collected at awakening, 30 min, 60 min, and 12 hr post awakening after 6-days' rest. Forty-six percent of tennis players presented Disordered Eating attitudes (DE) (n = 12) with a lower body mass index, and higher state anxiety as compared with the group without DE. No differences in the Perceived Stress Scale scores were noted. Mean energy intake, protein and carbohydrates intakes were lower (p consequences of these modifications on health remain to be elucidated.

  1. Use of alpha-amylase and amyloglucosidase combinations to minimize the bread quality problems caused by high levels of damaged starch.

    Science.gov (United States)

    Barrera, Gabriela N; Tadini, Carmen C; León, Alberto E; Ribotta, Pablo D

    2016-10-01

    The objective of this work was to investigate the contribution of α-amylase and amyloglucosidase to dough fermentation process and bread quality, as an alternative to reduce the negative effects caused by high damaged starch in flour. The dough properties during the proofing process were modified by higher damaged starch content. Higher damaged starch in flour resulted into breads with darker crusts and firmer crumbs. The enzymes reduced the negative influence of damaged starch, producing a positive effect on the maximum carbon dioxide pressure reached during fermentation and the carbon dioxide volume retained by dough. Incorporation of alpha-amylase reduced dimension ratio and crumb firmness attributes; however, progressive additions of this additive produced lower bread volume and red intensity, and higher crumb firmness. The amyloglucosidase additions produced higher bread volume and red intensity of the crust, and lower brightness crust and gas cell diameter. Incorporation of amyloglucosidase was beneficial in the presence of a suitable quantity of damaged starch. The results confirmed that the α-amylase and amyloglucosidase additions significantly improved bread quality. Incorporation of α-amylase and amyloglucosidase led to higher bread loaves and lower crumb firmness throughout the storage period, promoting a longer life of the finished product.

  2. Person-centered examination of salivary cortisol and alpha-amylase responses to psychosocial stress: Links to preadolescent behavioral functioning and coping.

    Science.gov (United States)

    Bendezú, Jason José; Wadsworth, Martha E

    2018-02-01

    This study adopted a person-centered approach to identify preadolescent salivary cortisol (sC) and alpha-amylase (sAA) co-activation response patterns and examine links to behavioral functioning and coping. Children (N = 151, 51.7% male) were exposed to the Trier Social Stress Test (TSST) and one of two randomly-assigned, post-TSST coping conditions: distraction or avoidance. Multi-trajectory modeling yielded four child subgroups. Child internalizing and externalizing positively predicted High sC-High sAA relative to Low sC-Low sAA and Low sC-High sAA relative to High sC-Low sAA subgroup membership, respectively. Low sC-Low sAA children demonstrated more efficient sC recovery when primed with distraction and more protracted sC recovery when primed with avoidance. For High sC-High sAA, internalizing children, the opposite was true. Findings illustrate adjustment-linked variability in preadolescent sC-sAA co-activation response patterns that further articulates for whom effortful coping works to effectively manage stressor-induced neuroendocrine activation. Copyright © 2017 Elsevier B.V. All rights reserved.

  3. Effects of a high-pressure treatment on the wheat alpha-amylase inhibitor and its relationship to elimination of allergenicity

    International Nuclear Information System (INIS)

    Yamamoto, S; Takanohashi, K; Nishiumi, T; Hara, T; Odani, S; Suzuki, A

    2010-01-01

    In this study, the effects of high-pressure treatment on structure and allergeincity of alpha amylase inhibitor (a-AI) were investigated. The pressure-induced structural changes of α-AI were estimated by fluorescence spectra and by fourth derivative UV-spectroscopy for probed tyrosine residues and by circular dichroism (CD) spectroscopy. The changes in the tertiary structure detected by fluorescence spectra and by fourth derivative UV-spectroscopy under high pressure were indicated at over 300 MPa. Measurements of CD spectroscopy suggested that the effects of a high-pressure treatment on changes in the secondary structure of α-AI were little. From our results, pressure-induced changes of the α-AI structure were not apparent. On the other hands, the IgE-specific binding activities of pressurized α-AI to sera from allergic patients against wheat, which is estimated by observations of dot-blotting, were decreased by high-pressure treatment. It is known that the pressure-induced elimination of allergenicity is related to the tertiary structural changes of allergen molecules. This study are suspected that the epitopes of α-AI do not contain tyrosine residues, and thus the decrease of IgE-specific binding activities is probably caused by the tertiary structural changes of these parts of α-AI.

  4. 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.)

  5. 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.

  6. Salivary alpha amylase diurnal pattern and stress response are associated with body mass index in low-income preschool-aged children.

    Science.gov (United States)

    Miller, Alison L; Sturza, Julie; Rosenblum, Katherine; Vazquez, Delia M; Kaciroti, Niko; Lumeng, Julie C

    2015-03-01

    Physiological stress responses are proposed as a pathway through which stress can "get under the skin" and lead to health problems, specifically obesity. We tested associations of salivary alpha amylase (sAA) diurnal patterns and stress responses with body mass index (BMI) in young, low-income children (51% male; 54% non-Hispanic white). Diurnal saliva samples were collected three times per day across three days for 269 children (M age 50.8 months, SD 6.3). Individual sAA intercept and slope values were calculated using random effect models to represent morning sAA levels and rate of sAA change across the day. A subset of children (n=195; M age 56.6 months, SD 6.9) participated in a lab-based behavioral stress protocol. Area under the curve increase (AUCI) across four timepoints was calculated to represent increase in sAA output during stress elicitation. Children were weighed and height measured and BMI z-score was calculated. Linear regression was used to evaluate associations of sAA intercept, sAA slope, and sAA AUCI with BMI z-score, controlling for child age, sex, and race/ethnicity; maternal weight status; and family income-to-needs ratio. Diurnal and stress-response sAA patterns were related to child adiposity: for each 1-standard deviation unit (SDU) decrease in morning sAA level, the child's BMI z-score increased by 0.11 (SE 0.05) SDU's (pstress elicitation, the child's BMI z-score increased by 0.14 (SE 0.06) SDU's (pstress responses and atypical diurnal patterns of sAA have been found following exposure to chronic life stressors such as poverty. Findings suggest that associations of stress, sAA, and elevated body mass index may develop very early in the lifespan. Copyright © 2014 Elsevier Ltd. All rights reserved.

  7. Measurements of salivary alpha amylase and salivary cortisol in hominoid primates reveal within-species consistency and between-species differences.

    Directory of Open Access Journals (Sweden)

    Verena Behringer

    Full Text Available Salivary alpha amylase (sAA is the most abundant enzyme in saliva. Studies in humans found variation in enzymatic activity of sAA across populations that could be linked to the copy number of loci for salivary amylase (AMY1, which was seen as an adaptive response to the intake of dietary starch. In addition to diet dependent variation, differences in sAA activity have been related to social stress. In a previous study, we found evidence for stress-induced variation in sAA activity in the bonobos, a hominoid primate that is closely related to humans. In this study, we explored patterns of variation in sAA activity in bonobos and three other hominoid primates, chimpanzee, gorilla, and orangutan to (a examine if within-species differences in sAA activity found in bonobos are characteristic for hominoids and (b assess the extent of variation in sAA activity between different species. The results revealed species-differences in sAA activity with gorillas and orangutans having higher basal sAA activity when compared to Pan. To assess the impact of stress, sAA values were related to cortisol levels measured in the same saliva samples. Gorillas and orangutans had low salivary cortisol concentrations and the highest cortisol concentration was found in samples from male bonobos, the group that also showed the highest sAA activity. Considering published information, the differences in sAA activity correspond with differences in AMY1 copy numbers and match with general features of natural diet. Studies on sAA activity have the potential to complement molecular studies and may contribute to research on feeding ecology and nutrition.

  8. Age differences of salivary alpha-amylase levels of basal and acute responses to citric acid stimulation between Chinese children and adults

    Directory of Open Access Journals (Sweden)

    Zemin eYang

    2015-11-01

    Full Text Available It remains unclear how salivary alpha-amylase (sAA levels respond to mechanical stimuli in different age groups. In addition, the role played by the sAA gene (AMY1 copy number and protein expression (glycosylated and non-glycosylated in sAA activity has also been rarely reported. In this study, we analyzed saliva samples collected before and after citric acid stimulation from 47 child and 47 adult Chinese subjects. We observed that adults had higher sAA activity and sAA glycosylated levels (glycosylated sAA amount/total sAA amount in basal and stimulated saliva when compared with children, while no differences were found in total or glycosylated sAA amount between them. Interestingly, adults showed attenuated sAA activity levels increase over those of children after stimulation. Correlation analysis showed that total sAA amount, glycosylated sAA amount, and AMY1 copy number×total sAA amount were all positively correlated with sAA activity before and after stimulation in both groups. Interestingly, correlation r between sAA levels (glycosylated sAA amount and total sAA amount and sAA activity decreased after stimulation in children, while adults showed an increase in correlation r. In addition, the correlation r between AMY1 copy number×total sAA amount and sAA activity was higher than that between AMY1 copy number, total sAA amount and sAA activity, respectively. Taken together, our results suggest that total sAA amount, glycosylated sAA amount, and the positive interaction between AMY1 copy number and total sAA amount are crucial in influencing sAA activity before and after stimulation in children and adults.

  9. Display of alpha-amylase on the surface of Corynebacterium glutamicum cells by using NCgl1221 as the anchoring protein, and production of glutamate from starch.

    Science.gov (United States)

    Yao, Wenjuan; Chu, Chunli; Deng, Xiaozhao; Zhang, Yun; Liu, Miao; Zheng, Pu; Sun, Zhihao

    2009-10-01

    We developed a new cell surface display system in Corynebacterium glutamicum based on the C-terminally truncated NCgl1221 anchor protein to increase L-glutamate production from starch directly. The C-terminally truncated NCgl1221 protein is a mutant NCgl1221 and leads to the constitutive export of L-glutamate. The N terminus of alpha-amylase (AmyA) was fused to truncated NCgl1221, and the resulting fusion protein was expressed on the cell surface by IPTG induction. Localization of the fusion protein was confirmed by immunofluorescence microscopy and flow cytometric analysis. The results of L-glutamate fermentation showed that the soluble starch was utilized to grow and produce L-glutamate by the recombinant strain displaying AmyA. The amount of soluble starch was reduced from 30.0 +/- 2.8 to 4.5 +/- 0.7 g/l under non-inducing condition and from 50.0 +/- 2.4 to 12.5 +/- 1.1 g/l under biotin limitation in 36 h. The glutamate concentration in the medium was transiently increased in 14 h under no induction, while under biotin-limiting condition, glutamate production was continuously elevated during fermentation. The amount of glutamate reached 19.3 +/- 2.1 g/l after 26 h of fermentation with biotin limitation, which was greater than that produced by the strain using PgsA, one of the poly-gamma-glutamate synthetase complexes, as the anchor protein under the same condition. Therefore, the truncated NCgl1221 anchor protein has more advantages than the PgsA anchor protein in glutamate fermentation because truncated NCgl1221 leads to the constitutive export of L-glutamate without any treatments.

  10. Fortified complementary foods with or without alpha-amylase treatment increase hemoglobin but do not reduce breast milk intake of 9-mo-old Zambian infants.

    Science.gov (United States)

    Owino, Victor O; Kasonka, Lackson M; Sinkala, Moses M; Wells, Jonathan K; Eaton, Simon; Darch, Tegan; Coward, Andrew; Tomkins, Andrew M; Filteau, Suzanne M

    2007-10-01

    Malnutrition in late infancy in developing countries may result from poor-quality complementary foods that displace breast milk. The objective of the study was to assess the effects of fortified complementary blends of different energy densities on growth, hemoglobin concentrations, and breast milk intake of 9-mo-old Zambian infants. Infants were randomly assigned at 6 mo of age to receive for 3 mo a fortified blend of maize, beans, bambaranuts, and groundnuts [Chilenje Baby Mix (CBM); energy density: 68 kcal/100 g; n = 37] or a similar blend with alpha-amylase (CBMA; energy density: 106 kcal/100 g; n = 44). Cross-sectional data were obtained at 9 mo for a control group of infants (n = 69) not given the diets. Breast milk intake was measured by using the dose-to-the-mother deuterium dilution technique. No differences in weight or length z scores, all of which were within normal ranges, were seen between groups at 9 mo. Percentage fat mass was significantly (P = 0.01) greater in the infants in both the CBM (23.2 +/- 2.7%) and CBMA (23.4 +/- 2.5%) groups than in the control group (21.6 +/- 2.6%). Hemoglobin concentrations were significantly (P = 0.03) greater in both intervention groups (CBM group: 104 +/- 12 g/L: CBMA group: 103 +/- 12 g/L) than in the control group (98 +/- 14 g/L). Breast milk intake was not significantly (P = 0.87) different between groups (CBM group: 614 +/- 271 g/d; CBMA group: 635 +/- 193 g/d; control group: 653 +/- 221 g/d). The study foods improved hemoglobin concentrations without reducing breast milk intake and may be used to improve the nutritional status of infants in developing countries.

  11. 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.

  12. Effect of chronic training on heart rate variability, salivary IgA and salivary alpha-amylase in elite swimmers with a disability.

    Directory of Open Access Journals (Sweden)

    Rohan Edmonds

    Full Text Available The purpose of this study was to a determine the heart rate variability (HRV and saliva markers of immunity (salivary immunoglobulin A; sIgA and stress (salivary alpha-amylase; sAA responses to chronic training in elite swimmers with a disability; and b identify the relationships between HRV, sIgA, sAA and training volume. Eight members of a high performance Paralympic swimming program were monitored for their weekly resting HRV, sIgA and sAA levels in the 14 weeks leading up to a major international competition. The 14 week training program included aerobic, anaerobic, power and speed, and taper training phases, while also incorporating two swimming step tests and two swimming competitions. Specific time (root mean square of the successive differences; RMSSD and frequency (high frequency normalized units [HFnu] domain measures, along with non-linear indices (standard deviation of instantaneous RR variability; SD1 and short term fractal scaling exponent; α1 of HRV were used for all analyses with effects examined using magnitude-based inferences. Relationships between HRV and saliva markers were identified by Spearman rank rho (ρ correlation coefficients. Compared with week 1, SD1 was very likely lower (96/4/0, ES = -2.21, while sAA was very likely elevated (100/0/0, ES = 2.32 at the beginning of week 7 for all athletes. The training program did not alter HRV or saliva whereas competition did. There were also no apparent differences observed for HRV, sIgA and sAA between each of the training phases during the 14 week swimming program. Correlations were observed between sAA and SD1 (ρ = -0.212, p<0.05, along with sAA and mean HR (ρ = 0.309, p<0.05. These results show that high level national competition influences depresses HRV (SD1 and increases saliva biomarkers of stress (sAA. It appears that a well-managed and periodised swimming program can maintain these indices within normal baseline levels. The study also highlighted the

  13. Two secondary carbohydrate binding sites on the surface of barley alpha-amylase 1 have distinct functions and display synergy in hydrolysis of starch granules.

    Science.gov (United States)

    Nielsen, Morten M; Bozonnet, Sophie; Seo, Eun-Seong; Mótyán, János A; Andersen, Joakim M; Dilokpimol, Adiphol; Abou Hachem, Maher; Gyémánt, Gyöngyi; Naested, Henrik; Kandra, Lili; Sigurskjold, Bent W; Svensson, Birte

    2009-08-18

    Some polysaccharide processing enzymes possess secondary carbohydrate binding sites situated on the surface far from the active site. In barley alpha-amylase 1 (AMY1), two such sites, SBS1 and SBS2, are found on the catalytic (beta/alpha)(8)-barrel and the noncatalytic C-terminal domain, respectively. Site-directed mutagenesis of Trp(278) and Trp(279), stacking onto adjacent ligand glucosyl residues at SBS1, and of Tyr(380) and His(395), making numerous ligand contacts at SBS2, suggested that SBS1 and SBS2 act synergistically in degradation of starch granules. While SBS1 makes the major contribution to binding and hydrolysis of starch granules, SBS2 exhibits a higher affinity for the starch mimic beta-cyclodextrin. Compared to that of wild-type AMY1, the K(d) of starch granule binding by the SBS1 W278A, W279A, and W278A/W279A mutants thus increased 15-35 times; furthermore, the k(cat)/K(m) of W278A/W279A was 2%, whereas both affinity and activity for Y380A at SBS2 were 10% of the wild-type values. Dual site double and triple SBS1/SBS2 substitutions eliminated binding to starch granules, and the k(cat)/K(m) of W278A/W279A/Y380A AMY1 was only 0.4% of the wild-type value. Surface plasmon resonance analysis of mutants showed that beta-cyclodextrin binds to SBS2 and SBS1 with K(d,1) and K(d,2) values of 0.07 and 1.40 mM, respectively. A model that accounts for the observed synergy in starch hydrolysis, where SBS1 and SBS2 bind ordered and free alpha-glucan chains, respectively, thus targeting the enzyme to single alpha-glucan chains accessible for hydrolysis, is proposed. SBS1 and SBS2 also influence the kinetics of hydrolysis for amylose and maltooligosaccharides, the degree of multiple attack on amylose, and subsite binding energies.

  14. Salivary alpha amylase not chromogranin A reflects sympathetic activity: exercise responses in elite male wheelchair athletes with or without cervical spinal cord injury.

    Science.gov (United States)

    Leicht, Christof A; Paulson, Thomas A W; Goosey-Tolfrey, Victoria L; Bishop, Nicolette C

    2017-12-01

    Salivary alpha amylase (sAA) and chromogranin A (sCgA) have both been suggested as non-invasive markers for sympathetic nervous system (SNS) activity. A complete cervical spinal cord injury leading to tetraplegia is accompanied with sympathetic dysfunction; the aim of this study was to establish the exercise response of these markers in this in vivo model. Twenty-six elite male wheelchair athletes (C6-C7 tetraplegia: N = 8, T6-L1 paraplegia: N = 10 and non-spinal cord injured controls: N = 8) performed treadmill exercise to exhaustion. Saliva and blood samples were taken pre, post and 30 min post exercise and analysed for sAA, sCgA and plasma adrenaline concentration, respectively. In all three subgroups, sAA and sCgA were elevated post exercise (P < 0.05). Whilst sCgA was not different between subgroups, a group × time interaction for sAA explained the reduced post-exercise sAA activity in tetraplegia (162 ± 127 vs 313 ± 99 (paraplegia) and 328 ± 131 U mL -1 (controls), P = 0.005). The post-exercise increase in adrenaline was not apparent in tetraplegia (P = 0.74). A significant correlation was found between adrenaline and sAA (r = 0.60, P = 0.01), but not between adrenaline and sCgA (r = 0.06, P = 0.79). The blunted post-exercise rise in sAA and adrenaline in tetraplegia implies that both reflect SNS activity to some degree. It is questionable whether sCgA should be used as a marker for SNS activity, both due to the exercise response which is not different between the subgroups and its non-significant relationship with adrenaline.

  15. Effects of strategic early-morning caffeine gum administration on association between salivary alpha-amylase and neurobehavioural performance during 50 h of sleep deprivation.

    Science.gov (United States)

    Pajcin, Maja; White, Jason M; Banks, Siobhan; Dorrian, Jill; Paech, Gemma M; Grant, Crystal L; Johnson, Kayla; Tooley, Katie; Aidman, Eugene; Fidock, Justin; Kamimori, Gary H; Della Vedova, Chris B

    2018-02-02

    Self-assessment is the most common method for monitoring performance and safety in the workplace. However, discrepancies between subjective and objective measures have increased interest in physiological assessment of performance. In a double-blind placebo-controlled study, 23 healthy adults were randomly assigned to either a placebo (n = 11; 5 F, 6 M) or caffeine condition (n = 12; 4 F, 8 M) while undergoing 50 h (i.e. two days) of total sleep deprivation. In previous work, higher salivary alpha-amylase (sAA) levels were associated with improved psychomotor vigilance and simulated driving performance in the placebo condition. In this follow-up article, the effects of strategic caffeine administration on the previously reported diurnal profiles of sAA and performance, and the association between sAA and neurobehavioural performance were investigated. Participants were given a 10 h baseline sleep opportunity (monitored via standard polysomnography techniques) prior to undergoing sleep deprivation (total sleep time: placebo = 8.83 ± 0.48 h; caffeine = 9.01 ± 0.48 h). During sleep deprivation, caffeine gum (200 mg) was administered at 01:00 h, 03:00 h, 05:00 h, and 07:00 h to participants in the caffeine condition (n = 12). This strategic administration of caffeine gum (200 mg) has been shown to be effective at maintaining cognitive performance during extended wakefulness. Saliva samples were collected, and psychomotor vigilance and simulated driving performance assessed at three-hour intervals throughout wakefulness. Caffeine effects on diurnal variability were compared with previously reported findings in the placebo condition (n = 11). The impact of caffeine on the circadian profile of sAA coincided with changes in neurobehavioural performance. Higher sAA levels were associated with improved performance on the psychomotor vigilance test during the first 24 h of wakefulness in the caffeine condition

  16. Solid Culturing of Bacillus amyloliquefaciens for α-Amylase Production

    Directory of Open Access Journals (Sweden)

    Dhanya Gangadharan

    2006-01-01

    Full Text Available Fourteen different agroresidues were screened for alpha amylase production using Bacillus amyloliquefaciens ATCC 23842. Among them, wheat bran (WB and groundnut oil cake (GOC in mass ratio of 1:1 was proved as the best substrate source. Supplementation with 0.01 M KH2PO4 and 1 % soluble starch enhanced the enzyme yield considerably. Maximum enzyme recovery from the solid mass was obtained when extracted with 0.1 M acetate buffer, pH=5.0. Maximum enzyme titer expressed as units per mass of dry substrate obtained was 62 470 U/g after 72 hours of fermentation at 37 °C by using the above solid substrate mixture (5 g with the initial moisture of 85 % and inoculated with Bacillus amyloliquefaciens of 2·109 CFU/mL.

  17. Comportamiento de glicinina, beta-conglicinina y alfa-amilasa en semillas de soja deterioradas y no deterioradas Glycinin, beta-conglycinin and alpha-amylase behaviour in artificially deteriorated and not deteriorated soybean

    Directory of Open Access Journals (Sweden)

    Adriana Rita Salinas

    2002-08-01

    Full Text Available El objetivo del trabajo fue estudiar el comportamiento de glicinina y beta-conglicinina y la actividad de alfa-amilasa en semillas deterioradas y no deterioradas de 10 cultivares de soja [Glycine max (L. Merr.]. Las semillas se sometieron a dos tratamientos: deterioradas por envejecimiento acelerado y no deterioradas. Se determinó la presencia de las proteínas de reserva a partir de semillas con 0, 3 y 8 días de germinadas por electroforesis en geles de poliacrilamida (SDS-PAGE. La actividad de la alfa-amilasa se determinó bioquímicamente en semillas con 0, 3, 8 y 12 días de germinadas. No hubo diferencias en la presencia de bandas de glicinina y beta-conglicinina en semillas no deterioradas hasta los 8 días de germinadas. Las semillas deterioradas se comportaron en forma similar a las no deterioradas. La actividad de la alfa-amilasa aumentó en semillas germinadas hasta 8 días y disminuyó a los 12 días. En semillas deterioradas la actividad enzimática disminuyó con respecto a las no deterioradas. El deterioro artificial no afectó la presencia de glicinina y beta-conglicinina pero alteró la actividad de la alfa-amilasa hasta los 12 días de germinación. Los cultivares estudiados mostraron comportamiento diferencial frente a la actividad de esta enzima.The objective of this research was to study the behaviour of the storage proteins glycinin and beta-conglycinin and the alpha-amylase activity in artificially deteriorated and not deteriorated soybean [Glycine max (L. Merr.] seeds of 10 cultivars. The seeds were submitted to two treatments: deteriorated by accelerated aging and not deteriorated seeds. The presence of glycinin and beta-conglycinin were determined in seeds with 0, 3 and 8 days of germination using polyacrylamid gel electrophoresis (SDS-PAGE. The alpha-amylase activity was biochemically determined in seeds with 0, 3, 8 and 12 days of germination. There were no differences in the presence of glycinin and beta

  18. Effect of the medium composition on formation of amylase by Bacillus sp

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    Eliana de Oliveira. Santos

    2003-01-01

    Full Text Available Studies on the alpha -amylase synthesis was carried out with a moderately thermophilic, facultatively anaerobic Bacillus sp, isolated from soil samples. The cells were cultivated in a complex medium containing soluble starch or maltose as carbon source. The levels of the alpha -amylaseactivity detected in culture supernatants varied greatly with the type of carbon source used. Maltose, soluble starch and citrate stimulated alpha -amylaseformation. Addition of exogenous glucose repressed formation of alpha -amylase, demonstrating that a classical glucose effect was operative in this organism. The concentration of yeast extract was found to be important factor in the alpha -amylase synthesis bythe isolate.The activity of the enzyme increased between 2 and 5 g/L yeast extract concentration and then fell very rapidly beyond this point. The best concentration of peptone to alpha-amylase formation was found to be around 10g/L.Estudos sobre a síntese de alfa -amilase foram realizados com uma bactéria termofílica moderada e facultativa anaeróbica, isolada de amostras de solo. As células foram cultivadas em um meio complexo contendo amido solúvel ou maltose como fonte de carbono. Os níveis da atividade de alfa -amilase detectados no sobrenadante da cultura variaram grandemente com o tipo da fonte de carbono utilizada. Amido solúvel, maltose e citrato estimularam a formação de alfa -amilase. A adição de glicose as culturas reprimiu a formação da alfa -amilase, demonstrando que o clássico efeito glicose foi operativo neste organismo. A concentração de extrato de levedura foi um fator importante na formação de alfa -amilase pelo isolado. A atividade da enzima aumentou entre concentrações de 2 a 5 g/L e então caiu muito rapidamente em torno deste ponto. A melhor concentração de peptona para a formação da alfa -amilase foi em torno de 10 g/L.

  19. No relationship between baseline salivary alpha-amylase and State-Trait Anxiety Inventory Score in drug-naïve patients with short-illness-duration first episode major depressive disorder: An exploratory study.

    Science.gov (United States)

    Szarmach, Joanna; Cubała, Wiesław-Jerzy; Landowski, Jerzy; Chrzanowska, Anna

    2017-04-01

    Salivary α-amylase (sAA) activity alternations are observed in major depressive disorder (MDD) being associated with depression severity and its specific psychopathological dimensions with anxiety being attributed to distress. No data is available on sAA in MDD according to Hamilton Rating Scale for Depression (HAMD-17) and State-Trait Anxiety Inventory (STAI). The exploratory study examines whether and to what extent baseline sAA level is interrelated to the psychopathological features including severity of symptoms and specific psychopathological dimensions. The basal, non-stimulated sAA activity was studied in 20 non-late-life adult, treatment-naïve MDD patients with short-illness-duration and in 20 age- and sex-matched healthy controls along with psychometric assessments with Hamilton Rating Scale for Depression (HAMD-17) and Spielberger State-Trait Anxiety Inventory (STAI). Significantly lower ( p =0.011) sAA activity was observed in MDD as compared to controls. No significant correlations were observed between sAA activity and the total HAMD-17 score as well as with regard to the specific core depression, insomnia, anxiety and somatic HAM-D psychopathological dimensions. No significant correlations were also found between sAA and STAIX-1 and STAIX-2 scores. Low baseline sAA levels in MDD with no correlations between sAA and psychopathological features including severity of symptoms and specific psychopathological dimensions was found. Key words: Salivary alpha-amylase, major depressive disorder, Spielberger State-Trait Anxiety Inventory, Hamilton Rating Scale for Depression.

  20. An experimental study of the job demand-control model with measures of heart rate variability and salivary alpha-amylase: Evidence of increased stress responses to increased break autonomy.

    Science.gov (United States)

    O'Donnell, Emma; Landolt, Kathleen; Hazi, Agnes; Dragano, Nico; Wright, Bradley J

    2015-01-01

    We assessed in an experimental design whether the stress response towards a work task was moderated by the autonomy to choose a break during the assigned time to complete the task. This setting is defined in accordance with the theoretical framework of the job-demand-control (JDC) model of work related stress. The findings from naturalistic investigations of a stress-buffering effect of autonomy (or 'buffer hypothesis') are equivocal and the experimental evidence is limited, especially with relation to physiological indices of stress. Our objective was to investigate if increased autonomy in a particular domain (break time control) was related with adaptive physiology using objective physiological markers of stress; heart rate variability (HRV) and salivary alpha amylase (sAA). We used a within-subject design and the 60 female participants were randomly assigned to an autonomy (free timing of break) and standard conditions (fixed timing of break) of a word processing task in a simulated office environment in a random order. Participants reported increased perceptions of autonomy, no difference in demand and performed worse in the task in the break-time autonomy versus the standard condition. The results revealed support for the manipulation of increased autonomy, but in the opposing direction. Increased autonomy was related with dysregulated physiological reactivity, synonymous with typical increased stress responses. Potentially, our findings may indicate that autonomy is not necessary a resource but could become an additional stressor when it adds additional complexity while the amount of work (demands) remains unchanged. Further, our findings underscore the need to collect objective physiological evidence of stress to supplement self-reported information. Self-report biases may partially explain the inconsistent findings with the buffer hypothesis. Copyright © 2014 Elsevier Ltd. All rights reserved.

  1. Alpha-amylase inhibition kinetics by caulerpenyne

    OpenAIRE

    CENGIZ, S.; CAVAS, L.; YURDAKOC, K.

    2010-01-01

    Many algae have important secretions which are generally used for defensive purposes. These secretions take attentions of a lot of researchers who are wondering if these metabolites can be used for medical researches or not. Among these metabolites, caulerpenyne (CYN) which is the main metabolite of Caulerpa species, have had an important place in Caulerpa researches since the results related to its determined properties such as cytotoxic, antiviral, antiproliferative and apoptotic effects ha...

  2. Optimization of extraction parameters for recovery of a-amylase from the fermented bran of Bacillus circulans GRS313

    Directory of Open Access Journals (Sweden)

    Palit Soumen

    2001-01-01

    Full Text Available The extraction of alpha-amylase in the solid state fermentation of wheat bran by Bacillus circulans GRS313 was optimized. Among various solvents tested, maximum extraction was achieved when 2.5% (v/v glycerol was added. The optimum conditions for extraction were 2.5 hrs soaking time at 30° C under recirculation and agitated condition, which yielded 590 U/g bran of enzyme activity. Whereas under stationary soaking condition the activity of the enzyme was 439.58U/g bran. With repeated wash under the above optimum conditions showed that 600 U/g and 184.2 U/g of enzyme extracted in the 1st and 2nd washes respectively and only 36.84 U/g was recovered in the 3rd wash

  3. 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

  4. Alfa-amilase em frangos de corte: efeitos do balanço eletrolítico e do nível protéico da dieta Alpha-amylase in broiler chickens: effects of electrolytic balance and dietary protein level

    Directory of Open Access Journals (Sweden)

    Marcela Piedade Monteiro

    2006-06-01

    Full Text Available Um experimento foi conduzido com pintos de corte macho para o estudo dos efeitos dos níveis de 20 e 23% de PB combinados com 0, 50, 100, 150, 200, 250 mEq/kg de balanço dietético eletrolítico (BDE sobre a atividade da alfa-amilase pancreática de frangos de corte de 1 a 21 dias de idade. O delineamento utilizado foi o inteiramente casualizado. Dietas e água foram fornecidas ad libitum. Aos 1, 7, 14 e 21 dias, três aves de cada tratamento foram sacrificadas por deslocamento cervical para remoção do pâncreas, o qual foi removido, homogeneizado, congelado em nitrogênio líquido e liofilizado. Uma alíquota de cada amostra foi solubilizada em água deionizada e centrifugada a 7500 x g por 3 minutos a 4ºC, para determinação da atividade da alfa-amilase no sobrenadante. Aves alimentadas com 20% de PB apresentaram atividades específicas (U/mg de proteína superiores à daquelas que receberam 23%, exceto para os níveis de BDE de 100 a 200 mEq/kg, aos 14 dias. Observou-se tendência de aumento da atividade específica dos 7 aos 14 dias e redução dos 14 aos 21 dias. Para maior atividade específica de alfa-amilase, recomendam-se dietas com 20% de PB e 200 mEq/kg de BDE na fase pré-inicial e dietas com 20% de PB e 135 a 250 mEq/kg de BDE para frangos de corte dos 8 aos 21 dias de idade.An experiment was conducted with male broiler chicks to study the effects of 20 and 23% of crude protein (CP combined with 0, 50, 100, 150, 200, 250 mEq/kg of dietary electrolytic balance (DEB on the pancreatic alpha-amylase activity from 1 to 21 days. A completely randomized factorial design was used. Feed and water were supplied ad libitum. At days 1, 7, 14 and 21, three birds from each treatment were sacrificed by cervical dislocation, pancreas were removed, homogeneized and frozen in liquid nitrogen which were then freeze-dried. An aliquot of each sample was solubilized with deionized water and centrifuged at 7500 x g for three minutes at 4º

  5. Syntrophic co-culture of aerobic Bacillus and anaerobic Clostridium for bio-fuels and bio-hydrogen production

    Energy Technology Data Exchange (ETDEWEB)

    Chang, Jui-Jen; Ho, Cheng-Yu.; Chen, Wei-En; Huang, Chieh-Chen [Department of Life Sciences, National Chung Hsing University, Taichung (China); Chou, Chia-Hung; Lay, Jiunn-Jyi [Department of Science and Technology, National Kaohsiung First University, Kaohsiung (China)

    2008-10-15

    By using brewery yeast waste and microflora from rice straw compost, an anaerobic semi-solid bio-hydrogen-producing system has been established. For the purpose of industrialization, the major players of both aerobic and anaerobic bacterial strains in the system were isolated and their combination for an effective production of bio-hydrogen and other bio-fuels was examined in this study. The phylogenetic analysis found that four anaerobic isolates (Clostridium beijerinckii L9, Clostridium diolis Z2, Clostridium roseum Z5-1, and C. roseum W8) were highly related with each other and belongs to the cluster I clostridia family, the family that many of solvent-producing strains included. On the other hand, one of the aerobic isolates, the Bacillus thermoamylovorans strain I, shown multiple extracellular enzyme activities including lipase, protease, {alpha}-amylase, pectinase and cellulase, was suggested as a good partner for creating an anaerobic environment and pre-saccharification of substrate for those co-cultured solventogenic clostridial strain. Among these clostridial strains, though C. beijerinckii L9 do not show as many extracellular enzyme activities as Bacillus, but it performs the highest hydrogen-producing ability. The original microflora can be updated to a syntrophic bacterial co-culture system contended only with B. thermoamylovorans I and C. beijerinckii L9. The combination of aerobic Bacillus and anaerobic Clostridium may play the key role for developing the industrialized bio-fuels and bio-hydrogen-producing system from biomass. (author)

  6. Bacillus solimangrovi sp. nov., isolated from mangrove soil.

    Science.gov (United States)

    Lee, Geun-Hye; Rhee, Moon-Soo; Chang, Dong-Ho; Kwon, Kae Kyoung; Bae, Kyung Sook; Yang, Seong-Hyun; Kim, Byoung-Chan

    2014-05-01

    Two novel bacterial strains, GH2-4T and GH2-5, were isolated from mangrove soil near the seashore of Weno island in Chuuk state, Micronesia, and were characterized by a polyphasic approach. The two strains were strictly aerobic, Gram-staining-positive, motile, endospore-forming rods that were catalase- and oxidase-positive. Colonies were circular, convex, stringy and transparent yellowish (GH2-4T) or opaque whitish (GH2-5). The 16S rRNA gene sequences of the two isolates were identical. The most closely related strains in terms of 16S rRNA gene sequence similarity were Bacillus kochii WCC 4582T, B. horneckiae DSM 23495T, B. azotoformans LMG 9581T, B. cohnii DSM 6307T and B. halmapalus DSM 8723T (95.6, 95.4, 95.4, 95.2 and 95.2% similarity, respectively). The partial groEL sequence of strain GH2-4T was identical to that of strain GH2-5 and showed <85% similarity to those of the most closely related strains. The isolates grew at pH 5-12 (optimal growth at pH 9), at 10-40 °C (optimum 30-35 °C) and at 0-9% (w/v) NaCl (optimum 1-3% NaCl). The cell-wall peptidoglycan of strains GH2-4T and GH2-5 contained meso-diaminopimelic acid and cell-wall hydrolysates contained ribose as a major sugar. The DNA G+C content was 36 mol%, and DNA-DNA relatedness between the isolates and five related reference strains was 20-24%. Strain GH2-4T exhibited 81% DNA-DNA relatedness with strain GH2-5. The major cellular fatty acids of both strains were iso-C15:0, iso-C16:0, iso-C14:0 and anteiso-C15:0 and the predominant menaquinone was MK-7. On the basis of the evidence from this polyphasic study, strains GH2-4T and GH2-5 (=KCTC 33143=JCM 18995=DSM 27084) represent a novel species of the genus Bacillus, for which the name Bacillus solimangrovi sp. nov. is proposed; the type strain is GH2-4T (=KCTC 33142T=JCM 18994T=DSM 27083T).

  7. Comparative study on production of α-Amylase from Bacillus licheniformis strains

    Directory of Open Access Journals (Sweden)

    Dibu Divakaran

    2011-12-01

    Full Text Available Alpha amylase (α-1, 4-glucan-glucanhydrolase, EC 3.2.1.1, an extracellular enzyme, degrades α, 1-4 glucosidic linkages of starch and related substrates in an endo-fashion producing oligosaccharides including maltose, glucose and alpha limit dextrin (7. The present study deals with the production and comparative study of production of α-amylase from two strains of Bacillus licheniformis, MTCC 2617 and 2618, by using four different substrates, starch, rice, wheat and ragi powder as carbon source by submerged fermentation. The effect of varying pH and incubation temperature, activator, inhibitor, and substrate concentration was investigated on the activity of α-amylase produced by MTCC strain 2618. The results shows that the production of the α-amylase by the B.licheniformis strain MTCC 2618, using four different substrates were found to be maximum (Starch 3.64 IU/ml/minutes, Rice powder 2.93 IU/ml/minutes, Wheat powder 2.67 IU/ml/minutes, Ragi powder 2.36 IU/ml/minutes on comparing the enzyme production of two strains. It was also observed that the maximum production was found on the 3rd day (i.e. 72 hr and characterization of crude enzyme revealed that optimum activity was at pH 7 and 37ºC.

  8. Bacillus Coagulans

    Science.gov (United States)

    ... and, as a result, is often misclassified as lactic acid bacteria such as lactobacillus. In fact, some commercial products ... sporogenes or "spore-forming lactic acid bacterium." Unlike lactic acid bacteria such as lactobacillus or bifidobacteria, Bacillus coagulans forms ...

  9. Bacillus cucumis

    Science.gov (United States)

    Kämpfer, Peter; Busse, Hans-Jürgen; Glaeser, Stefanie P; Kloepper, Joseph W; Hu, Chia-Hui; McInroy, John A

    2016-02-01

    A facultative anaerobic, Gram-positive staining, endospore-forming bacterium, isolated from the rhizosphere of cucumber ( Cucumis sativus ), was taxonomically investigated. Based on 16S rRNA gene sequence similarity comparisons, strain AP-6 T clustered together with other species of the genus Bacillus and showed highest similarities with Bacillus drentensis LMG 21831 T (99.1 %), Bacillus vireti LMG 21834 T (98.7 %) and Bacillus soli LMG 21838 T (98.5 %). The 16S rRNA gene sequence similarity to the sequences of the type strains of other species of the genus Bacillus was 98.5 % or less. Chemotaxonomic features supported the grouping of the strain in the genus Bacillus ; for example, the major fatty acids were anteiso-C 15 : 0 , iso-C 15 : 0 and C 16 : 0 , the polar lipid profile contained the major components diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine and an unidentified glycolipid, the major quinone was menaquinone MK-7 and the major compound in the polyamine pattern was spermidine. Additionally, DNA-DNA hybridization with B. drentensis LMG 21831 T , B. vireti LMG 21834 T and B. soli LMG 21838 T resulted in relatedness values that were clearly below 70 %. Physiological and biochemical test results were also different from those of the most closely related species. As a consequence, AP-6 T represents a novel species of the genus Bacillus , for which the name Bacillus cucumis sp. nov. is proposed, with AP-6 T ( = CIP 110974 T  = CCM 8651 T ) as the type strain.

  10. Optimization of the industrial production of bacterial alpha amylase ...

    African Journals Online (AJOL)

    STORAGESEVER

    -mineral media. The time course of fermentation in a bioreactor revealed that the highest yield (about 8 x 104 U/ml within 60 h) by strain SCH was obtained by applying: 3.5% initial starch, 2% additional starch after 19 h, 3 vvm aeration and 300 ...

  11. Interaction of europium and curium with alpha-amylase

    Energy Technology Data Exchange (ETDEWEB)

    Barkleit, Astrid [Helmholtz-Zentrum Dresden-Rossendorf e.V., Dresden (Germany). Div. Chemistry of the F-Elements; Heller, Anne [Technische Univ. Dresden (Germany). Inst. for Zoology, Molecular Cell Physiology and Endocrinology; Helmholtz-Zentrum Dresden-Rossendorf e.V., Dresden (Germany). Div. Biogeochemistry

    2016-07-01

    Time-resolved laser-induced fluorescence spectroscopy (TRLFS) revealed that Eu(III) and Cm(III) form two dominant species with the protein α-amylase (Amy): one with the coordination of a single carboxylate group of the protein and the other with three coordinating carboxylate groups.

  12. Fundamental conditions required in extracting an alpha-amylase ...

    African Journals Online (AJOL)

    Crude extracts of Cadaba farinosa, Forsk plant are used for their starch-reducing power in most tropic countries to liquefy/sweeten starch gruels. The extraction process is a time consuming osmotic rehydrating process characterized by fluctuating extract yield and consequently, an unstable inherent starch-reducing power.

  13. 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

  14. Fundamental conditions required in extracting an alpha-amylase ...

    African Journals Online (AJOL)

    DUDE

    . G. Cadaba concentration. Cc w/v 0.14. 0.2. H. Temperature. T. °C 30. 45 enhance the starch-reducing power of extracts. Preliminary studies were carried out to identify these extraction parameters which enhanced enzyme activity; the most ...

  15. Magnetic alginate microparticles for purification of .alpha.-amylases

    Czech Academy of Sciences Publication Activity Database

    Šafaříková, Miroslava; Roy, I.; Gupta, M. N.; Šafařík, Ivo

    2003-01-01

    Roč. 105, - (2003), s. 255-260 ISSN 0168-1656 R&D Projects: GA MŠk OC 523.80; GA AV ČR IBS6087204 Institutional research plan: CEZ:AV0Z6087904 Keywords : alginate * ferrofluid * amalyses Subject RIV: CE - Biochemistry Impact factor: 2.543, year: 2003

  16. Salivary Alpha-Amylase Reactivity in Breast Cancer Survivors

    OpenAIRE

    Wan, Cynthia; Couture-Lalande, Marie-?ve; Narain, Tasha A.; Lebel, Sophie; Bielajew, Catherine

    2016-01-01

    The two main components of the stress system are the hypothalamic-pituitary-adrenal (HPA) and sympathetic-adrenal-medullary (SAM) axes. While cortisol has been commonly used as a biomarker of HPA functioning, much less attention has been paid to the role of the SAM in this context. Studies have shown that long-term breast cancer survivors display abnormal reactive cortisol patterns, suggesting a dysregulation of their HPA axis. To fully understand the integrity of the stress response in this ...

  17. Production, purification and characterization of thermostable α-amylase from soil isolate Bacillus sp. strain B-10

    Directory of Open Access Journals (Sweden)

    Ravindra Nath Singh

    2016-04-01

    Full Text Available A bacterial strain B-10 that produces α-amylase was isolated from compost and kitchen waste receiving agricultural soil. Based on microbiological and biochemical tests the isolate B-10 was identified as Bacillus sp. Alpha-amylase produced by this isolate was purified by (NH42SO4 precipitation and DEAE cellulose ion-exchange chromatography showing 15.91 and 48.21 fold purification, respectively. SDS-PAGE of the purified enzyme confirmed the purification and monomeric nature of the enzyme. The purified α-amylase showed maximum activity at pH 7 and temperature 50°C. The enzyme was significantly active in the temperature range of 30-60°C for the studied period of 2 h. During the incubation of purified enzyme at pH ranging from 5 to 10 for 24 h the maximum stability was observed at pH 7 followed by pH 8, whereas at extreme pH, the stability was very poor. Km and Vmax were found to be 1.4 mg/mL and 6.2 U/mL, respectively.

  18. Preparation of a Microbial Time-temperature Indicator by Using the Vegetative Form of Bacillus amyloliquefaciens for Monitoring the Quality of Chilled Food Products

    Directory of Open Access Journals (Sweden)

    Samaneh Shahrokh Esfahani

    2017-04-01

    Full Text Available Background and Objective: Time-temperature indicators are used in smart packaging, and described as intelligent tools attached to the label of food products to monitor their timetemperature history. Since the previous studies on microbial time-temperature indicators were only based on pH-dependent changes, and they were long-time response indicators, in the present work, a new microbial time-temperature indicator was designed by using the alpha amylase activity of Bacillus amyloliquefaciens vegetative cells.Material and Methods: The designed time-temperature indicator system consists of Bacillus amyloliquefaciens, specific substrate medium and iodine reagent. The relation of the timetemperatureindicator’ response to the growth and metabolic activity (starch consumption and production of reduced sugars of Bacillus amyloliquefaciens was studied. In addition, the temperature dependence of the time-temperature indicator was considered at 8 and 28˚C. Finally, in order to adjust time-temperature indicator endpoint, the effect of the inoculum level was investigated at 8ºC.Results and Conclusion: In the designed system, a color change of an iodine reagent to yellow progressively occurs due to the starch hydrolysis. The effect of the inoculum level showed the negative linear relationship between the levels of Bacillus amyloliquefaciens inoculated in the medium and the endpoints of the time-temperature indicators. The endpoints were adjusted to 156, 72 and 36 hours at the inoculum levels of 102, 104 and 106 CFU ml-1, respectively. The main advantages of the time-temperature indicator is low cost and application for monitoring the quality of chilled food products.Conflict of interest: The authors declare no conflict of interest.

  19. Bacillus thuringiensis

    Science.gov (United States)

    Hollensteiner, Jacqueline; Wemheuer, Franziska; Harting, Rebekka; Kolarzyk, Anna M; Diaz Valerio, Stefani M; Poehlein, Anja; Brzuszkiewicz, Elzbieta B; Nesemann, Kai; Braus-Stromeyer, Susanna A; Braus, Gerhard H; Daniel, Rolf; Liesegang, Heiko

    2016-01-01

    Verticillium wilt causes severe yield losses in a broad range of economically important crops worldwide. As many soil fumigants have a severe environmental impact, new biocontrol strategies are needed. Members of the genus Bacillus are known as plant growth-promoting bacteria (PGPB) as well as biocontrol agents of pests and diseases. In this study, we isolated 267 Bacillus strains from root-associated soil of field-grown tomato plants. We evaluated the antifungal potential of 20 phenotypically diverse strains according to their antagonistic activity against the two phytopathogenic fungi Verticillium dahliae and Verticillium longisporum . In addition, the 20 strains were sequenced and phylogenetically characterized by multi-locus sequence typing (MLST) resulting in 7 different Bacillus thuringiensis and 13 Bacillus weihenstephanensis strains. All B. thuringiensis isolates inhibited in vitro the tomato pathogen V. dahliae JR2, but had only low efficacy against the tomato-foreign pathogen V. longisporum 43. All B. weihenstephanensis isolates exhibited no fungicidal activity whereas three B. weihenstephanensis isolates showed antagonistic effects on both phytopathogens. These strains had a rhizoid colony morphology, which has not been described for B. weihenstephanensis strains previously. Genome analysis of all isolates revealed putative genes encoding fungicidal substances and resulted in identification of 304 secondary metabolite gene clusters including 101 non-ribosomal polypeptide synthetases and 203 ribosomal-synthesized and post-translationally modified peptides. All genomes encoded genes for the synthesis of the antifungal siderophore bacillibactin. In the genome of one B. thuringiensis strain, a gene cluster for zwittermicin A was detected. Isolates which either exhibited an inhibitory or an interfering effect on the growth of the phytopathogens carried one or two genes encoding putative mycolitic chitinases, which might contribute to antifungal activities

  20. Biosynthesis of titanium dioxide nanoparticles using Bacillus amyloliquefaciens culture and enhancement of its photocatalytic activity for the degradation of a sulfonated textile dye Reactive Red 31.

    Science.gov (United States)

    Khan, Razia; Fulekar, M H

    2016-08-01

    The present study aims at exploiting Bacillus amyloliquefaciens for the biosynthesis of titanium dioxide nanoparticles and also investigates role of bacterial enzymes in the biosynthesis of titanium dioxide nanoparticles. Bacterial synthesized as well as metal doped titanium dioxide nanoparticles were characterized by X-ray diffractometer (XRD), Fourier transform infrared spectroscopy (FTIR), Transmission electron microscopy (TEM), Energy dispersive X-ray spectroscopy (EDAX). Amylase activity (43.37IU) in culture supernatant evinced a potential involvement of extracellular enzyme in TiO2 nanoparticle biosynthesis. Crystallite size of bio-synthesized nanoparticles was found to be in the range of 15.23-87.6nm. FTIR spectroscopy and native-PAGE (Polyacrylamide Gel Electrophoresis) clearly indicated involvement of alpha amylase in biosynthesis of TiO2 nanoparticles and in their stabilization. TEM micrographs of the synthesized titanium dioxide nanoparticles revealed the formation of spherical nanoparticles with a size range of 22.11-97.28nm. Photocatalytic degradation of Reactive Red 31 (RR31) dye was carried out using bio-synthesized TiO2 nanoparticles under UV radiation. Photocatalytic activity of synthesized nanoparticles was enhanced by Ag, La, Zn and Pt doping. Platinum doped TiO2 showed highest potential (90.98%) in RR31 degradation as compared to undoped (75.83%). Copyright © 2016 Elsevier Inc. All rights reserved.

  1. Bacillus amyloliquefaciens

    Science.gov (United States)

    Qin, Yuxuan; Shang, Qingmao; Zhang, Ying; Li, Pinglan; Chai, Yunrong

    2017-01-01

    Vegetable plug seedling has become the most important way to produce vegetable seedlings in China. This seedling method can significantly improve the quality and yield of vegetables compared to conventional methods. In the process of plug seedling, chemical fertilizers or pesticides are often used to improve the yield of the seedlings albeit with increasing concerns. Meanwhile, little is known about the impact of beneficial bacteria on the rhizosphere microbiota and the growth conditions of vegetables during plug seedling. In this study, we applied a culture-independent next-generation sequencing-based approach and investigated the impact of a plant beneficial bacterium, Bacillus amyloliquefaciens L-S60, on the composition and dynamics of rhizosphere microbiota and the growth conditions of cucumbers during plug seedling. Our results showed that application of L-S60 significantly altered the structure of the bacterial community associated with the cucumber seedling; presence of beneficial rhizosphere species such as Bacillus, Rhodanobacter, Paenibacillus, Pseudomonas, Nonomuraea , and Agrobacterium was higher upon L-S60 treatment than in the control group. We also measured the impact of L-S60 application on the physiological properties of the cucumber seedlings as well as the availability of main mineral elements in the seedling at different time points during the plug seedling. Results from those measurements indicated that L-S60 application promoted growth conditions of cucumber seedlings and that more available mineral elements were detected in the cucumber seedlings from the L-S60 treated group than from the control group. The findings in this study provided evidence for the beneficial effects of plant growth-promoting rhizosphere bacteria on the bacterial community composition and growth conditions of the vegetables during plug seedling.

  2. Bacillus subtilis

    Science.gov (United States)

    Wang, Xiaoqing; Hu, Weiwei; Zhu, Liqi; Yang, Qian

    2017-04-28

    Intestinal epithelial cells are the targets for transmissible gastroenteritis (TGE) virus (TGEV) infection. It is urgent to develop a novel candidate against TGEV entry. Bacillus subtilis is a probiotic with excellent anti-microorganism properties and one of its secretions, surfactin, has been regarded as a versatile weapon for most plant pathogens, especially for the enveloped virus. We demonstrate for the first time that B. subtilis OKB105 and its surfactin can effectively inhibit one animal coronavirus, TGEV, entering the intestinal porcine epithelial cell line (IPEC-J2). Then, several different experiments were performed to seek the might mechanisms. The plaque assays showed that surfactant could reduce the plaque generation of TGEV in a dose-dependent manner. Meanwhile, after incubation with TGEV for 1.5 h, B. subtilis could attach TGEV particles to their surface so that the number of virus to bind to the host cells was declined. Furthermore, our data showed that the inhibition of B. subtilis was closely related to the competition with TGEV for the viral entry receptors, including epidermal growth factor receptor (EGFR) and aminopeptidase N (APN) protein. In addition, Western blotting and apoptosis analysis indicated that B. subtilis could enhance the resistance of IPEC-J2 cells by up-regulating the expression of toll-like receptor (TLR)-6 and reducing the percentage of apoptotic cells. Taken together, our results suggest that B. subtilis OKB105 and its surfactin can antagonize TGEV entry in vitro and may serve as promising new candidates for TGEV prevention. © 2017 The Author(s).

  3. Bacillus coagulans

    Science.gov (United States)

    Aulitto, Martina; Fusco, Salvatore; Bartolucci, Simonetta; Franzén, Carl Johan; Contursi, Patrizia

    2017-01-01

    The transition from a petroleum-based economy towards more sustainable bioprocesses for the production of fuels and chemicals (circular economy) is necessary to alleviate the impact of anthropic activities on the global ecosystem. Lignocellulosic biomass-derived sugars are suitable alternative feedstocks that can be fermented or biochemically converted to value-added products. An example is lactic acid, which is an essential chemical for the production of polylactic acid, a biodegradable bioplastic. However, lactic acid is still mainly produced by Lactobacillus species via fermentation of starch-containing materials, the use of which competes with the supply of food and feed. A thermophilic and cellulolytic lactic acid producer was isolated from bean processing waste and was identified as a new strain of Bacillus coagulans , named MA-13. This bacterium fermented lignocellulose-derived sugars to lactic acid at 55 °C and pH 5.5. Moreover, it was found to be a robust strain able to tolerate high concentrations of hydrolysate obtained from wheat straw pre-treated by acid-catalysed (pre-)hydrolysis and steam explosion, especially when cultivated in controlled bioreactor conditions. Indeed, unlike what was observed in microscale cultivations (complete growth inhibition at hydrolysate concentrations above 50%), B. coagulans MA-13 was able to grow and ferment in 95% hydrolysate-containing bioreactor fermentations. This bacterium was also found to secrete soluble thermophilic cellulases, which could be produced at low temperature (37 °C), still retaining an optimal operational activity at 50 °C. The above-mentioned features make B. coagulans MA-13 an appealing starting point for future development of a consolidated bioprocess for production of lactic acid from lignocellulosic biomass, after further strain development by genetic and evolutionary engineering. Its optimal temperature and pH of growth match with the operational conditions of fungal enzymes hitherto

  4. Toxicity of Tolyltriazole to Bacillus Microorganisms.

    Science.gov (United States)

    2000-03-01

    Bacillus coagulans Microbacterium lacticum Jupiter Bacillus thuringiensis Bacillus thuringiensis Bacillus cereus Bacillus Bacillus thuringiensis...TOXICITY OF TOLYLTRIAZOLE TO BACILLUS MICROORGANISMS THESIS Christopher J. Leonard, First Lieutenant, USAF AFIT/GEE/ENV/OOM-12 Approved for...AND DATES COVERED Master’s Thesis 4. TITLE AND SUBTITLE TOXICITY OF TOLYLTRIAZOLE TO BACILLUS MICROORGANISMS 6. AUTHOR(S) Christopher J

  5. Noncontiguous finished genome sequences and description of Bacillus massiliglaciei, Bacillus mediterraneensis, Bacillus massilinigeriensis, Bacillus phocaeensis and Bacillus tuaregi, five new species identified by culturomics

    OpenAIRE

    Cadoret, F.; Alou, M.T.; Afouda, P.; Traore, I.S.; Br?chard, L.; Michelle, C.; Di Pinto, F.; Andrieu, C.; Delerce, J.; Levasseur, A.; Fournier, P.-E.; Raoult, D.

    2017-01-01

    Microbial culturomics, which investigates microbial diversity by combining diversified culture conditions, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and 16S rDNA identification, allowed to identify five new species within the Bacillus genus. Bacillus massiliglaciei strain Marseille-P2600T, Bacillus mediterraneensis strain Marseille-P2384T, Bacillus massilinigeriensis strain Marseille-P2366T, Bacillus tuaregi strain Marseille-P2489T and Bacillus phocaeensis s...

  6. Siderophores of Bacillus anthracis, Bacillus cereus, and Bacillus thuringiensis.

    Science.gov (United States)

    Wilson, Melissa K; Abergel, Rebecca J; Raymond, Kenneth N; Arceneaux, Jean E L; Byers, B Rowe

    2006-09-15

    Three Bacillus anthracis Sterne strains (USAMRIID, 7702, and 34F2) and Bacillus cereus ATCC 14579 excrete two catecholate siderophores, petrobactin (which contains 3,4-dihydroxybenzoyl moieties) and bacillibactin (which contains 2,3-dihydroxybenzoyl moieties). However, the insecticidal organism Bacillus thuringiensis ATCC 33679 makes only bacillibactin. Analyses of siderophore production by previously isolated [Cendrowski et al., Mol. Microbiol. 52 (2004) 407-417] B. anthracis mutant strains revealed that the B. anthracis bacACEBF operon codes for bacillibactin production and the asbAB gene region is required for petrobactin assembly. The two catecholate moieties also were synthesized by separate routes. PCR amplification identified both asbA and asbB genes in the petrobactin producing strains whereas B. thuringiensis ATCC 33679 retained only asbA. Petrobactin synthesis is not limited to the cluster of B. anthracis strains within the B. cereus sensu lato group (in which B. cereus, B. anthracis, and B. thuringiensis are classified), although petrobactin might be prevalent in strains with pathogenic potential for vertebrates.

  7. Phosphorescence In Bacillus Spores

    National Research Council Canada - National Science Library

    Reinisch, Lou; Swartz, Barry A; Bronk, Burt V

    2003-01-01

    .... Our present work attempts to build on this approach for environmental applications. We have measured a change in the fluorescence spectra of suspensions of Bacillus bacteria between the vegetative bacteria and their spores at room temperature...

  8. Properties of an amylase from thermophilic Bacillus SP. Propriedades de uma amilase de um termofílico Bacillus sp

    Directory of Open Access Journals (Sweden)

    Raquel Vieira de Carvalho

    2008-03-01

    Full Text Available alpha-Amylase production by thermophilic Bacillus sp strain SMIA-2 cultivated in liquid cultures containing soluble starch as a carbon source and supplemented with 0.05% whey protein and 0.2% peptone reached a maximum activity at 32 h, with levels of 37 U/mL. Studies on the amylase characterization revealed that the optimum temperature of this enzyme was 90ºC. The enzyme was stable for 1 h at temperatures ranging from 40-50ºC while at 90ºC, 66% of its maximum activity was lost. However, in the presence of 5 mM CaCl2, the enzyme was stable at 90ºC for 30 min and retained about 58% residual activity after 1 h. The optimum pH of the enzyme was found to be 8.5. After incubation of enzyme for 2 h at pH 9.5 and 11.0 was observed a decrease of about 6.3% and 16.5% of its original activity. At pH 6.0 the enzyme lost about 36% of its original activity. The enzyme was strongly inhibited by Co2+, Cu2+ and Ba2+, but less affected by Mg2+, Na+ and K+. In the presence of 2.0 M NaCl, 63% of amylase activity was retained after 2 h incubation at 45ºC. The amylase exhibited more than 70% activity when incubated for 1 h at 50ºC with sodium dodecyl sulphate. However, very little residual activity was obtained with sodium hypochlorite and with hydrogen peroxide the enzyme was completely inhibited. The compatibility of Bacillus sp SMIA-2 amylase with certain commercial detergents was shown to be good as the enzyme retained 86%, 85% and 75% of its activity after 20 min incubation at 50ºC in the presence of the detergent brands Omo®, Campeiro® and Tide®, respectively.A produção de alfa-amilase por um termofilico, Bacillus sp SMIA-2, cultivado em meio líquido contendo amido solúvel como fonte de carbono, alcançou uma atividade máxima de 37 U/mL em 32 horas. Estudos sobre a caracterização da amilase revelaram que a temperatura ótima desta enzima foi 90ºC. A enzima foi estável por 1 hora a temperaturas de 40 e 50ºC enquanto a 90ºC, 66% da atividade

  9. Bacillus odysseyi isolate

    Science.gov (United States)

    Venkateswaran, Kasthuri (Inventor); La Duc, Myron Thomas (Inventor)

    2007-01-01

    The present invention relates to discovery and isolation of a biologically pure culture of a Bacillus odysseyi isolate with high adherence and sterilization resistant properties. B. odysseyi is a round spore forming Bacillus species that produces an exosporium. This novel species has been characterized on the basis of phenotypic traits, 16S rDNA sequence analysis and DNA-DNA hybridization. According to the results of these analyses, this strain belongs to the genus Bacillus and the type strain is 34hs-1.sup.T (=ATCC PTA-4993.sup.T=NRRL B-30641.sup.T=NBRC 100172.sup.T). The GenBank accession number for the 16S rDNA sequence of strain 34hs-1.sup.T is AF526913.

  10. Bacillus velezensis is not a later heterotypic synonym of Bacillus amyloliquefaciens; Bacillus methylotrophicus, Bacillus amyloliquefaciens subsp plantarum and ‘Bacillus oryzicola’ are later heterotypic synonyms of Bacillus

    Science.gov (United States)

    The rhizosphere isolated bacteria belonging to the Bacillus amyloliquefaciens subsp. plantarum and Bacillus methylotrophicus clades are an important group of strains that are used as plant growth promoters and antagonists of plant pathogens. These properties have made these strains the focus of comm...

  11. Characterization of Bacillus cereus

    NARCIS (Netherlands)

    Wijnands LM; Dufrenne JB; Leusden FM; MGB

    2002-01-01

    Bacillus cereus is a ubiquitary microorganism that may cause food borne disease. Pathogenicity, however, depends on various characteristics such as the ability to form (entero)-toxin(s) that can not be detected by microbiological methods. Further characterization of pathogenic properties is not only

  12. Essential Bacillus subtilis genes

    NARCIS (Netherlands)

    Kobayashi, K.; Ehrlich, S.D.; Albertini, A.; Amati, G.; Andersen, K.K.; Arnaud, M.; Asai, K.; Ashikaga, S.; Aymerich, S.; Bessieres, P.; Boland, F.; Brignell, S.C.; Bron, S; Bunai, K.; Chapuis, J; Christiansen, L.C.; Danchin, A.; Debarbouille, M.; Dervyn, E.; Deuerling, E.; Devine, K.; Devine, S.K.; Dreesen, O.; Errington, J.; Fillinger, S.; Foster, S.J.; Fujita, Y.; Galizzi, A.; Gardan, R.; Eschevins, C.; Fukushima, T.; Haga, K.; Harwood, C.R; Hecker, M.; Hosoya, D.; Hullo, M.F.; Kakeshita, H.; Karamata, D.; Kasahara, Y.; Kawamura, F.; Koga, K.; Koski, P.; Kuwana, R.; Imamura, D.; Ishimaru, M.; Ishikawa, S.; Ishio, I.; Le Coq, D.; Masson, A.; Mauel, C.; Meima, Roelf; Mellado, R.P.; Moir, A.; Moriya, S.; Nagakawa, E.; Nanamiya, H.; Nakai, S.; Nygaard, P.; Ogura, M.; Ohanan, T.; O'Reilly, M.; O'Rourke, M.; Pragai, Z.; Pooley, H.M.; Rapoport, G.; Rawlins, J.P.; Rivas, L.A.; Rivolta, C.; Sadaie, A.; Sadaie, Y.; Sarvas, M; Sato, T.; Saxild, H.H.; Scanlan, E.; Schumann, W; Seegers, J.F. M. L.; Sekiguchi, J.; Sekowska, A.; Seror, S.J.; Simon, M.; Stragier, P.; Studer, R.; Takamatsu, H.; Tanaka, T.; Takeuchi, M.; Thomaides, H.B.; Vagner, V.; van Dijl, J.M.; Watabe, K.; Wipat, A; Yamamoto, H.; Yamamoto, M.; Yamamoto, Y.; Yamane, K.; Yata, K.; Yoshida, K.; Yoshikawa, H.; Zuber, U.; Ogasawara, N.; Ishio, [No Value

    2003-01-01

    To estimate the minimal gene set required to sustain bacterial life in nutritious conditions, we carried out a systematic inactivation of Bacillus subtilis genes. Among approximate to4,100 genes of the organism, only 192 were shown to be indispensable by this or previous work. Another 79 genes were

  13. Draft Genome Sequences of Three Alkaliphilic Bacillus Strains, Bacillus wakoensis JCM 9140T, Bacillus akibai JCM 9157T, and Bacillus hemicellulosilyticus JCM 9152T

    OpenAIRE

    Yuki, Masahiro; Oshima, Kenshiro; Suda, Wataru; Oshida, Yumi; Kitamura, Keiko; Iida, Toshiya; Hattori, Masahira; Ohkuma, Moriya

    2014-01-01

    Here, we report the draft genome sequences of the type strains of three cellulolytic or hemicellulolytic alkaliphilic Bacillus species: Bacillus wakoensis, Bacillus akibai, and Bacillus hemicellulosilyticus. The genome information for these three strains will be useful for studies of alkaliphilic Bacillus species, their evolution, and biotechnological applications for their enzymes.

  14. Taxonomy Icon Data: Bacillus subtilis [Taxonomy Icon

    Lifescience Database Archive (English)

    Full Text Available g Bacillus_subtilis_S.png Bacillus_subtilis_NS.png http://biosciencedbc.jp/taxonomy_icon/icon.cgi?i=Bacillus...+subtilis&t=L http://biosciencedbc.jp/taxonomy_icon/icon.cgi?i=Bacillus+subtilis&t=NL http://biosciencedbc.jp/taxonom...y_icon/icon.cgi?i=Bacillus+subtilis&t=S http://biosciencedbc.jp/taxonomy..._icon/icon.cgi?i=Bacillus+subtilis&t=NS http://togodb.biosciencedbc.jp/togodb/view/taxonomy_icon_comment_en?species_id=214 ...

  15. Microbial genotyping and differentiating between Bacillus mojavensis and Bacillus subtilis

    Science.gov (United States)

    Bacillus mojavensis, a specie recently distinguished from its previous Bacillus subtilis classification, was discovered in corn kernels and later determined to possess endophytic character. The bacterium was also determined to have biocontrol potential due to its growth inhibition of the maize mycot...

  16. 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.

  17. 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.

  18. Salivary alpha-amylase, heart rate and heart-rate variability in ...

    African Journals Online (AJOL)

    African Journal for Physical Activity and Health Sciences ... A competitive stress game model based on essential volleyball techniques was designed. ... HRavg and HRmax increased (p<0.001) during warm-up by 33% and by12.9% and during the experimental game by 21.2% and by 24% compared to baseline values ...

  19. Classroom Emotional Support Predicts Differences in Preschool Children's Cortisol and Alpha-Amylase Levels

    Science.gov (United States)

    Hatfield, Bridget E.; Hestenes, Linda L.; Kintner-Duffy, Victoria L.; O'Brien, Marion

    2013-01-01

    Accumulating evidence suggests children enrolled in full-time child care often display afternoon elevations of the hormone cortisol, which is an indicator of stress. Recent advances in immunoassays allow for measurement of activity in the hypothalamic-pituitary-adrenal axis and the autonomic sympathetic nervous system from saliva, and measurement…

  20. Sex Differences in Salivary Cortisol, Alpha-Amylase, and Psychological Functioning Following Hurricane Katrina

    Science.gov (United States)

    Vigil, Jacob M.; Geary, David C.; Granger, Douglas A.; Flinn, Mark V.

    2010-01-01

    The study examines group and individual differences in psychological functioning and hypothalamic-pituitary-adrenal and sympathetic nervous system (SNS) activity among adolescents displaced by Hurricane Katrina and living in a U.S. government relocation camp (n = 62, ages 12-19 years) 2 months postdisaster. Levels of salivary cortisol, salivary…

  1. 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.

  2. 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

    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......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...

  3. Interactions of barley alpha-amylase isozymes with Ca2+, substrates and proteinaceous inhibitors

    DEFF Research Database (Denmark)

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

    2006-01-01

    has revealed that AMY1 has ten functional subsites which can be modified by means protein engineering to modulate the substrate specificity. Other mutational analyses show that surface carbohydrate binding sites are critical for interaction with polysaccharides. The conserved Tyr380 in the newly...... discovered 'sugar tongs' site in domain C of AMY1 is thus critical for binding to starch granules. Furthermore, mutations of binding sites mostly reduced the degree of multiple attack in amylose hydrolysis. AMY1 has higher substrate affinity than AMY2, but isozyme chimeras with AMY2 domain C and other...

  4. The starch-bound alpha-amylase/trypsin-inhibitors in Avena.

    Science.gov (United States)

    Gazza, Laura; Gazzelloni, Gloria; Taddei, Federica; Latini, Arianna; Muccilli, Vera; Alfieri, Michela; Conti, Salvatore; Redaelli, Rita; Pogna, Norberto E

    2016-12-01

    Oat kernels exhibit an extra-soft texture, a trait recently demonstrated to be largely modulated by starch-bound tryptophan-rich 2S proteins, the vromindolines. In this study, fractionation by two-dimensional electrophoresis of starch-bound proteins in 25 oat (Avena sativa) cultivars and 11 diploid or tetraploid Avena species revealed novel 2S proteins called Avena α-amylase/trypsin-inhibitors (AATI) because of their sequence similarity with wheat α-amylase/trypsin inhibitors. Thirty-seven AATI polypeptides, about 14 kDa in size, were split into three families named AATI-1, AATI-2, and AATI-3 with different primary structures and isoelectric points. AATI-1 and AATI-2 proteins showed 55.5-60.0 % sequence similarity with wheat α-amylase inhibitors CM1, CM2, and CM16, which have been found to cause innate immunity responses in celiac disease and non-celiac gluten sensitivity. Diploid A-genome and tetraploid AC-genome oat species possess three and five genes encoding for the AATI proteins, respectively, whereas hexaploid A. sativa exhibits 12 genes dispersed over the A-, C-, and D-genomes. Some AATI proteins expressed in hexaploid oats were assigned to the A-genome based on similarity to their counterparts in diploid species, contributing to further clarify the genetic origin of hexaploid oats. Moreover, AATI may interact with starch-bound vromindolines in determining the extra-soft texture of oat kernels and, due to their balanced amino acid compositions, may contribute to the biological value of oat proteins in a positive manner.

  5. 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.

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

    Directory of Open Access Journals (Sweden)

    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.

  7. Salivary alpha-amylase : a measure associated with satiety and subsequent food intake in humans

    NARCIS (Netherlands)

    Harthoorn, L.F.

    2008-01-01

    Food intake regulation in humans involves various central and peripheral mechanisms. In this study salivary -amylase was examined for functioning as a measure of satiety and food intake. In a 1.25-h session, 32 fasted subjects were given a preload of starch-based custard (849 kJ) followed by ad

  8. Brain alpha-amylase - a novel energy regulator important in Alzheimer disease?

    NARCIS (Netherlands)

    Byman, Elin; Schultz, Nina; Huitinga, I.; Fex, Malin; Wennström, Malin

    2018-01-01

    Reduced glucose metabolism and formation of polyglucosan bodies (PGB) are, beside amyloid beta plaques and neurofibrillary tangles, well-known pathological findings associated with Alzheimer's disease (AD). Since both glucose availability and PGB are regulated by enzymatic degradation of glycogen,

  9. Inhibitory specificity and insecticidal selectivity of alpha-amylase inhibitor from Phaseolus vulgaris

    Czech Academy of Sciences Publication Activity Database

    Kluh, Ivan; Horn, Martin; Hýblová, Jana; Hubert, J.; Dolečková, Lucie; Voburka, Zdeněk; Kudlíková, I.; Kocourek, F.; Mareš, Michael

    2005-01-01

    Roč. 66, - (2005), 31-39 ISSN 0031-9422 R&D Projects: GA MŠk(CZ) OC D16.001 Institutional research plan: CEZ:AV0Z4055905 Keywords : enzyme inhibition * digestive enzyme Subject RIV: FD - Oncology ; Hematology Impact factor: 2.780, year: 2005

  10. De novo design of alpha-amylase inhibitor: A small linear mimetic of macromolecular proteinaceous ligands

    Czech Academy of Sciences Publication Activity Database

    Marešová, Lucie; Pavlík, Manfred; Horn, Martin; Mareš, Michael

    2005-01-01

    Roč. 12, č. 12 (2005), 1349-1357 ISSN 1074-5521 R&D Projects: GA ČR(CZ) GP203/02/P081; GA MŠk(CZ) OC D16.001 Institutional research plan: CEZ:AV0Z40550506 Keywords : amylase * peptide inhibitor * combinatorial chemistry Subject RIV: CE - Biochemistry Impact factor: 6.138, year: 2005

  11. 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...

  12. 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.

  13. Amylosucrase, a glucan-synthesizing enzyme from the alpha-amylase family

    DEFF Research Database (Denmark)

    Skov, L K; Mirza, Osman Asghar; Henriksen, A

    2001-01-01

    of amylosucrase is at the bottom of a pocket at the molecular surface. A substrate binding site resembling the amylase 2 subsite is not found in amylosucrase. The site is blocked by a salt bridge between residues in the second and eight loops of the (beta/alpha)(8)-barrel. The result is an exo-acting enzyme. Loop...... 7 in the amylosucrase barrel is prolonged compared with the loop structure found in other hydrolases, and this insertion (forming domain B') is suggested to be important for the polymer synthase activity of the enzyme. The topology of the B'-domain creates an active site entrance with several...

  14. 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

  15. 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

  16. Influence of carbon source on alpha-amylase production by Aspergillus oryzae

    DEFF Research Database (Denmark)

    Carlsen, Morten; Nielsen, Jens

    2001-01-01

    The influence of the carbon source on a-amylase production by Aspergillus oryzae was quantified in carbon-limited chemostat cultures. The following carbon sources were investigated: maltose, maltodextrin (different chain lengths), glucose, fructose, galactose, sucrose, glycerol, mannitol and acet...

  17. The use of starch azure for measurement of alpha-amylase activity.

    Science.gov (United States)

    Lehoczki, Gábor; Kandra, Lili; Gyémánt, Gyöngyi

    2018-03-01

    Despite being widely used, there is no standard protocol for α-amylase activity measurement with starch azure substrate. Boiling pre-treatment of starch azure suspension increased the reaction rate of hydrolysis catalysed by human salivary α-amylase (HSA) or porcine pancreatic α-amylase (PPA) and the sensitivity of spectrophotometric activity measurement has been improved. Kinetic constants, K M , and v max , obtained from parallel isothermal titration calorimetric (ITC) measurements on natural and starch azure revealed, that the blue starch derivative does not differ significantly from its natural counterpart from kinetic point of view. Interestingly, substrate inhibition was observed in starch azure hydrolysis characterised by dissociation constants 49 mg/mL and 16.4 mg/mL for HSA and PPA, respectively. In this work a new protocol has been suggested for α-amylase activity measurement using boiled insoluble starch azure as substrate at 5 mg/mL concentration. Copyright © 2017 Elsevier Ltd. All rights reserved.

  18. Controlled shielding and deshielding of gene delivery polyplexes using hydroxyethyl starch (HES) and alpha-amylase.

    Science.gov (United States)

    Noga, Matthäus; Edinger, Daniel; Rödl, Wolfgang; Wagner, Ernst; Winter, Gerhard; Besheer, Ahmed

    2012-04-10

    The non-viral delivery of nucleic acids faces many extracellular and intracellular hurdles on the way from injection site to the site of action. Among these, aggregation in the blood stream and rapid elimination by the mononuclear phagocytic system (MPS) represent strong obstacles towards successful development of these promising therapeutic modalities. Even the state-of-the-art solutions using PEGylation show low transfection efficiency due to limited uptake and hindered endosomal escape. Engineering the carriers with sheddable coats reduces aggregation and phagocytosis due to the effective shielding, while the controlled deshielding at the desired site of action enhances the uptake and intracellular release. This work reports for the first time the use of hydroxyethyl starch (HES) for the controlled shielding/deshielding of polyplexes. HES, with different molar masses, was grafted to polyethylenimine (PEI) and characterized using (1)H NMR, colorimetric copper-assay, and SEC. HES-PEI conjugates were used to generate polyplexes with the luciferase-expressing plasmid DNA pCMVluc, and were characterized by DLS and zeta potential measurements. Deshielding was tested in vitro by zeta potential measurements and, erythrocyte aggregation assay upon addition of α-amylase (AA) to the HES-decorated particles. The addition of AA led to gradual increase in the zeta potential of the nanoparticles over 0.5 to 1h and to a higher aggregation tendency for erythrocytes due to the degradation of the HES-coat and exposure of the polyplexes' positive charge. In vitro transfection experiments were conducted in 2 cell-lines±AA in the culture medium. The amylase-treated HES-decorated complexes showed up to 2 orders of magnitude higher transfection levels compared to the untreated HES-shielded particles, while AA had no effect on the transfection of PEG-coated or uncoated polyplexes. Finally, flow cytometry showed that the addition of AA increased the amount of delivered DNA per cell for the HES-shielded polyplexes. This study shows that decorating nanoparticles with HES can be a promising tool for the controlled shielding/deshielding of polyplexes. Copyright © 2012 Elsevier B.V. All rights reserved.

  19. HPLC profile, in vitro alpha-amylase, alpha-glucosidase inhibitory ...

    African Journals Online (AJOL)

    High Performance Liquid Chromatography profile revealed Resorcinol/Catechol to be one of the phenolics detected and may be responsible for GSEALE's inhibition of α-amylase and α-glucosidase and antioxidant properties. This shows that GS can probably be used for antidiabetic purpose due to significant antioxidant ...

  20. Glycinin, beta-conglycinin and alpha-amylase behaviour in artificially deteriorated and not deteriorated soybean

    OpenAIRE

    Salinas, Adriana Rita; Yoldjian, Ana Maria; Dietrich, María Lucrecia; Craviotto, Roque Mario; Bisaro, Vilma

    2002-01-01

    El objetivo del trabajo fue estudiar el comportamiento de glicinina y beta-conglicinina y la actividad de alfa-amilasa en semillas deterioradas y no deterioradas de 10 cultivares de soja [Glycine max (L.) Merr.]. Las semillas se sometieron a dos tratamientos: deterioradas por envejecimiento acelerado y no deterioradas. Se determinó la presencia de las proteínas de reserva a partir de semillas con 0, 3 y 8 días de germinadas por electroforesis en geles de poliacrilamida (SDS-PAGE). La activida...

  1. Salivary alpha-amylase and cortisol responsiveness following electrically stimulated physical stress in bipolar disorder patients

    Directory of Open Access Journals (Sweden)

    Tanaka Y

    2013-12-01

    Full Text Available Yoshihiro Tanaka, Yoshihiro Maruyama, Yoshinobu Ishitobi, Aimi Kawano, Tomoko Ando, Rie Ikeda, Ayako Inoue, Junko Imanaga, Shizuko Okamoto, Masayuki Kanehisa, Taiga Ninomiya, Jusen Tsuru, Jotaro Akiyoshi Department of Neuropsychiatry, Faculty of Medicine, Oita University, Hasama-Machi, Oita, Japan Background: Bipolar disorder (BP is often associated with a change in hypothalamus–pituitary–adrenal axis function change due to chronic stress. Salivary α-amylase (sAA levels increase in response to psychosocial stress and thus function as a marker of sympathoadrenal medullary system activity. However, sAA has been studied less often than salivary cortisol in BP patients. Method: We measured Profile of Mood States and State-Trait Anxiety Inventory scores, heart rate variability, and salivary cortisol levels during electrical stimulation stress in 25 BP patients and 22 healthy volunteers. Results: Tension–anxiety, depression–dejection, anger–hostility, fatigue, and confusion scores in BP patients significantly increased compared with those of the healthy controls. In contrast, the vigor scores of BP patients significantly decreased compared with those of the healthy controls. Significant difference in the sAA levels was observed between BP patients and healthy controls. sAA of female patients was significantly higher than that of female healthy controls, and sAA in male patients tended to be higher than that of male healthy controls. No difference in salivary cortisol was observed between BP patients and the healthy controls. Only three time points were measured before and after the electrical stimulation stress. Furthermore, sAA secretion by BP patients increased before and after electrical stimulation. Conclusion: These preliminary results suggest that sAA may be a useful biological marker for BP patients. Keywords: HPA axis, bipolar disorder, α-amylase, cortisol, SAM activity

  2. Occupational respiratory allergy with bakery workers : relationships with wheat and fungal [alpha]-amylase aeroallergen exposure

    NARCIS (Netherlands)

    Houba, R.

    1996-01-01

    In this thesis, results are presented of a cross-sectional epidemiological study among a few hundred bakery workers. Main focus was on the relationship between allergen exposure and the development of specific sensitization and respiratory allergy. Immunoassays were developed for measuring

  3. Rapid detection of fungal alpha-amylase in the work environment with a lateral flow immunoassay

    NARCIS (Netherlands)

    Bogdanovic, J.; Koets, M.; Sander, I.; Wouters, I.; Meijster, T.; Heederik, D.J.J.; Amerongen, van A.; Doekes, G.

    2006-01-01

    Background Occupational allergen exposure assessment usually requires airborne dust sampling at the worksite followed by dust extraction and enzyme immunoassay (EIA) analysis at the laboratory. Use of semiquantitative lateral flow immunoassays (LFIAs) may allow a more rapid detection procedure with

  4. Measuring Stress and Ability to Recover from Stress with Salivary Alpha-Amylase Levels

    Science.gov (United States)

    2011-04-01

    device and/or sleeve. Upon closing the lever , saliva is transferred from the test strip to the reagent paper. Concurrently, a timer begins when the... lever is closed and rings after the saliva transfer is complete. This process takes approximately 10 seconds. After the alarm rings, it is...resilience measurement system. Davis T., Lockhart T.E. (2003) The effects of age on stress and the biomechanics of slips and falls. In: Proceedings

  5. Brain alpha-amylase - a novel energy regulator important in Alzheimer disease?

    Science.gov (United States)

    Byman, Elin; Schultz, Nina; Fex, Malin; Wennström, Malin

    2018-02-27

    Reduced glucose metabolism and formation of polyglucosan bodies (PGB) are, beside amyloid beta plaques and neurofibrillary tangles, well-known pathological findings associated with Alzheimer's disease (AD). Since both glucose availability and PGB are regulated by enzymatic degradation of glycogen, we hypothesize that dysfunctional glycogen degradation is a critical event in AD progression. We therefore investigated whether alpha (α)-amylase, an enzyme known to efficiently degrade polysaccharides in the gastrointestinal tract, is expressed in the hippocampal CA1/subiculum and if the expression is altered in AD patients. Using immunohistochemical staining techniques, we show the presence of the α-amylase isotypes AMY1A and AMY2A in neuronal dendritic spines, pericytes and astrocytes. Moreover, AD patients showed reduced gene expression of α-amylase, but conversely increased protein levels of α-amylase as well as increased activity of the enzyme compared to non-demented controls. Lastly, we observed increased, albeit not significant, load of periodic acid-Schiff positive PGB in the brain of AD patients, which correlated with increased α-amylase activity. These findings show that α-amylase is expressed and active in the human brain, and suggest the enzyme to be affected, alternatively play a role, in the neurodegenerative Alzheimer's disease pathology. This article is protected by copyright. All rights reserved. © 2018 International Society of Neuropathology.

  6. Allele-specific primer based identification of dimeric alpha-amylase inhibitor

    OpenAIRE

    Pradeep Sharma; Pooja Sharma; Manoj Saini; S. S. Singh

    2011-01-01

    Wheat is one of the most important staple food crops cultivated over 200 mha in the range of environment throughout the world. Wheat production must continue to increase by 2% annually, more particularly in developing world including south-east Asia. Besides increasing the inherent productivity of wheat, it is important to minimize the losses caused to production by various abiotic and biotic factors. Alpha–amylase inhibitors are attractive candidates for the control of seed weevils as...

  7. Accuracy of alpha amylase in diagnosing microaspiration in intubated critically-ill patients.

    Directory of Open Access Journals (Sweden)

    Florent Dewavrin

    Full Text Available Amylase concentration in respiratory secretions was reported to be a potentially useful marker for aspiration and pneumonia. The aim of this study was to determine accuracy of α-amylase in diagnosing microaspiration in critically ill patients.Retrospective analysis of prospectively collected data collected in a medical ICU. All patients requiring mechanical ventilation for at least 48 h, and included in a previous randomized controlled trial were eligible for this study, provided that at least one tracheal aspirate was available for α-amylase measurement. As part of the initial trial, pepsin was quantitatively measured in all tracheal aspirates during a 48-h period. All tracheal aspirates were frozen, allowing subsequent measurement of α-amylase for the purpose of the current study. Microaspiration was defined as the presence of at least one positive tracheal aspirate for pepsin (>200 ng.mL-1. Abundant microaspiration was defined as the presence of pepsin at significant level in >74% of tracheal aspirates.Amylase was measured in 1055 tracheal aspirates, collected from 109 patients. Using mean α-amylase level per patient, accuracy of α-amylase in diagnosing microaspiration was moderate (area under the receiver operator curve 0.72±0.05 [95%CI 0.61-0.83], for an α-amylase value of 1685 UI.L-1. However, when α-amylase levels, coming from all samples, were taken into account, area under the receiver operator curve was 0.56±0.05 [0.53-0.60]. Mean α-amylase level, and percentage of tracheal aspirates positive for α-amylase were significantly higher in patients with microaspiration, and in patients with abundant microaspiration compared with those with no microaspiration; and similar in patients with microaspiration compared with those with abundant microaspiration. α-amylase and pepsin were significantly correlated (r2 = 0.305, p = 0.001.Accuracy of mean α-amylase in diagnosing microaspiration is moderate. Further, when all α-amylase levels were taken into account, α-amylase was inaccurate in diagnosing microaspiration, compared with pepsin.

  8. ON THE ACTIVITY OF alpha-AMYLASE IN SOME SPECIES OF ASIAN CYPRINIDS

    Directory of Open Access Journals (Sweden)

    Gabriela Vasile

    2006-08-01

    amylase in two Asian cyprinids namely, bighead carp (Aristichthys nobilis and silver carp (Hypophthalmichthys molitrix. The activity of -amylase has been determined by the Métais-Bieth method, the results obtained being expressed as mg starch / ml x 30 min. Our experimental data evidence some differences between the activity of -amylase from the digestive tube, in the two species under study.

  9. Effects of ultrasonication variables on the activity and properties of alpha amylase preparation.

    Science.gov (United States)

    Tran, Thi Thu Tra; Nguyen, Khanh Tien; Le, Van Viet Man

    2018-01-18

    Recently, ultrasound was demonstrated to increase enzyme activity in food industry. In most studies, an enzyme-substrate mixture was ultrasonicated. Very little is reported on the ultrasonication of enzyme preparation; the effects of ultrasonication variables on the enzyme activity and properties remained unclear. In this study, an α-amylase preparation was ultrasonicated under different conditions. At the ultrasonic frequency, power, temperature, and time of 20 kHz, 25 W/mL, 30°C, and 75 s, respectively, the secondary structure of the α-amylase was changed and that improved the catalytic activity by 47% over the control. The optimal temperature and pH of the α-amylase preparation did not change under the ultrasonic treatment. In addition, ultrasonic treatment increased kinetic parameters of the α-amylase including maximal velocity V max , Michaelis constant K m , turnover number K cat , and catalytic specificity constant K cat /K m . Nevertheless, the ultrasonicated α-amylase had lower thermal inactivation rate constant and half-life value than the nonultrasonicated enzyme. Ultrasonic treatment can be considered as a novel solution for improvement in enzyme activity. © 2018 American Institute of Chemical Engineers Biotechnol. Prog., 2018. © 2018 American Institute of Chemical Engineers.

  10. In-vitro cancer cell cytotoxicity and alpha amylase inhibition effect of seven tropical fruit residues

    Directory of Open Access Journals (Sweden)

    Priti Gupta

    2014-07-01

    Conclusions: Study suggests that these fruit residues demonstrate promising antidiabetic and anticancer activity that substantiated its ethno medicinal use and may provide new molecules for the treatment of these diseases.

  11. Alpha-amylase inhibitory activity and sterol composition of the marine algae, Sargassum glaucescens

    OpenAIRE

    Nasrin Payghami; Shahla Jamili; Abdolhossein Rustaiyan; Soodabeh Saeidnia; Marjan Nikan; Ahmad Reza Gohari

    2015-01-01

    Background: Sargassum species (phaeophyceae) are economically important brown algae in southern parts of Iran. Sargassum is mainly harvested as a row material in alginate production industries and is a source of plant foods or plant bio-stimulants even a component of animal foods. Objective: In this study, Sargassum glaucescens, collected from the seashore of Chabahar, was employed for phytochemical and biological evaluations. Materials and Methods: For that purpose, the dried algae was extra...

  12. Properties and applications of starch-converting enzymes of the alpha-amylase family

    NARCIS (Netherlands)

    van der Maarel, MJEC; van der Veen, B; Uitdehaag, JCM; Leemhuis, H; Dijkhuizen, L

    2002-01-01

    Starch is a major storage product of many economically important crops such as wheat, rice, maize, tapioca, and potato. A large-scale starch processing industry has emerged in the last century. In the past decades, we have seen a shift from the acid hydrolysis of starch to the use of

  13. 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.

  14. Noncontiguous finished genome sequences and description of Bacillus massiliglaciei, Bacillus mediterraneensis, Bacillus massilinigeriensis, Bacillus phocaeensis and Bacillus tuaregi, five new species identified by culturomics

    Directory of Open Access Journals (Sweden)

    F. Cadoret

    2017-09-01

    Full Text Available Microbial culturomics, which investigates microbial diversity by combining diversified culture conditions, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and 16S rDNA identification, allowed to identify five new species within the Bacillus genus. Bacillus massiliglaciei strain Marseille-P2600T, Bacillus mediterraneensis strain Marseille-P2384T, Bacillus massilinigeriensis strain Marseille-P2366T, Bacillus tuaregi strain Marseille-P2489T and Bacillus phocaeensis strain SIT16T are each the type strain of the corresponding bacterial species. These strains, the genomes of which are described here, are facultative anaerobic Gram-positive bacilli. Here, we describe the main characteristics of each bacterium and present their complete genome sequence and annotation.

  15. Bacillus pumilus SAFR-032 isolate

    Science.gov (United States)

    Venkateswaran, Kasthuri J. (Inventor)

    2007-01-01

    The present invention relates to discovery and isolation of a biologically pure culture of a Bacillus pumilus SAFR-032 isolate with UV sterilization resistant properties. This novel strain has been characterized on the basis of phenotypic traits, 16S rDNA sequence analysis and DNA-DNA hybridization. According to the results of these analyses, this strain belongs to the genus Bacillus. The GenBank accession number for the 16S rDNA sequence of the Bacillus pumilus SAFR-032 isolate is AY167879.

  16. Essential Bacillus subtilis genes

    DEFF Research Database (Denmark)

    Kobayashi, K.; Ehrlich, S.D.; Albertini, A.

    2003-01-01

    To estimate the minimal gene set required to sustain bacterial life in nutritious conditions, we carried out a systematic inactivation of Bacillus subtilis genes. Among approximate to4,100 genes of the organism, only 192 were shown to be indispensable by this or previous work. Another 79 genes were...... predicted to be essential. The vast majority of essential genes were categorized in relatively few domains of cell metabolism, with about half involved in information processing, one-fifth involved in the synthesis of cell envelope and the determination of cell shape and division, and one-tenth related...... to cell energetics. Only 4% of essential genes encode unknown functions. Most essential genes are present throughout a wide range of Bacteria, and almost 70% can also be found in Archaea and Eucarya. However, essential genes related to cell envelope, shape, division, and respiration tend to be lost from...

  17. Bacillus thuringiensis (Bt)

    Science.gov (United States)

    2004-01-01

    Bacillus thuringiensis (Bt), a natural bacteria found all over the Earth, has a fairly novel way of getting rid of unwanted insects. Bt forms a protein substance (shown on the right) that is not harmful to humans, birds, fish or other vertebrates. When eaten by insect larvae the protein causes a fatal loss of appetite. For over 25 years agricultural chemical companies have relied heavily upon safe Bt pesticides. New space based research promises to give the insecticide a new dimension in effectiveness and applicability. Researchers from the Consortium for Materials Development in Space along with industrial affiliates such as Abott Labs and Pern State University flew Bt on a Space Shuttle mission in the fall of 1996. Researchers expect that the Shuttle's microgravity environment will reveal new information about the protein that will make it more effective against a wider variety of pests.

  18. Measurement and Prediction of Protein Phase Behaviour and Protein-Protein Interactions

    DEFF Research Database (Denmark)

    Faber, Cornelius

    2006-01-01

    fra Bacillus halmapalus (BHA) og Bacillus licheniformis (BLA), som var blevet oprenset til teknisk grad. Udviklingen af optimerede mikrobielle stammer sammen med fremskreden gæringsteknologi fører til et renere industrielt gæringsmedie med højere produkt koncentration, som vil muliggøre en økonomisk...

  19. Bacillus velezensis is a later heterotypic synonym of Bacillus amyloliquefaciens.

    Science.gov (United States)

    Wang, Li-Ting; Lee, Fwu-Ling; Tai, Chun-Ju; Kuo, Hsiao-Ping

    2008-03-01

    Strain BCRC 14193, isolated from soil, shared more than 99 % 16S rRNA gene sequence similarity with Bacillus amyloliquefaciens BCRC 11601(T) and Bacillus velezensis BCRC 17467(T). This strain was previously identified as B. amyloliquefaciens, based on DNA-DNA hybridization, but its DNA relatedness value with B. velezensis BCRC 17467(T) was 89 %. To investigate the relatedness of strain BCRC 14193, B. amyloliquefaciens and B. velezensis, the partial sequence of the gene encoding the subunit B protein of DNA gyrase (gyrB) was determined. B. velezensis BCRC 17467(T) shared high gyrB gene sequence similarity with B. amyloliquefaciens BCRC 14193 (98.4 %) and all of the B. amyloliquefaciens strains available (95.5-95.6 %). DNA-DNA hybridization experiments revealed high relatedness values between B. velezensis BCRC 17467(T) and B. amyloliquefaciens BCRC 11601(T) (74 %) and the B. amyloliquefaciens reference strains (74-89 %). Based on these data and the lack of phenotypic distinctive characteristics, we propose Bacillus velezensis as a later heterotypic synonym of Bacillus amyloliquefaciens.

  20. Impacts of Bacillus thuringiensis var. israelensis and Bacillus ...

    African Journals Online (AJOL)

    The study assessed the impact of bio-larvicides- Bacillus thuringiensis var. israelensis (Bti) and B. sphaericus (Bs) on anopheline mosquito larval densities in four selected areas of Lusaka urban district. Larval densities were determined using a standard WHO protocol at each study area prior to and after larviciding.

  1. Phages Preying on Bacillus anthracis, Bacillus cereus, and Bacillus thuringiensis: Past, Present and Future

    Science.gov (United States)

    Gillis, Annika; Mahillon, Jacques

    2014-01-01

    Many bacteriophages (phages) have been widely studied due to their major role in virulence evolution of bacterial pathogens. However, less attention has been paid to phages preying on bacteria from the Bacillus cereus group and their contribution to the bacterial genetic pool has been disregarded. Therefore, this review brings together the main information for the B. cereus group phages, from their discovery to their modern biotechnological applications. A special focus is given to phages infecting Bacillus anthracis, B. cereus and Bacillus thuringiensis. These phages belong to the Myoviridae, Siphoviridae, Podoviridae and Tectiviridae families. For the sake of clarity, several phage categories have been made according to significant characteristics such as lifestyles and lysogenic states. The main categories comprise the transducing phages, phages with a chromosomal or plasmidial prophage state, γ-like phages and jumbo-phages. The current genomic characterization of some of these phages is also addressed throughout this work and some promising applications are discussed here. PMID:25010767

  2. Phages Preying on Bacillus anthracis, Bacillus cereus, and Bacillus thuringiensis: Past, Present and Future

    Directory of Open Access Journals (Sweden)

    Annika Gillis

    2014-07-01

    Full Text Available Many bacteriophages (phages have been widely studied due to their major role in virulence evolution of bacterial pathogens. However, less attention has been paid to phages preying on bacteria from the Bacillus cereus group and their contribution to the bacterial genetic pool has been disregarded. Therefore, this review brings together the main information for the B. cereus group phages, from their discovery to their modern biotechnological applications. A special focus is given to phages infecting Bacillus anthracis, B. cereus and Bacillus thuringiensis. These phages belong to the Myoviridae, Siphoviridae, Podoviridae and Tectiviridae families. For the sake of clarity, several phage categories have been made according to significant characteristics such as lifestyles and lysogenic states. The main categories comprise the transducing phages, phages with a chromosomal or plasmidial prophage state, γ-like phages and jumbo-phages. The current genomic characterization of some of these phages is also addressed throughout this work and some promising applications are discussed here.

  3. 76 FR 14289 - Bacillus thuringiensis

    Science.gov (United States)

    2011-03-16

    ... a plant- incorporated protectant in accordance with the terms of Experimental Use Permit (EUP) No... chemistry data for eCry3.1Ab were required for a human health effects assessment. Even so, preliminary... EPA granting registrations or experimental use permits of Bacillus thuringiensis-based pesticides or...

  4. Characterisation and profiling of Bacillus subtilis, Bacillus cereus and Bacillus licheniformis by MALDI-TOF mass fingerprinting.

    Science.gov (United States)

    Fernández-No, I C; Böhme, K; Díaz-Bao, M; Cepeda, A; Barros-Velázquez, J; Calo-Mata, P

    2013-04-01

    The Bacillus genus includes species such as Bacillus cereus, Bacillus licheniformis and Bacillus subtilis, some of which may be pathogenic or causative agents in the spoilage of food products. The main goal of this work was to apply matrix-assisted laser desorption ionisation-time of flight (MALDI-TOF) mass fingerprinting to the classification of these Bacillus species. Genetic analyses were also compared to phyloproteomic analyses. A collection of 57 Bacillus strains isolated from fresh and processed food and from culture collections were studied and their mass spectra compiled. The resulting mass fingerprints were compared and characteristic peaks at the strain and species levels were assigned. The results showed that MALDI-TOF was a good complementary approach to 16S rRNA sequencing and even a more powerful tool in the accurate classification of Bacillus species, especially for differentiating B. subtilis and B. cereus from Bacillus amyloliquefaciens and Bacillus thuringiensis, respectively. MALDI-TOF was also found to provide valuable information at both intra- and interspecies levels in the Bacillus species studied. Copyright © 2012 Elsevier Ltd. All rights reserved.

  5. Screening of Bacillus Species with Potentials of Antibiotics Production

    Directory of Open Access Journals (Sweden)

    Faruk Adamu KUTA

    2009-07-01

    Full Text Available Sixteen soil samples were collected from different refuse dump sites in Minna, the capital Niger State, and analysed for the presence of Bacillus species. Physical-chemical analysis of the soil samples revealed the followings: PH value 6.89-8.47; moisture content 1.58 – 21.21% and temperature 27-28ºC. Using both pour plate and streak method of inoculation, total bacterial count in the soil samples ranged from 3.8×104 cfu/g 16.0×104 cfu/g. The identified Bacillus species included: Bacillus cereus (30.8%, Bacillus brevis (1.9% Bacillus polymyxa (3.8%, Bacillus lichenifomis (13.5%, Bacillus spherericus (7.7%, Bacillus mycoides (13.5%, Bacillus pumilus (7.7%, Bacillus subtilis (3.8%, Bacillus alvei (1.9%, Bacillus laterosporous (1.9%, Bacillus firmus (9.6% and Bacillus circulars (3.8%. Antibiotic production tests indicated that nine Bacillus species out of twelve isolated in this study could be used to produce antibiotics that had effect on the test organisms. However, Bacillus polymyxa, Bacillus sphaericus and Bacillus laterosporous had little or no effect on the tested organisms. This study suggests that some Bacillus species have potential to produce high quality antibiotics that can be use to control microbial growth in future.

  6. FORMALDEHYDE GAS INACTIVATION OF BACILLUS ANTHRACIS, BACILLUS SUBTILIS AND GEOBACILLUS STEAROTHERMOPHILUS SPORES ON INDOOR SURFACE MATERIALS.

    Science.gov (United States)

    Research evaluated the decontamination of Bacillus anthracis, Bacillus subtilis, and Geobacillus stearothermophilus spores on indoor surface material using formaldehyde gas. Spores were dried on seven types of indoor surfaces and exposed to 1100 ppm formaldehyde gas for 10 hr. Fo...

  7. Culture conditions for the production of thermostable amylase by Bacillus sp Condições de cultura para produção de amilase termoestável por Bacillus sp

    Directory of Open Access Journals (Sweden)

    Carlos Eduardo de Souza Teodoro

    2000-10-01

    Full Text Available Studies on the alpha-amylase production were carried out with a bacterial strain isolated from a soil sample. The cells were cultivated in a mineral medium containing soluble starch as sole carbon source. The addition of calcium (10 mM or peptone (1% and yeast extract (0.5% to the mineral medium shortened the lag period and improved the growth and alpha-amylase synthesis. The addition of glucose to the culture diminished greatly the synthesis of alpha-amylase, demonstrating that a classical glucose effect is operative in this organism. The optimum temperature and initial medium pH for amylase synthesis by the organism were 50°C and 7.0 respectively. The optimal pH and temperature for activity were 6.0 and 50°C respectively. The enzyme extract retained 100% activity when incubated for one hour at 90°C and 40% at 60°C for 24 h. The addition of glucose to the culture diminished greatly the synthesis of alpha-amylase.Estudos sobre a produção de alfa-amilase foram realizados com uma bactéria isolada a partir de amostras de solo. As células foram cultivadas em um meio mineral contendo amido como única fonte de carbono. A adição de cálcio (10mM ou peptona (1,0% e extrato de levedura (0,5% ao meio mineral, encurtou o período lag, melhorou o crescimento e a síntese de alfa-amilase. A adição de glicose a cultura diminuiu grandemente a síntese de alfa-amilase, demonstrando que o efeito glicose ocorre neste microrganismo. A temperatura ótima e o pH inicial ótimo para a síntese da amilase pelo organismo foram 50ºC e 7,0 respectivamente. O pH e temperatura ótima para atividade foram 6,0 e 50ºC respectivamente. O extrato enzimático reteve 100% de atividade quando incubado por uma hora a 90ºC e 40% a 60ºC por 24 horas. A adição de glicose ao meio de cultura diminuiu grandemente a síntese de alfa-amilase.

  8. Characterization of microsatellite loci in the stick insects Bacillus rossius rossius, Bacillus rossius redtenbacheri and Bacillus whitei (Insecta : Phasmatodea)

    DEFF Research Database (Denmark)

    Andersen, DH; Pertoldi, C; Loeschcke, V

    2005-01-01

    Five microsatellite markers were obtained from a dinucleotide enriched genomic library of the stick insect Bacillus rossius rossius. The markers were tested in three species of Bacillus. All loci were polymorphic when tested across species. The number of alleles at each locus was low (maximum four...

  9. Pathogenomic sequence analysis of Bacillus cereus and Bacillus thuringiensis isolates closely related to Bacillus anthracis.

    Science.gov (United States)

    Han, Cliff S; Xie, Gary; Challacombe, Jean F; Altherr, Michael R; Bhotika, Smriti S; Brown, Nancy; Bruce, David; Campbell, Connie S; Campbell, Mary L; Chen, Jin; Chertkov, Olga; Cleland, Cathy; Dimitrijevic, Mira; Doggett, Norman A; Fawcett, John J; Glavina, Tijana; Goodwin, Lynne A; Green, Lance D; Hill, Karen K; Hitchcock, Penny; Jackson, Paul J; Keim, Paul; Kewalramani, Avinash Ramesh; Longmire, Jon; Lucas, Susan; Malfatti, Stephanie; McMurry, Kim; Meincke, Linda J; Misra, Monica; Moseman, Bernice L; Mundt, Mark; Munk, A Christine; Okinaka, Richard T; Parson-Quintana, B; Reilly, Lee Philip; Richardson, Paul; Robinson, Donna L; Rubin, Eddy; Saunders, Elizabeth; Tapia, Roxanne; Tesmer, Judith G; Thayer, Nina; Thompson, Linda S; Tice, Hope; Ticknor, Lawrence O; Wills, Patti L; Brettin, Thomas S; Gilna, Paul

    2006-05-01

    Bacillus anthracis, Bacillus cereus, and Bacillus thuringiensis are closely related gram-positive, spore-forming bacteria of the B. cereus sensu lato group. While independently derived strains of B. anthracis reveal conspicuous sequence homogeneity, environmental isolates of B. cereus and B. thuringiensis exhibit extensive genetic diversity. Here we report the sequencing and comparative analysis of the genomes of two members of the B. cereus group, B. thuringiensis 97-27 subsp. konkukian serotype H34, isolated from a necrotic human wound, and B. cereus E33L, which was isolated from a swab of a zebra carcass in Namibia. These two strains, when analyzed by amplified fragment length polymorphism within a collection of over 300 of B. cereus, B. thuringiensis, and B. anthracis isolates, appear closely related to B. anthracis. The B. cereus E33L isolate appears to be the nearest relative to B. anthracis identified thus far. Whole-genome sequencing of B. thuringiensis 97-27and B. cereus E33L was undertaken to identify shared and unique genes among these isolates in comparison to the genomes of pathogenic strains B. anthracis Ames and B. cereus G9241 and nonpathogenic strains B. cereus ATCC 10987 and B. cereus ATCC 14579. Comparison of these genomes revealed differences in terms of virulence, metabolic competence, structural components, and regulatory mechanisms.

  10. Isolation and characterization of cellulolytic Bacillus licheniformis ...

    African Journals Online (AJOL)

    Eight cellulose degrading bacteria were isolated from compost and were identified as Bacillus licheniformis by 16S rRNA sequencing. Among the eight isolates, Bacillus licheniformis B4, B7 and B8 showed the highest cellulase activity. B. licheniformis B4 and B8 showed the maximum cellulase activity during the stationary ...

  11. Isolat Bacillus Pelarut Fosfat dari Kalimas Surabaya

    OpenAIRE

    Zulaika, Enny; Ulfiyati, Nadia

    2015-01-01

    Ketersediaan fosfat terlarut di dalam tanah sangat terbatas karena kecenderungannya terikat dengan mineral tanah membentuk fosfat kompleks. Bakteri pelarut fosfat, salah satunya adalah Bacillus, dapat digunakan untuk membantu ketersediaan fosfat terlarut di dalam tanah sehingga dapat menggantikan pupuk fosfat. Tujuan penelitian adalah untuk mengetahui potensi isolat Bacillus spp. koleksi laboratorium Mikrobiologi dan Bioteknologi Biologi ITS Surabaya dalam melarutkan fosfat. Potensi pelarutan...

  12. Disinfection of Vegetative Cells of Bacillus anthracis

    Science.gov (United States)

    2016-03-01

    Society for Microbiology ; New Orleans, LA, 2004. American Public Health Association. Standard Methods for the Examination of Water and Wastewater...Disinfection kinetics of vegetative cells of Bacillus anthracis in water with free available chlorine ([FAC] 2 mg/L) and monochloramine ([MC] 2 mg/L) were...anthracis. Bacillus anthracis cells Drinking water Chlorine demand-free (CDF

  13. Real-Time PCR Assay for a Unique Chromosomal Sequence of Bacillus anthracis

    Science.gov (United States)

    2004-12-01

    13061 Neisseria lactamica .............................................................. 23970 Bacillus coagulans ...NEG Bacillus coagulane 7050 NEG NEG Bacillus cereus 13472 NEG NEG Bacillus licheniforms 12759 NEG NEG Bacillus cereus 13824 NEG NEG Bacillus ...Assay for a Unique Chromosomal Sequence of Bacillus anthracis Elizabeth Bode,1 William Hurtle,2† and David Norwood1* United States Army Medical

  14. Production dynamics of extracellular protease from Bacillus species ...

    African Journals Online (AJOL)

    ... showed that Bacillus species under study are good producers of extracellular protease at high temperature. This might be an indication that proteases produced would be thermostable. Keywords Protease; proteolytic bacteria; Bacillus macerans; Bacillus licheniformis; Bacillus subtilis. African Journal of Biotechnology Vol.

  15. Germination of Bacillus cereus spores : the role of germination receptors

    NARCIS (Netherlands)

    Hornstra, L.M.

    2007-01-01

    The Bacillus cereus sensu lato group forms a highly homogeneous subdivision of the genus Bacillus and comprises several species that are relevant for humans. Notorious is Bacillus anthracis, the cause of the often-lethal disease anthrax, while the insect pathogen Bacillus

  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. Free cortisol and salivary alpha-amylase levels during a six-hour-water immersion in healthy young men

    Science.gov (United States)

    Rohleder, N.; Wirth, D.; Fraßl, W.; Kowoll, R.; Schlemmer, M.; Vogler, S.; Kirsch, K. A.; Kirschbaum, C.; Gunga, H.-C.

    2005-08-01

    Limited data are available on the response of stress systems to microgravity. Increased activity of stress systems is reported during space flight, but unchanged or decreased activity during simulated microgravity. We here investigated the impact of head-out water immersion on the activity of the hypothalamus-pituitary-adrenal (HPA) axis and the sympathetic-adrenal-medullary (SAM) system.Eight healthy young men were exposed to a six-hour water immersion in a thermo neutral bath and a control condition. Saliva samples were taken before, during, and after interventions to assess cortisol as an index for HPA axis activity, and salivary α-amylase as an index for SAM system activity.Cortisol levels uniformly decreased during both conditions. Amylase levels increased during both conditions, but were significantly lower during the first half of water immersion compared to the control condition.In conclusion, the HPA axis is not influenced by simulated microgravity, while SAM system activity shows initial decreases during water immersion.

  18. 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.

  19. Evaluation of salivary catalase, vitamin C, and alpha-amylase in smokers and non-smokers: a retrospective cohort study.

    Science.gov (United States)

    Ahmadi-Motamayel, Fatemeh; Falsafi, Parisa; Goodarzi, Mohammad Taghi; Poorolajal, Jalal

    2017-05-01

    Saliva and its defence systems such as antioxidants and minerals are very important in the pathogenesis of different diseases. Cigarette smoking has many destructive effects. Oxidative stresses play an important role in the side effects of smoking. This study assessed the effect of cigarette smoking on salivary levels of catalase, vitamin C, and α-amylase. This retrospective cohort study was carried out in Hamadan, Iran, on 510 subjects; 259 subjects were smokers (the exposed group) and 251 were non-smokers (the unexposed group). Five microliters of unstimulated saliva was collected by spitting method. Catalase, vitamin C, and α-amylase salivary levels were determined by spectrophotometric assay. Data were analyzed with t-test using STATA 12. Vitamin C level in smokers was significantly lower than that in non-smokers. The salivary catalase levels were lower and α-amylase levels were higher in smokers, but the differences were not statistically significant (P = 0.416 and P = 0.265, respectively). Smokers were younger than non-smokers. Smoking resulted in a change in salivary antioxidant levels. Changes in antioxidant levels can influence the deleterious effects of smoking on oral mucosa; it might also indicate systemic changes and changes in the serum levels of oxidative agents. Further studies are necessary to understand the mechanisms and real effects of smoking, to determine the benefits of supplementary antioxidants for treatment and to reduce the dangerous side effects of smoking. © 2016 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  20. A chemically modified [alpha]-amylase with a molten-globule state has entropically driven enhanced thermal stability

    Energy Technology Data Exchange (ETDEWEB)

    Siddiqui, Khawar Sohail; Poljak, Anne; De Francisci, Davide; Guerriero, Gea; Pilak, Oliver; Burg, Dominic; Raftery, Mark J.; Parkin, Don M.; Trewhella, Jill; Cavicchioli, Ricardo (Sydney); (New South)

    2010-11-15

    The thermostability properties of TAA were investigated by chemically modifying carboxyl groups on the surface of the enzyme with AMEs. The TAA{sub MOD} exhibited a 200% improvement in starch-hydrolyzing productivity at 60 C. By studying the kinetic, thermodynamic and biophysical properties, we found that TAA{sub MOD} had formed a thermostable, MG state, in which the unfolding of the tertiary structure preceded that of the secondary structure by at least 20 C. The X-ray crystal structure of TAA{sub MOD} revealed no new permanent interactions (electrostatic or other) resulting from the modification. By deriving thermodynamic activation parameters of TAA{sub MOD}, we rationalised that thermostabilisation have been caused by a decrease in the entropy of the transition state, rather than being enthalpically driven. Far-UV CD shows that the origin of decreased entropy may have arisen from a higher helical content of TAA{sub MOD}. This study provides new insight into the intriguing properties of an MG state resulting from the chemical modification of TAA.

  1. 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

  2. The role of the enzyme alpha-amylase in binding of An(III)/Ln(III) by oral ingestion

    Energy Technology Data Exchange (ETDEWEB)

    Barkleit, A.; Bernhard, G. [Institute of Resource Ecology, Helmholtz-Zentrum Dresden-Rossendorf, P.O. Box 510119, 01314 Dresden (Germany); Division of Radiochemistry and Resource Ecology, Technische Universitaet Dresden, 01062 Dresden (Germany); Heller, A. [Institute of Resource Ecology, Helmholtz-Zentrum Dresden-Rossendorf, P.O. Box 510119, 01314 Dresden (Germany)

    2014-07-01

    In case of incorporation, radionuclides represent a serious health risk to humans due to their (radio-)toxicity. Thus, the determination of their speciation and transport on a molecular level is crucial for the understanding of the transport, metabolism, deposition and elimination in the human organisms. In case of oral ingestion of contaminated food or radioactive substances the first contact medium in the mouth is the aqueous bio-fluid saliva which contains inorganic ions (mainly Na{sup +}, K{sup +}, Ca{sup 2+}, Cl{sup -}, CO{sub 3}{sup 2-}, PO{sub 4}{sup 3-}) and numerous biomolecules, mainly proteins. One of the major proteins in saliva is the digestive enzyme α-amylase which catalyzes the hydrolysis of the α-1,4 glycosidic linkages of polysaccharides like starch or glycogen. [1] In this study the speciation of curium(III) and europium(III) in saliva as the first contact medium at oral incorporation was investigated with time-resolved laser-induced fluorescence spectroscopy (TRLFS). For TRLFS measurements, fresh saliva samples from human sources have been spiked in vitro with Eu(III) or Cm(III). The identification of the dominant species was achieved by a comparison of the spectroscopic data with reference spectra obtained from synthetic saliva and the main single components of the bio-fluid. In the pH range from 6.8 to 7.4 similar spectra were obtained. With respect to reference data, the spectra indicate the formation of a ternary metal complex containing phosphate and carbonate anions and, in addition, a coordination of organic matter, namely α-amylase, to the central metal cation is suggested. To get more information about the binding behavior of α-amylase various investigations with Eu(III) as inactive analog for An(III) were carried out with porcine pancreatic α-amylase (PPA) which serves as model system for various α-amylase species. Sorption experiments showed a high affinity of Eu(III) to α-amylase in a wide pH range, namely between pH 4 and 8. The analysis of binding isotherms demonstrated that up to 3 Eu{sup 3+} ions are bound to one enzyme molecule. Hence, the Eu{sup 3+} ions seem to replace the Ca{sup 2+} ions, a well-known mechanism in biological systems. The effect of Eu{sup 3+} on enzyme activity was determined with the α-amylase assay method by Bernfeld [2]. Eu{sup 3+} shows a strong inhibition effect on the enzyme activity, but in the presence of Ca{sup 2+} in excess the enzyme activity remains nearly unaffected. This effect might be useful for the refinement of decontamination strategies. (authors)

  3. 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....

  4. In vitro antibacterial, alpha-amylase inhibition potential of three nudibranchs extracts from South East coast of India

    Directory of Open Access Journals (Sweden)

    Giji Sadhasivam

    2013-10-01

    Full Text Available Objective: To study the antibacterial and antiamylase properties of methanol and acetone extracts of nudibranchs including Bursatella leachii (B. leachii, Kalinga ornata (K. ornata, Aplysia sp. Methods: Crude methanol and acetone extracts of sea slugs were tested for inhibition of fish bacterial pathogens' growth through disc diffusion method. The activity was measured based on the formation of inhibition zone around the disc impregnated with crude extracts. The α-amylase inhibitory effect was also measured calorimetrically. The chemical fingerprinting of the extract was recorded with HPTLC and GC-MS. Results: The solvent extracts of all the three sea slugs showed antibacterial property. The maximum zone of inhibition (>15-20 mm was recorded for methanol and acetone extracts of K. ornata. The methanol extract of Aplysia sp. exhibited 93% inhibition against α-amylase, following by B. leachii (methanol 70.6% and K. ornata (methanol 49.03% inhibition respectively. The acetone extracts didn' t show any notable inhibition. The presence of free amino acids like lysine, aspartic acid, glutamic acid, arginine etc., terpenoids and pigents were confirmed through HPTLC analysis. The presence of siloxanes and propanoic acid were also revealed through GC-MS. Conclusions: This study suggests that further scrutinisation of the B. leachii, K. ornata and Aplysia sp. will pave the way for development of antibacterial and α-amylase inhibitory agent for therapeutic application.

  5. Catalytic mechanism and product specificity of cyclodextrin glycosyltransferase, a prototypical transglycosylase from the alpha-amylase family

    NARCIS (Netherlands)

    Uitdehaag, JCM; van der Veen, BA; Dijkhuizen, L; Dijkstra, BW

    2002-01-01

    The catalytic mechanism of cyclodextrin glycosyltransferase, a member of the a-amylase family, is reviewed. The focus is put on the bond cleavage mechanism, the nature of the transition state and of the covalent intermediate, and on the stereo-electronic and lateral protonation contributions to

  6. In vitro and in vivo inhibition of alpha-amylases of stored-product mite Acarus siro

    Czech Academy of Sciences Publication Activity Database

    Hubert, J.; Marešová, Lucie; Hýblová, Jana; Kudlíková, I.; Stejskal, V.; Mareš, Michael

    2005-01-01

    Roč. 35, - (2005), s. 281-291 ISSN 0168-8162 R&D Projects: GA MŠk(CZ) OC 842.20; GA ČR GP203/02/P081 Institutional research plan: CEZ:AV0Z4055905 Keywords : Acarus siro * inhibitor * stored-product mite Subject RIV: CE - Biochemistry Impact factor: 0.978, year: 2005

  7. Adolescents' Increasing Stress Response to Social Evaluation: Pubertal Effects on Cortisol and Alpha-Amylase during Public Speaking

    Science.gov (United States)

    van den Bos, Esther; de Rooij, Mark; Miers, Anne C.; Bokhorst, Caroline L.; Westenberg, P. Michiel

    2014-01-01

    Stress responses to social evaluation are thought to increase during adolescence, which may be due to pubertal maturation. However, empirical evidence is scarce. This study is the first to investigate the relation between pubertal development and biological responses to a social-evaluative stressor longitudinally. Participants performed the Leiden…

  8. Gedunin and Azadiradione: Human Pancreatic Alpha-Amylase Inhibiting Limonoids from Neem (Azadirachta indica as Anti-Diabetic Agents.

    Directory of Open Access Journals (Sweden)

    Sudha Ponnusamy

    Full Text Available Human pancreatic α-amylase (HPA inhibitors offer an effective strategy to lower postprandial hyperglycemia via control of starch breakdown. Limonoids from Azadirachta indica known for their therapeutic potential were screened for pancreatic α-amylase inhibition, a known anti-diabetic target. Studies were carried out to reveal their mode of action so as to justify their hypoglycemic potential. Of the nine limonoids isolated/semi-synthesized from A.indica and screened for α-amylase inhibition, azadiradione and exhibited potential inhibition with an IC50 value of 74.17 and 68.38 μM, respectively against HPA under in vitro conditions. Further screening on AR42J α-amylase secretory cell line for cytotoxicity and bioactivity revealed that azadiradione and gedunin exhibited cytotoxicity with IC50 of 11.1 and 13.4μM. Maximal secreted α-amylase inhibition of 41.8% and 53.4% was seen at 3.5 and 3.3μM, respectively. Michaelis-Menten kinetics suggested a mixed mode of inhibition with maltopentaose (Ki 42.2, 18.6 μM and starch (Ki' 75.8, 37.4 μM as substrate with a stiochiometry of 1:1 for both azadiradione and gedunin, respectively. The molecular docking simulation indicated plausible π-alkyl and alkyl-alkyl interactions between the aromatic amino acids and inhibitors. Fluorescence and CD confirmed the involvement of tryptophan and tyrosine in ligand binding to HPA. Thermodynamic parameters suggested that binding is enthalpically and entropically driven with ΔG° of -21.25 kJ mol-1 and -21.16 kJ mol-1 for azadiradione and gedunin, respectively. Thus, the limonoids azadiradione and gedunin could bind and inactivate HPA (anti-diabetic target and may prove to be lead drug candidates to reduce/control post-prandial hyperglycemia.

  9. Inhibitory effect of leaf extract of Newbouldia laevis on the metabolic activities of alpha-glucosidase and alpha-amylase

    Directory of Open Access Journals (Sweden)

    Oyetunji T. Kolawole

    2013-12-01

    Full Text Available In this study, the effect of ethanol extract of the leaves of Newbouldia laevis on the activity of α-amylase and α-glucosidase was investigated. Inhibitory effect of N. laevis extract on α-glucosidase was tested in vitro using baker’s yeast α-glucosidase and rat intestinal α-glucosidase while α-amylase inhibitory effect was assayed using rat pancreatic α-amylase. α-Glucosidase inhibitory effect of the extract was also tested in vivo in diabetic and non-diabetic rats. N. laevis extract exhibited good α-glucosidase inhibitory activity in vitro with IC50 values of 2.2 µg/mL and 43.5 µg/mL for baker’s yeast and rat intestinal α-glucosidase respectively. The extract also inhibited rat pancreatic α-amylase activity with IC50 value of 58.7 µg/mL. In both diabetic and non-diabetic rats, N. laevis extract caused a significant reduction in postprandial blood glucose level after oral sucrose load. The results of this study indicate that N. laevis extract exerts its glucose-lowering effect through inhibition of α-glucosidase and α-amylase.

  10. Phylogenetic Distribution of Intron Positions in Alpha-Amylase Genes of Bilateria Suggests Numerous Gains and Losses

    OpenAIRE

    Da Lage, Jean-Luc; Maczkowiak, Frédérique; Cariou, Marie-Louise

    2011-01-01

    Most eukaryotes have at least some genes interrupted by introns. While it is well accepted that introns were already present at moderate density in the last eukaryote common ancestor, the conspicuous diversity of intron density among genomes suggests a complex evolutionary history, with marked differences between phyla. The question of the rates of intron gains and loss in the course of evolution and factors influencing them remains controversial. We have investigated a single gene family, al...

  11. Effect of Ultrasonic Waves on the Heat Resistance of Bacillus cereus and Bacillus licheniformis Spores

    Science.gov (United States)

    Burgos, J.; Ordóñez, J. A.; Sala, F.

    1972-01-01

    Heat resistance of Bacillus cereus and Bacillus licheniformis spores in quarter-strength Ringer solution decreases markedly after ultrasonic treatments which are unable to kill a significant proportion of the spore population. This effect does not seem to be caused by a loss of Ca2+ or dipicolinic acid. The use of ultrasonics to eliminate vegetative cells or to break aggregates in Bacillus spore suspensions to be used subsequently in heat resistance experiments appears to be unadvisable. PMID:4627969

  12. Statistically designed optimisation of enzyme catalysed starch removal from potato pulp

    DEFF Research Database (Denmark)

    Thomassen, Lise Vestergaard; Meyer, Anne S.

    2010-01-01

    and temperature on the amount of starch released from the potato pulp. The data demonstrated that all the starch could be released from potato pulp in one step when 8% (w/w) dry potato pulp was treated with 0.2% (v/w) (enzyme/substrate (E/S)) of a thermostable Bacillus licheniformis alpha-amylase (Termamyl(R) SC...

  13. Bacillus amyloliquefaciens, Bacillus velezensis, and Bacillus siamensis Form an ?Operational Group B. amyloliquefaciens? within the B. subtilis Species Complex

    OpenAIRE

    Fan, Ben; Blom, Jochen; Klenk, Hans-Peter; Borriss, Rainer

    2017-01-01

    The plant growth promoting model bacterium FZB42T was proposed as the type strain of Bacillus amyloliquefaciens subsp. plantarum (Borriss et al., 2011), but has been recently recognized as being synonymous to Bacillus velezensis due to phylogenomic analysis (Dunlap C. et al., 2016). However, until now, majority of publications consider plant-associated close relatives of FZB42 still as “B. amyloliquefaciens.” Here, we reinvestigated the taxonomic status of FZB42 and related strains in its con...

  14. Bacillus and biopolymer: Prospects and challenges

    Directory of Open Access Journals (Sweden)

    Swati Mohapatra

    2017-12-01

    Full Text Available The microbially derived polyhydroxyalkanoates biopolymers could impact the global climate scenario by replacing the conventional non-degradable, petrochemical-based polymer. The biogenesis, characterization and properties of PHAs by Bacillus species using renewable substrates have been elaborated by many for their wide applications. On the other hand Bacillus species are advantageous over other bacteria due to their abundance even in extreme ecological conditions, higher growth rates even on cheap substrates, higher PHAs production ability, and the ease of extracting the PHAs. Bacillus species possess hydrolytic enzymes that can be exploited for economical PHAs production. This review summarizes the recent trends in both non-growth and growth associated PHAs production by Bacillus species which may provide direction leading to future research towards this growing quest for biodegradable plastics, one more critical step ahead towards sustainable development.

  15. Characterization of 21 Strains of Bacillus Anthracis

    National Research Council Canada - National Science Library

    Kournikakis, B

    2000-01-01

    Twenty-one strains of Bacillus anthracis currently held in the culture collection at DRES were characterized by colonial morphology, antibiotic sensitivity and BiologTM metabolic identification profiles...

  16. Antimicrobial effect of lactobacillus and bacillus derived ...

    African Journals Online (AJOL)

    This study focused on the screening, production, extraction of biosurfactants from Lactobacillus and Bacillus bacteria and their antimicrobial properties against causal microorganisms of food borne infections (food borne pathogens). The biosurfactants were investigated for potential antimicrobial activity using disk diffusion.

  17. Cell Physiology and Protein Secretion of Bacillus licheniformis Compared to Bacillus subtilis

    NARCIS (Netherlands)

    Voigt, Birgit; Antelmann, Haike; Albrecht, Dirk; Ehrenreich, Armin; Maurer, Karl-Heinz; Evers, Stefan; Gottschalk, Gerhard; van Dijl, Jan Maarten; Schweder, Thomas; Hecker, Michael

    2009-01-01

    The genome sequence of Bacillus subtilis was published in 1997 and since then many other bacterial genomes have been sequenced, among them Bacillus licheniformis in 2004. B. subtilis and B. licheniformis are closely related and feature similar saprophytic lifestyles in the soil. Both species can

  18. Bacillus velezensis is not a later heterotypic synonym of Bacillus amyloliquefaciens; Bacillus methylotrophicus, Bacillus amyloliquefaciens subsp. plantarum and 'Bacillus oryzicola' are later heterotypic synonyms of Bacillus velezensis based on phylogenomics.

    Science.gov (United States)

    Dunlap, Christopher A; Kim, Soo-Jin; Kwon, Soon-Wo; Rooney, Alejandro P

    2016-03-01

    Bacillus velezensis was previously reported to be a later heterotypic synonym of Bacillus amyloliquefaciens , based primarily on DNA-DNA relatedness values. We have sequenced a draft genome of B. velezensis NRRL B-41580 T . Comparative genomics and DNA-DNA relatedness calculations show that it is not a synonym of B. amyloliquefaciens. It was instead synonymous with Bacillus methylotrophicus. ' Bacillus oryzicola ' is a recently described species that was isolated as an endophyte of rice ( Oryza sativa ). The strain was demonstrated to have plant-pathogen antagonist activity in greenhouse assays, and the 16S rRNA gene was reported to have 99.7 % sequence similarity with Bacillus siamensis and B. methylotrophicus , which are both known for their plant pathogen antagonism. To better understand the phylogenetics of these closely related strains, we sequenced the genome of ' B . oryzicola ' KACC 18228. Comparative genomic analysis showed only minor differences between this strain and the genomes of B. velezensis NRRL B-41580 T , B. methylotrophicus KACC 13015 T and Bacillus amyloliquefaciens subsp. plantarum FZB42 T . The pairwise in silico DNA-DNA hybridization values calculated in comparisons between the strains were all greater than 84 %, which is well above the standard species threshold of 70 %. The results of morphological, physiological, chemotaxonomic and phylogenetic analyses indicate that the strains share phenotype and genotype coherence. Therefore, we propose that B. methylotrophicus KACC 13015 T , B. amyloliquefaciens subsp. plantarum FZB42 T , and ' B. oryzicola' KACC 18228 should be reclassified as later heterotypic synonyms of B. velezensis NRRL B-41580 T , since the valid publication date of B. velezensis precedes the other three strains.

  19. Production of amylolytic enzymes by bacillus spp

    International Nuclear Information System (INIS)

    Dawood, Elham Shareif

    1997-12-01

    Sixty six bacteria and twenty fungi were isolated from various sources. These varied from rotten fruites to local drinks and soil samples from different parts of Sudan. On the basis of index of amylolytic activity, forty one bacteria and twelve fungi were found to hydrolyse strach. The best ten strach hydrolysing isolates were identified all as bacilli (Bacillus licheniformis SUD-K 1 , SUD-K 2 , SUD-K 4 , SUD-O, SUD-SRW, SUD-BRW, SUD-By, Bacillus subtilis SUD-K 3 , and Bacillus circulans SUD-D and SUD-K 7 ). Their amylase productivity was studied with respect to temperature and time. Amylolytic activity was measured by spectrophotometer, the highest activity was produced in around 24 hours of growth in all; six of which gave the highest amylase activity at 50 deg C and the rest at 45C. Based on the thermal production six isolates were chosen for further investigation. These were Bacillus licheniformis SUD-K 1 , SUD-K 2 , SUD-K 4 , SUD-O, Bacillus subtilis SUD-K 3 and Bacillus circulans SUD-K 7 . The inclusion of strach and Mg ++ ions in the culture medium gave the highest enzyme yield. The Ph 9.0 was found to be the optimum for amylase production for all isolates except Bacillus subtilis SUD-K 3 which had an optimum at pH 7.0. Three isolates (Bacillus licheniformis SUD-K 1 , SUD-K 4 and SUD-O recorded highestamylase production in a medium supplemented with peptone while the rest (Bacillus licheniformis SUD-K 2 , Bacillus subtilis SUD-K 3 and Bacillus circulans SUD-K 7 ) gave highest amylase productivity in a medium supplemented with malt extract. Four isolates (Bacillus licheniformis SUD-K 1 and Bacillus subtilis SUD-K 3 gave maximum amylase production in a medium containing 0.5% soluble strach while the rest (gave maximum amylase production at 2%. Soluble strach was found to be best substrate among the different carbon sources tested. The maximum temperature for amylase activity ranged from 60-70 deg C and 1% strach concentration was optimum for all isolates

  20. Measurement of Metabolic Activity in Dormant Spores of Bacillus Species

    Science.gov (United States)

    2015-01-14

    SECURITY CLASSIFICATION OF: Spores of Bacillus megaterium and Bacillus subtilis were harvested shortly after release from sporangia, incubated under...Dec-2014 Approved for Public Release; Distribution Unlimited Final Report: Measurement of Metabolic Activity in Dormant Spores of Bacillus Species...Research Office P.O. Box 12211 Research Triangle Park, NC 27709-2211 spores, Bacillus , spore dormancy, 3-phosphoglycerate REPORT DOCUMENTATION PAGE 11

  1. DECONTAMINATION ASSESSMENT OF BACILLUS ANTHRACIS, BACILLUS SUBTILIS, AND GEOBACILLUS STEAROTHERMOPHILUS SPORES ON INDOOR SURFACTS USING A HYDROGEN PERIOXIDE GAS GENERATOR

    Science.gov (United States)

    Aims: To evaluate the decontamination of Bacillus anthracis, Bacillus subtilis, and Geobacillus stearothermophilus spores on indoor surface materials using hydrogen peroxide gas. Methods and Results: B. anthracis, B. subtilis, and G. Stearothermophilus spores were dried on seven...

  2. 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 ...

  3. BOOK REVIEW – BACILLUS THURINGIENSIS: A CORNERSTONE OF MODERN AGRICULTURE BACILLUS THURINGIENSIS

    Science.gov (United States)

    Are you interested in the technical issues surrounding the use of Bacillus thuringiensis pesticidal traits as sprays and as plant incorporated protectants (transgenic crops)? Should the dimensions of human health, ecology, entomology, risk assessment, resistance management, and d...

  4. Bacillus amyloliquefaciens, Bacillus velezensis, and Bacillus siamensis Form an "Operational Group B. amyloliquefaciens" within the B. subtilis Species Complex.

    Science.gov (United States)

    Fan, Ben; Blom, Jochen; Klenk, Hans-Peter; Borriss, Rainer

    2017-01-01

    The plant growth promoting model bacterium FZB42 T was proposed as the type strain of Bacillus amyloliquefaciens subsp. plantarum (Borriss et al., 2011), but has been recently recognized as being synonymous to Bacillus velezensis due to phylogenomic analysis (Dunlap C. et al., 2016). However, until now, majority of publications consider plant-associated close relatives of FZB42 still as " B. amyloliquefaciens ." Here, we reinvestigated the taxonomic status of FZB42 and related strains in its context to the free-living soil bacterium DSM7 T , the type strain of B. amyloliquefaciens . We identified 66 bacterial genomes from the NCBI data bank with high similarity to DSM7 T . Dendrograms based on complete rpoB nucleotide sequences and on core genome sequences, respectively, clustered into a clade consisting of three tightly linked branches: (1) B. amyloliquefaciens , (2) Bacillus siamensis , and (3) a conspecific group containing the type strains of B. velezensis, Bacillus methylotrophicus , and B. amyloliquefaciens subsp. plantarum . The three monophyletic clades shared a common mutation rate of 0.01 substitutions per nucleotide position, but were distantly related to Bacillus subtilis (0.1 substitutions per nucleotide position). The tight relatedness of the three clusters was corroborated by TETRA, dDDH, ANI, and AAI analysis of the core genomes, but dDDH and ANI values were found slightly below species level thresholds when B. amyloliquefaciens DSM7 T genome sequence was used as query sequence. Due to these results, we propose that the B. amyloliquefaciens clade should be considered as a taxonomic unit above of species level, designated here as "operational group B. amyloliquefaciens " consisting of the soil borne B. amyloliquefaciens , and plant associated B. siamensis and B. velezensis , whose members are closely related and allow identifying changes on the genomic level due to developing the plant-associated life-style.

  5. Comparative genome analysis of Bacillus cereus group genomes withBacillus subtilis

    Energy Technology Data Exchange (ETDEWEB)

    Anderson, Iain; Sorokin, Alexei; Kapatral, Vinayak; Reznik, Gary; Bhattacharya, Anamitra; Mikhailova, Natalia; Burd, Henry; Joukov, Victor; Kaznadzey, Denis; Walunas, Theresa; D' Souza, Mark; Larsen, Niels; Pusch,Gordon; Liolios, Konstantinos; Grechkin, Yuri; Lapidus, Alla; Goltsman,Eugene; Chu, Lien; Fonstein, Michael; Ehrlich, S. Dusko; Overbeek, Ross; Kyrpides, Nikos; Ivanova, Natalia

    2005-09-14

    Genome features of the Bacillus cereus group genomes (representative strains of Bacillus cereus, Bacillus anthracis and Bacillus thuringiensis sub spp israelensis) were analyzed and compared with the Bacillus subtilis genome. A core set of 1,381 protein families among the four Bacillus genomes, with an additional set of 933 families common to the B. cereus group, was identified. Differences in signal transduction pathways, membrane transporters, cell surface structures, cell wall, and S-layer proteins suggesting differences in their phenotype were identified. The B. cereus group has signal transduction systems including a tyrosine kinase related to two-component system histidine kinases from B. subtilis. A model for regulation of the stress responsive sigma factor sigmaB in the B. cereus group different from the well studied regulation in B. subtilis has been proposed. Despite a high degree of chromosomal synteny among these genomes, significant differences in cell wall and spore coat proteins that contribute to the survival and adaptation in specific hosts has been identified.

  6. tolerant alkaline protease from Bacillus coagulans PSB

    African Journals Online (AJOL)

    oyaide

    2013-05-22

    May 22, 2013 ... optimum activity at 60°C and pH 8.0 with casein as substrate. The enzyme was .... appropriate buffers. 50 mM of buffer solutions (sodium citrate, pH .... Table 2. Hydrolysis of protein substrates by protease from Bacillus coagulans PSB-07. Substrate. Relative activity (%). Casein. 100. Gelatin. 18. BSA. 72.

  7. Probiotic Bacillus species and Saccharomyces boulardii improve ...

    African Journals Online (AJOL)

    Probiotic Bacillus species and Saccharomyces boulardii improve performance, gut histology and immunity in broiler chickens. ... Carcass yield, liver weights, breast muscle values, and abdominal fat weights were reduced in groups fed with 100 or 150 g/ton of Microguard. Caecal coliforms, Salmonella and Escherichia coli ...

  8. Complete Genome of Bacillus megaterium Podophage Pascal.

    Science.gov (United States)

    Snowden, Jeffery D; Vega Gonzalez, Alexander E; Maroun, Justin W; Hernandez, Adriana C; Kuty Everett, Gabriel F

    2015-01-29

    Podophage Pascal infects Bacillus megaterium, a commonly used model organism in biochemical research and an important industrial-scale protein production system. Here, we report the sequenced and annotated genome of Pascal and describe its prominent features. Bacteriophages such as Pascal may be valuable tools for research and industry. Copyright © 2015 Snowden et al.

  9. Antibacterial potential components of Bacillus species and ...

    African Journals Online (AJOL)

    Honey is a sweet viscous liquid produced by honey bee, Apis mellifera from the nectar of plants. Honey is a natural product that has been used from ancient times till now as food and for medicinal purpose. This study was carried out to determine the mode of action of Bacillus species and antibiotics residues in branded and ...

  10. Bacillus subtilis Spore Inner Membrane Proteome

    NARCIS (Netherlands)

    Zheng, L.; Abhyankar, W.; Ouwerling, N.; Dekker, H.L.; van Veen, H.; van der Wel, N.N.; Roseboom, W.; de Koning, L.J.; Brul, S.; de Koster, C.G.

    2016-01-01

    The endospore is the dormant form of Bacillus subtilis and many other Firmicutes. By sporulation, these spore formers can survive very harsh physical and chemical conditions. Yet, they need to go through germination to return to their growing form. The spore inner membrane (IM) has been shown to

  11. Protein-Tyrosine Phosphorylation in Bacillus subtilis

    DEFF Research Database (Denmark)

    Mijakovic, Ivan; Petranovic, Dina; Bottini, N.

    2005-01-01

    phosphorylation, indicating that this post-translational modifi cation could regulate physiological processes ranging from stress response and exopolysaccharide synthesis to DNA metabolism. Some interesting work in this fi eld was done in Bacillus subtilis , and we here present the current state of knowledge...

  12. strains of pseudomonas aeruginosa and bacillus cereus

    African Journals Online (AJOL)

    DR. AMINU

    DETERMINATION OF THE GENETIC MARKER OF THE MUTAGENIZED. STRAINS OF PSEUDOMONAS AERUGINOSA AND BACILLUS CEREUS. ISOLATED FROM EFFLUENT OF PETROLEUM REFINERY. Idise, O. E.1, Ameh, J.B.2 Yakubu, S.E. 2, Okuofu, C.A. 3 and Ado, S.A.2. 1 Department of Microbiology, Delta ...

  13. Preliminary investigations reveal that Bacillus thuringiensis δ ...

    African Journals Online (AJOL)

    The imminent introduction of transgenic crops into Kenya requires a rigorous assessment of the potential risks involved. This study focused on the possible effect of Bacillus thuringiensisδ-endotoxin [CryIA(c)] on arbuscular mycorrhizal fungi (AMF) associated with sorghum. In green house experiments, sorghum seedlings ...

  14. Isolation and characterization of native Bacillus thuringiensis ...

    African Journals Online (AJOL)

    ... (<30% mortality) or non insecticidal activity. However, results of motility, hemolytic activity, antibiotic-susceptibility patterns, and crystal shape, seem to suggest that many of our Bt isolates may exhibit parasporins activity. Key words: Bacillus thuringiensis, parasporal crystal, isolation, biochemical type, insecticidal, cry gene, ...

  15. Purification And Characterization Of Marine Bacillus Thuringiensis ...

    African Journals Online (AJOL)

    Urease was purified to homogeneity from Bacillus thuringiensis N2 using different purification steps namely, 55% acetone precipitation, DEAE-Sephadex A50 anion exchange column and Sephadex G120-200 gel filtration chromatography. The enzyme was purified 95.27 fold and showed a final specific activity of 10.48 ...

  16. Molecular characterization of Lepidopteran specific Bacillus ...

    African Journals Online (AJOL)

    Bacillus thuringiensis (Bt) strains pathogenic to Lepidopteran insects and native to hilly zone soils of Karnataka (India) were explored. 19 strains were isolated from the soils and identified by morphological and microscopic characters. Toxicity level of the Bt isolates was tested by treating third Instar larvae of silkworm ...

  17. Molecular characterization of Lepidopteran specific Bacillus ...

    African Journals Online (AJOL)

    Guest

    2013-05-15

    May 15, 2013 ... Department of Plant Biotechnology, University of Agricultural Sciences, GKVK Campus, Bangalore, India. Accepted 24 April, 2013. Bacillus thuringiensis (Bt) strains pathogenic to Lepidopteran insects and native to hilly zone soils of. Karnataka ... In one of the isolates (Bt9), the cry gene was not detected.

  18. CASE REPORT Uncommon Pathogen Bacillus Cereus Causing ...

    African Journals Online (AJOL)

    2018-01-01

    Jan 1, 2018 ... Uncommon Pathogen Bacillus Cereus Causing Subdural Empyema in a Child. Prastiya Indra Gunawan1*, Leny Kartina1, Dwiyanti Puspitasari1, Erny Erny2. OPEN ACCESS ... secondary to middle ear infection, meningitis, brain surgery, ... classic clinical syndrome is an acute febrile illness punctuated by.

  19. Antimicrobials of Bacillus species: mining and engineering

    NARCIS (Netherlands)

    Zhao, Xin

    2016-01-01

    Bacillus sp. have been successfully used to suppress various bacterial and fungal pathogens. Due to the wide availability of whole genome sequence data and the development of genome mining tools, novel antimicrobials are being discovered and updated,;not only bacteriocins, but also NRPs and PKs. A

  20. Exopolysaccharide production by Bacillus subtilis NCIM 2063 ...

    African Journals Online (AJOL)

    Three bacterial strains, Bacillus subtilis NCIM 2063, Pseudomonas aeruginosa NCIM 2862 and Streptococcus mutans MTCC 1943 were examined for their exopolysaccharide (EPS) producing ability at the laboratory level. Basal salts solution (BSS), minimal salts medium (MSM), nitrogen free medium (NFM), chemically ...

  1. CASE REPORT Uncommon Pathogen Bacillus Cereus Causing ...

    African Journals Online (AJOL)

    2018-01-01

    Jan 1, 2018 ... A complete blood count showed white blood cell count of 13.800/mm3 and the CRP level was 8.3 mg/L. Craniotomy following burr hole drainage procedure was performed to decrease intracranial pressure. The liquor culture indicated Bacillus cereus. A meropenem injection and metronidazole infusion ...

  2. Utilization of corn starch as sustrate for ß-Amylase by Bacillus SPP

    African Journals Online (AJOL)

    Corn starch was used as substrate for ß -amylase production from ten(10) amylolytic species of the genus Bacillus isolated locally from soil, waste water and food sources. Ten bacillus strains was made up of two strains each of Bacillus macerans, Bacillus licheniformis and Bacillus circulans. Also included are B. coagulans, ...

  3. L-Glutamic acid production by Bacillus spp. isolated from vegetable ...

    African Journals Online (AJOL)

    Ogiri” (fermented vegetable proteins) in Nigeria. The isolates were identified as Bacillus subtilis (6), (27.3%), Bacillus pumilus (5), (22.7%), Bacillus licheniformis (5), (27.3%) and Bacillus polymyxa (6), (22.7%). Four species of the Bacillus isolates ...

  4. Synthesis and processing in Escherichia coli of human leucocyte interferon fused with the signal sequence of Bacillus amyloliquefaciens a-amylase

    International Nuclear Information System (INIS)

    Sorokin, A.V.; Avakov, A.S.; Bogush, V.G.

    1985-01-01

    Earlier, the authors reported cloning of the alpha-amylase gene of B. amyloliquefaciens in B. subtilis and E. coli. Currently, the authors report results on the expression of the hybrid gene consisting of the DNA fragment coding for the leader part of B. amyloliquefaciens alpha-amylase and the structural part of the human interferon alpha-2 in E. coli cells. This gene contains an additional methionine codon at the 5'-terminal, which codes for the interferon structure (without its own signal peptide). The interferon gene was inserted into plasmid /sub p/TG 278 at the cleavage site of EcoRI. The structure of the plasmid thus obtained the signal peptide of amylase, five amino acids (Val-Gly-Glu-Phe-Met), and the structural part of the interferon. The E. coli C600 cells carrying plasmid pTGA6 were used to study interferon secretions. The interferon activity was determined radioimmunologically with the use of monoclonal anti-bodies NK2

  5. Investigation of biosurfactant production by Bacillus pumilus 1529 and Bacillus subtilis WPI

    Directory of Open Access Journals (Sweden)

    shila khajavi shojaei

    2016-06-01

    Full Text Available Introduction: Biosurfactants are unique amphipathic molecules with extensive application in removing organic and metal contaminants. The purpose of this study was to investigate production of biosurfactant and determine optimal conditions to produce biosurfactant by Bacillus pumilus 1529 and Bacillus subtilis WPI. Materials and methods: In this study, effect of carbon source, temperature and incubation time on biosurfactant production was evaluated. Hemolytic activity, emulsification activity, oil spreading, drop collapse, cell hydrophobicity and measurement of surface tension were used to detect biosurfactant production. Then, according to the results, the optimal conditions for biosurfactant production by and Bacillus subtilis WPI was determined. Results: In this study, both bacteria were able to produce biosurfactant at an acceptable level. Glucose, kerosene, sugarcane molasses and phenanthrene used as a sole carbon source and energy for the mentioned bacteria. Bacillus subtilis WPI produced maximum biosurfactant in the medium containing kerosene and reduced surface tension of the medium to 33.1 mN/m after 156 hours of the cultivation at 37°C. Also, the highest surface tension reduction by Bacillus pumilus 1529 occurred in the medium containing sugarcane molasses and reduce the surface tension of culture medium after 156 hours at 37°C from 50.4 to 28.83 mN/m. Discussion and conclusion: Bacillus pumilus 1529 and Bacillus subtilis WPI had high potential in production of biosurfactant and degradation of petroleum hydrocarbons and Phenanthrene. Therefore, it could be said that these bacteria had a great potential for applications in bioremediation and other environmental process.

  6. Current development in genetic engineering strategies of Bacillus species

    Science.gov (United States)

    2014-01-01

    The complete sequencing and annotation of the genomes of industrially-important Bacillus species has enhanced our understanding of their properties, and allowed advances in genetic manipulations in other Bacillus species. Post-genomic studies require simple and highly efficient tools to enable genetic manipulation. Here, we summarize the recent progress in genetic engineering strategies for Bacillus species. We review the available genetic tools that have been developed in Bacillus species, as well as methods developed in other species that may also be applicable in Bacillus. Furthermore, we address the limitations and challenges of the existing methods, and discuss the future research prospects in developing novel and useful tools for genetic modification of Bacillus species. PMID:24885003

  7. Modeling Radiation Effectiveness for Inactivation of Bacillus Spores

    Science.gov (United States)

    2015-09-17

    MODELING RADIATION EFFECTIVENESS FOR INACTIVATION OF BACILLUS SPORES DISSERTATION Emily A. Knight, Major, USAF AFIT-ENC-DS-15-S-001 DEPARTMENT OF THE...not subject to copyright protection in the United States. AFIT-ENC-DS-15-S-001 MODELING RADIATION EFFECTIVENESS FOR INACTIVATION OF BACILLUS SPORES...EFFECTIVENESS FOR INACTIVATION OF BACILLUS SPORES Emily A. Knight, B.A., M.S. Major, USAF Committee Membership: Dr. William P. Baker Chair Dr. Larry W

  8. Bacillus caldolyticus prs gene encoding phosphoribosyldiphosphate synthase

    DEFF Research Database (Denmark)

    Krath, Britta N.; Hove-Jensen, Bjarne

    1996-01-01

    The prs gene, encoding phosphoribosyl-diphosphate (PRPP) synthase, as well as the flanking DNA sequences were cloned and sequenced from the Gram-positive thermophile, Bacillus caldolyticus. Comparison with the homologous sequences from the mesophile, Bacillus subtilis, revealed a gene (gca......D) encoding N-acetylglucosamine-l-phosphate uridyltransferase upstream of prs, and a gene homologous to ctc downstream of prs. cDNA synthesis with a B. caldolyticus gcaD-prs-ctc-specified mRNA as template, followed by amplification utilising the polymerase chain reaction indicated that the three genes are co......-transcribed. Comparison of amino acid sequences revealed a high similarity among PRPP synthases across a wide phylogenetic range. An E. coli strain harbouring the B. caldolyticus prs gene in a multicopy plasmid produced PRPP synthase activity 33-fold over the activity of a haploid B. caldolyticus strain. B. caldolyticus...

  9. Bioakumulasi logam berat Cu oleh Bacillus sp

    Directory of Open Access Journals (Sweden)

    Riesta Primaharinastiti

    2012-02-01

    Full Text Available The research was conducted to investigate the ability of Bacillus sp in accumulating Cu and how much it can be acumulated. Themedium used to growth the bacterium was Nutrient Broth and Atomic Absorption Spectrophotometry methods was used to assay theCu, both in the cells and medium. The result of this study showed that Bacillus sp incubated in the Nutrient Broth medium containing10 ppm of Cu, with continuous stirring in the room temperature was able to reduce Cu in the medium 8.912–12.623% and accumulateCu in the cell 0.1149–0.1400 %/mg cells. Based on this result, it is necessary to develop more studies to find out what factors thatinfluence the accumulation process and to optimize the bioprocess.

  10. Regulation of glutamate dehydrogenase in Bacillus subtilis.

    OpenAIRE

    Kane, J F; Wakim, J; Fischer, R S

    1981-01-01

    The activity of the nicotinamide adenine dinucleotide-dependent glutamate dehydrogenase in Bacillus subtilis was influenced by the carbon source, but not the nitrogen source, in the growth medium. The highest specific activity for this enzyme was found when B. subtilis was grown in a minimal or rich medium that contained glutamate as the carbon source. It is proposed that glutamate dehydrogenase serves a catabolic function in the metabolism of glutamate, is induced by glutamate, and is subjec...

  11. Regulation of glutamate dehydrogenase in Bacillus subtilis.

    Science.gov (United States)

    Kane, J F; Wakim, J; Fischer, R S

    1981-01-01

    The activity of the nicotinamide adenine dinucleotide-dependent glutamate dehydrogenase in Bacillus subtilis was influenced by the carbon source, but not the nitrogen source, in the growth medium. The highest specific activity for this enzyme was found when B. subtilis was grown in a minimal or rich medium that contained glutamate as the carbon source. It is proposed that glutamate dehydrogenase serves a catabolic function in the metabolism of glutamate, is induced by glutamate, and is subject to catabolite repression. PMID:6118356

  12. Identification of Fatty Acids in Bacillus cereus.

    Science.gov (United States)

    Ginies, Christian; Brillard, Julien; Nguyen-The, Christophe

    2016-12-05

    The Bacillus species contain branched chain and unsaturated fatty acids (FAs) with diverse positions of the methyl branch (iso or anteiso) and of the double bond. Changes in FA composition play a crucial role in the adaptation of bacteria to their environment. These modifications entail a change in the ratio of iso versus anteiso branched FAs, and in the proportion of unsaturated FAs relative to saturated FAs, with double bonds created at specific positions. Precise identification of the FA profile is necessary to understand the adaptation mechanisms of Bacillus species. Many of the FAs from Bacillus are not commercially available. The strategy proposed herein identifies FAs by combining information on the retention time (by calculation of the equivalent chain length (ECL)) with the mass spectra of three types of FA derivatives: fatty acid methyl esters (FAMEs), 4,4-dimethyl oxazoline derivatives (DMOX), and 3-pyridylcarbinyl ester (picolinyl). This method can identify the FAs without the need to purify the unknown FAs. Comparing chromatographic profiles of FAME prepared from Bacillus cereus with a commercial mixture of standards allows for the identification of straight-chain saturated FAs, the calculation of the ECL, and hypotheses on the identity of the other FAs. FAMEs of branched saturated FAs, iso or anteiso, display a constant negative shift in the ECL, compared to linear saturated FAs with the same number of carbons. FAMEs of unsaturated FAs can be detected by the mass of their molecular ions, and result in a positive shift in the ECL compared to the corresponding saturated FAs. The branching position of FAs and the double bond position of unsaturated FAs can be identified by the electron ionization mass spectra of picolinyl and DMOX derivatives, respectively. This approach identifies all the unknown saturated branched FAs, unsaturated straight-chain FAs and unsaturated branched FAs from the B. cereus extract.

  13. Isolation of bacillus thuringiensis from different samples from Mansehra District

    International Nuclear Information System (INIS)

    Younis, F.; Lodhi, A.F.; Raza, G.

    2009-01-01

    The insecticidal activity of Bacillus thuringiensis has made it very interesting for the control of a variety of agricultural pests and human disease vectors. The present study is an attempt to explore the potential and diversity. of Bacillus thuringiensis. from the local environment for the control of cotton spotted bollworm (Earias sp.), a major pest of cotton. Two hundred and ninety eight samples of soil, grain dust, wild animal dung, birds dropping, decaying leaves and dead insects were collected from different ecological environments of Mansehra District yielding 438 Bacillus thuringiensis isolates that produce parasporal crystalline inclusions. In this study the soil samples were found to be the richest source for Bacillus thuringiensis. (author)

  14. Bacillus cellulasensis sp. nov., isolated from marine sediment.

    Science.gov (United States)

    Mawlankar, Rahul; Thorat, Meghana N; Krishnamurthi, Srinivasan; Dastager, Syed G

    2016-01-01

    A novel bacterial strain NIO-1130(T) was isolated from sediment sample taken from Chorao Island, Goa Province, India, and subjected to a taxonomic investigation. The strain was Gram-positive, aerobic, and motile. Phylogenetic analysis based on 16S rRNA gene sequences placed the isolate within the genus Bacillus and strain NIO-1130(T) showed highest sequence similarity with Bacillus halosaccharovorans DSM 25387(T) (98.4%) and Bacillus niabensis CIP 109816(T) (98.1%), whereas other Bacillus species showed bacillus group. The major menaquinone was MK-7 and the predominant cellular fatty acids were iso-C15:0, anteiso-C15:0, iso-C17:0, and anteiso-C17:0. The strain showed a DNA G+C content of 39.9 mol%. DNA-DNA hybridization studies revealed that strain NIO-1130(T) exhibits 70% similarity with Bacillus halosaccharovorans DSM 25387(T) and Bacillus niabensis CIP 109816(T). On the basis of physiological, biochemical, chemotaxonomic and phylogenetic analyses, we consider the isolate to represent a novel species of the genus Bacillus, for which the name Bacillus cellulasensis sp. nov., is proposed. The type strain is NIO-1130(T) (=NCIM 5461(T)=CCTCC AB 2011126(T)).

  15. Complete Genomes of Bacillus coagulans S-lac and Bacillus subtilis TO-A JPC, Two Phylogenetically Distinct Probiotics

    Science.gov (United States)

    Ramya, T. N. C.; Subramanian, Srikrishna

    2016-01-01

    Several spore-forming strains of Bacillus are marketed as probiotics due to their ability to survive harsh gastrointestinal conditions and confer health benefits to the host. We report the complete genomes of two commercially available probiotics, Bacillus coagulans S-lac and Bacillus subtilis TO-A JPC, and compare them with the genomes of other Bacillus and Lactobacillus. The taxonomic position of both organisms was established with a maximum-likelihood tree based on twenty six housekeeping proteins. Analysis of all probiotic strains of Bacillus and Lactobacillus reveal that the essential sporulation proteins are conserved in all Bacillus probiotic strains while they are absent in Lactobacillus spp. We identified various antibiotic resistance, stress-related, and adhesion-related domains in these organisms, which likely provide support in exerting probiotic action by enabling adhesion to host epithelial cells and survival during antibiotic treatment and harsh conditions. PMID:27258038

  16. Complete Genomes of Bacillus coagulans S-lac and Bacillus subtilis TO-A JPC, Two Phylogenetically Distinct Probiotics.

    Directory of Open Access Journals (Sweden)

    Indu Khatri

    Full Text Available Several spore-forming strains of Bacillus are marketed as probiotics due to their ability to survive harsh gastrointestinal conditions and confer health benefits to the host. We report the complete genomes of two commercially available probiotics, Bacillus coagulans S-lac and Bacillus subtilis TO-A JPC, and compare them with the genomes of other Bacillus and Lactobacillus. The taxonomic position of both organisms was established with a maximum-likelihood tree based on twenty six housekeeping proteins. Analysis of all probiotic strains of Bacillus and Lactobacillus reveal that the essential sporulation proteins are conserved in all Bacillus probiotic strains while they are absent in Lactobacillus spp. We identified various antibiotic resistance, stress-related, and adhesion-related domains in these organisms, which likely provide support in exerting probiotic action by enabling adhesion to host epithelial cells and survival during antibiotic treatment and harsh conditions.

  17. Complete Genomes of Bacillus coagulans S-lac and Bacillus subtilis TO-A JPC, Two Phylogenetically Distinct Probiotics.

    Science.gov (United States)

    Khatri, Indu; Sharma, Shailza; Ramya, T N C; Subramanian, Srikrishna

    2016-01-01

    Several spore-forming strains of Bacillus are marketed as probiotics due to their ability to survive harsh gastrointestinal conditions and confer health benefits to the host. We report the complete genomes of two commercially available probiotics, Bacillus coagulans S-lac and Bacillus subtilis TO-A JPC, and compare them with the genomes of other Bacillus and Lactobacillus. The taxonomic position of both organisms was established with a maximum-likelihood tree based on twenty six housekeeping proteins. Analysis of all probiotic strains of Bacillus and Lactobacillus reveal that the essential sporulation proteins are conserved in all Bacillus probiotic strains while they are absent in Lactobacillus spp. We identified various antibiotic resistance, stress-related, and adhesion-related domains in these organisms, which likely provide support in exerting probiotic action by enabling adhesion to host epithelial cells and survival during antibiotic treatment and harsh conditions.

  18. Bioaccumulation of copper, zinc, cadmium and lead by Bacillus sp., Bacillus cereus, Bacillus sphaericus and Bacillus subtilis Bioacumulação de cobre, zinco, cádmio e chumbo por Bacillus sp., Bacillus cereus, Bacillus sphaericus e Bacillus subtilis

    Directory of Open Access Journals (Sweden)

    Antonio Carlos Augusto da Costa

    2001-03-01

    Full Text Available This work presents some results on the use of microbes from the genus Bacillus for uptake of cadmium, zinc, copper and lead ions. Maximum copper bioaccumulations were 5.6 mol/g biomass for B. sphaericus, 5.9 mol/g biomass for B. cereus and B. subtilis, and 6.4 mol/g biomass for Bacillus sp. Maximum zinc bioaccumulations were 4.3 mol/g biomass for B. sphaericus, 4.6 mol/g biomass for B. cereus, 4.8 mol/g biomass for Bacillus sp. and 5.0 mol/g biomass for B. subtilis. Maximum cadmium bioaccumulations were 8.0 mol/g biomass for B. cereus, 9.5 mol/g biomass for B. subtilis, 10.8 mol/g biomass for Bacillus sp. and 11.8 mol/g biomass for B. sphaericus. Maximum lead biomaccumulations were 0.7 mol/g biomass for B. sphaericus, 1.1 mol/g biomass for B. cereus, 1.4 mol/g biomass for Bacillus sp. and 1.8 mol/g biomass for B. subtilis. The different Bacillus strains tested presented distinct uptake capacities, and the best results were obtained for B. subtilis and B. cereus.Este trabalho apresenta resultados de acumulação dos íons metálicos cádmio, zinco, cobre e chumbo por bactérias do gênero Bacillus. A bioacumulação máxima de cobre foi 5,6 mol/g biomassa para B. sphaericus, 5,9 mol/g biomassa para B. cereus e B. subtilis, e 6,4 mol/g biomassa para Bacillus sp.. A bioacumulação máxima de zinco foi 4,3 mol/g biomassa para B. sphaericus, 4,6 mol/g biomassa para B. cereus, 4,8 mol/g biomassa para Bacillus sp. e 5,0 mol/g biomassa para B. subtilis. A bioacumulação máxima de cádmio foi 8,0 mol/g biomassa para B. cereus, 9,5 mol/g biomassa para B. subtilis, 10,8 mol/g biomassa para Bacillus sp. e 11,8 mol/g biomassa para B. sphaericus. A bioacumulação máxima de chumbo foi 0,7 mol/g biomassa para B. sphaericus, 1,1 mol/g biomassa para B. cereus, 1,4 mol/g biomassa para Bacillus sp. e 1,8 mol/g biomassa para B. subtilis. As distintas linhagens de Bacillus testadas apresentaram variáveis capacidades de carregamento de íons metálicos, sendo os

  19. Eucalyptus growth promotion by endophytic Bacillus spp.

    Science.gov (United States)

    Paz, I C P; Santin, R C M; Guimarães, A M; Rosa, O P P; Dias, A C F; Quecine, M C; Azevedo, J L; Matsumura, A T S

    2012-10-11

    Clonal eucalyptus plantings have increased in recent years; however, some clones with high production characteristics have vegetative propagation problems because of weak root and aerial development. Endophytic microorganisms live inside healthy plants without causing any damage to their hosts and can be beneficial, acting as plant growth promoters. We isolated endophytic bacteria from eucalyptus plants and evaluated their potential in plant growth promotion of clonal plantlets of Eucalyptus urophylla x E. grandis, known as the hybrid, E. urograndis. Eighteen isolates of E. urograndis, clone 4622, were tested for plant growth promotion using the same clone. These isolates were also evaluated for indole acetic acid production and their potential for nitrogen fixation and phosphate solubilization. The isolates were identified by partial sequencing of 16S rRNA. Bacillus subtilis was the most prevalent species. Several Bacillus species, including B. licheniformis and B. subtilis, were found for the first time as endophytes of eucalyptus. Bacillus sp strain EUCB 10 significantly increased the growth of the root and aerial parts of eucalyptus plantlets under greenhouse conditions, during the summer and winter seasons.

  20. Occurrence and significance of Bacillus cereus and Bacillus thuringiensis in ready-to-eat food

    DEFF Research Database (Denmark)

    Rosenquist, Hanne; Ørum-Smidt, Lasse; Andersen, Sigrid R

    2005-01-01

    Among 48,901 samples of ready-to-eat food products at the Danish retail market, 0.5% had counts of Bacillus cereus-like bacteria above 10(4) cfu g(-1). The high counts were most frequently found in starchy, cooked products, but also in fresh cucumbers and tomatoes. Forty randomly selected strains...... had at least one gene or component involved in human diarrhoeal disease, while emetic toxin was related to only one B. cereus strain. A new observation was that 31 out of the 40 randomly selected B. cereus-like strains could be classified as Bacillus thuringiensis due to crystal production and...

  1. Otimização das condições de cultivo para a produção de amilases pelo termofílico Bacillus sp. e hidrólise de amidos pela ação da enzima Optimization of culture conditions for the production of amylases by thermophilic Bacillus sp. and hydrolysis of starches by the action of the enzymes

    Directory of Open Access Journals (Sweden)

    Raquel Vieira de Carvalho

    2008-06-01

    quantidades de açúcares redutores.The optimization of culture conditions for the production of α-amylase by the thermophilic Bacillus sp strain SMIA-2 was carried out. In addition, the enzymatic hydrolysis of starch from several sources, such as potato, cassava and corn was investigated. Alpha-amylase production by Bacillus sp SMIA-2 cultivated in liquid cultures containing starch as carbon source and supplemented with 0.5 g.L-1 whey protein and 2.0 g.L-1 peptone reached a maximum of 37 U.mL-1 at 32 hours. The microorganism was capable of utilizing several carbon sources, but amylase activity varied with each source. Starch was the best carbon source for amylase secretion, while lactose, maltose, sucrose, galactose, and glucose were not very effective. Decreasing starch concentration in the medium to 2.5 g.L-1 improved organism growth and enzyme activity. At higher starch concentrations, enzyme production was comparatively lower. Among the various organic and inorganic nitrogen sources, peptone (2.0 g.L-1 was found to be the best. Regarding the amount of whey protein in the medium, the concentration of 0.25 g.L-1 was considered the most effective for amylase secretion by the organism. Maximum amylase activity was observed at 50 °C and pH 8.5. The enzyme was able to degrade all the starches tested. Potato starch hydrolysis resulted in a higher yield of reducing sugars in comparison to the other starches. Soluble and cassava starch were, respectively, in second and third positions regarding the liberation of reducing sugars, while the amylase studied showed a slightly lower affinity for corn starch. Increasing the reaction temperature to 70 °C resulted in higher levels of reducing sugars after the hydrolysis of the substrates, except for soluble starch.

  2. Biodiversity of predominant Bacillus isolated from afitin, iru and ...

    African Journals Online (AJOL)

    Two hundred (200) presumptive isolates of Bacillus collected at different fermentation time from spontaneous fermented samples of afitin, iru and sonru produced in three different regions of Benin were identified at species and strains levels. ITS-PCR-RFLP revealed that 79% of the isolates were really identified as Bacillus, ...

  3. The use of two biological formulations of Bacillus Thuringiensis and ...

    African Journals Online (AJOL)

    Two biological formulations of the microbial agents Bacillus thuringiensis and Bacillus sphaericus, known by their trade names Vectobac 12 AS and VectoLex CG (Corn Cob) granules, respectively, were obtained from Valent Biosciences Company (formerly Abbott Laboratories) of North Chicago, USA, and applied to control ...

  4. Production of cellulase by a novel cellulolytic Bacillus sp

    African Journals Online (AJOL)

    squ

    2016-10-26

    Oct 26, 2016 ... Eight cellulose degrading bacteria were isolated from compost and were identified as Bacillus licheniformis by 16S rRNA sequencing. Among the eight isolates, Bacillus licheniformis B4, B7 and B8 showed the highest cellulase activity. B. licheniformis B4 and B8 showed the maximum cellulase.

  5. Isolation and characterization of a novel Bacillus subtilis WD23 ...

    African Journals Online (AJOL)

    The strain Bacillus sp. WD23 exhibiting laccase activity was screened from forest soil. The M9 medium containing Cu2+ was used for enriching and isolating bacterial strains capable of oxidizing syringaldazine. One isolated strain was identified as Bacillus subtilis WD23 based on the results of physiological and biochemical ...

  6. Monitoring of marine Bacillus diversity among the bacteria ...

    African Journals Online (AJOL)

    germination an indication of manganese reduction by Bacillus. Two strains identified as Bacillus pumilus using 16S rRNA gene sequence were isolated from Buzzers Bay seawater at 45 feet dept. The strains spores were centrally located and they were able to tolerate 1.0 M NaCl concentration indicating their marine origin.

  7. Biodegradation of naphthalene and phenanthren by Bacillus subtilis 3KP

    Science.gov (United States)

    Ni'matuzahroh, Trikurniadewi, N.; Pramadita, A. R. A.; Pratiwi, I. A.; Salamun, Fatimah, Sumarsih, Sri

    2017-06-01

    The purposes of this research were to know growth response, degradation ability, and uptake mechanism of naphthalene and phenanthrene by Bacillus subtilis 3KP. Bacillus subtilis 3KP was grown on Mineral Synthetic (MS) medium with addition of 1% yeast extract and naphthalene and phenanthrene respectively 200 ppm in different cultures. Bacillus subtilis 3KP growth response was monitored by Total Plate Count (TPC) method, the degradation ability was monitored by UV-Vis spectrophotometer, and the uptake mechanism of hydrocarbon was monitored by emulsification activity, decrease of surface tension, and activity of Bacterial Adherence to Hydrocarbon (BATH). Bacillus subtilis 3KP was able to grow and show biphasic growth pattern on both of substrates. Naphthalene and phenanthrene were used as a carbon source for Bacillus subtilis 3KP growth that indicated by the reduction of substrate concomitant with the growth. At room temperature conditions (± 30°C) and 90 rpm of agitation for 7 days, Bacillus subtilis 3KP could degrade naphthalene in the amount of 70.5% and phenanthrene in the amount of 24.8%. Based on the analysis of UV-Vis spectrophotometer, three metabolites, 1-hydroxy-2-naphthoic acid, salicylic acid, and pyrocatechol were found in both cultures. The metabolite identification became basis of propose degradation pathway of naphthalene and phenanthrene by Bacillus subtilis 3KP. The results of hydrocarbon uptake mechanism test show that Bacillus subtilis 3KP used all of the mechanism to degrade naphthalene and phenanthrene.

  8. Isolation and characterization of Bacillus thuringiensis from soils in ...

    African Journals Online (AJOL)

    Of 110 Bacillus thuringiensis isolates analyzed for the presence of crystal protein genes, 7 tested positive for cry 4, cry 11, and cyt toxin genes. Sequencing of these genes in positive strains demonstrated 99–100 % homology to known mosquitocidal cry and cyt genes in Bacillus thuringiensis subsp. israelensis. The present ...

  9. Hydrolytic and synthetic activities of esterases produced by Bacillus ...

    African Journals Online (AJOL)

    A novel esterase producer strain named Bacillus sp. A60 was isolated from a soil sample contaminated with hydrocarbons. It was found to belong to Bacillus subtilis species through morphological, biochemical and 16S rRNA gene sequence analyses. This strain which can tolerate 15% (w/v) NaCl and growth at 55°C, ...

  10. by lipase from Bacillus thuringiensis and Lysinibacillus sphaericus

    African Journals Online (AJOL)

    This study reported production of lipase by Bacillus thuringiensis and Lysinibacillus sphaericus. Bacteria isolates were screened on Bushnell-Hass Mineral Salt medium containing 1% PMS for oil degradation. Two potent isolates were identified using 16S rRNA as Bacillus thuringiensis and Lysinibacillus sphaericus.

  11. Role of fatty acids in Bacillus environmental adaptation

    Directory of Open Access Journals (Sweden)

    Sara Esther Diomande

    2015-08-01

    Full Text Available The large bacterial genus genus Bacillus is widely distributed in the environment and is able to colonize highly diverse niches. Some Bacillus species harbour pathogenic characteristics. The fatty acid (FA composition is among the essential criteria used to define Bacillus species. Some elements of the FA pattern composition are common to Bacillus species, whereas others are specific and can be categorized in relation to the ecological niches of the species. Bacillus species are able to modify their FA patterns to adapt to a wide range of environmental changes, including changes in the growth medium, temperature, food processing conditions, and pH. Like many other Gram-positive bacteria, Bacillus strains display a well-defined FA synthesis II system that is equilibrated with a FA degradation pathway and regulated to efficiently respond to the needs of the cell. Like endogenous FAs, exogenous FAs may positively or negatively affect the survival of Bacillus vegetative cells and the spore germination ability in a given environment. Some of these exogenous FAs may provide a powerful strategy for preserving food against contamination by the Bacillus pathogenic strains responsible for foodborne illness.

  12. Evaluation of the Larvicidal Activities of Bacillus Sphaericus on ...

    African Journals Online (AJOL)

    Evaluation of the Larvicidal Activities of Bacillus Sphaericus on Culex Mosquito Found in Sokoto. SB Manga, M Galadima. Abstract. Studies on the larvicidal activities of Bacillus sphaericus, against Culex quinquefasciatus mosquitoes found in this area (Sokoto), was carried out. The B. sphaericus (SPH 88) was obtained ...

  13. Evaluation of antifungal activity from Bacillus strains against ...

    African Journals Online (AJOL)

    In this study, 30 bacterial strains isolated from marine biofilms were screened for their antifungal activity against Rhizoctonia solani by dual culture assay. Two bacterial strains, Bacillus subtilis and Bacillus cereus, showed a clear antagonism against R. solani on potato dextrose agar (PDA) medium. The antagonistic activity ...

  14. Antagonistic activity of selected strains of Bacillus thuringiensis ...

    African Journals Online (AJOL)

    The aim of this work was to determine, in vitro, the antagonistic effectiveness of 60 strains of Bacillus thuringiensis against damping-off and root and stem rot caused by Rhizoctonia solani. The strains were obtained from the International Collection of Entomopathogenic Bacillus at the FCB-UANL. During the in vitro dual ...

  15. Inactivation of Bacillus Anthracis Spores Using Carbon Nanotubes

    Science.gov (United States)

    2014-10-30

    effect of SWNTs in combination with antimicrobial chemicals on inactivation of B. anthracis spores; 4) the effect of CNTs coated surfaces on the...2010 31-May-2014 Approved for Public Release; Distribution Unlimited Final Report: (Life Science Division/ Biochemistry ) Inactivation of Bacillus... Biochemistry ) Inactivation of Bacillus Anthracis Spores Using Carbon Nanotubes Report Title The Specific Aims of the project were to investigate: 1) the

  16. Recombinant EXLX1 from Bacillus subtilis for enhancing enzymatic ...

    African Journals Online (AJOL)

    Recombinant EXLX1 from Bacillus subtilis for enhancing enzymatic hydrolysis of corn stover with low cellulase loadings. ... These results provided a feasible way for the potential application of BsEXLX1 in the efficient saccharification of cellulose materials for bioethanol production. Key word: Bacillus subtilis, BsEXLX1, ...

  17. Genome analysis-based reclassification of Bacillus weihenstephanensis as a later heterotypic synonym of Bacillus mycoides.

    Science.gov (United States)

    Liu, Yang; Lai, Qiliang; Shao, Zongze

    2018-01-01

    The aim of this study was to clarify the taxonomic status of the species Bacillus weihenstephanensis. A complete genome sequence for the type strain of B. weihenstephanensis was compared against that of the closely related type strain of Bacillus mycoides. The digital DNA-DNA hybridization and average nucleotide identity values between the two type strains was greater than two recognized thresholds for bacterial species delineation, indicating that they should belong to the same genomospecies. The psychrotolerant characteristic and signature sequences of 16S rRNA and cspA genes were incapable of distinguishing B. weihenstephanensis from some non-B. weihenstephanensis strains. Meanwhile, the metabolic, physiological and chemotaxonomic features for the type strain of B. weihenstephanensis were shown to be congruent with those of B. mycoides. On this basis, the taxonomic affiliations of related strains from the Genbank database were determined using multilocus sequence typing and genomic analyses. Therefore, we propose Bacillus weihenstephanensis as a later heterotypic synonym of Bacillus mycoides and correction of erroneous species identifications for several strains.

  18. Identification and characterization of clinical Bacillus spp. isolates phenotypically similar to Bacillus anthracis.

    Science.gov (United States)

    Beesley, Cari A; Vanner, Cynthia L; Helsel, Leta O; Gee, Jay E; Hoffmaster, Alex R

    2010-12-01

    Bacillus anthracis, the etiological agent of anthrax, is a gram-positive, spore-forming rod, with colonies exhibiting a unique ground-glass appearance, and lacking hemolysis and motility. In addition to these phenotypes, several others traits are characteristic of B. anthracis such as susceptibility to gamma phage, the presence of two virulence plasmids (pX01 and pX02), and specific cell wall and capsular antigens that are commonly detected by direct fluorescent-antibody assays. We report on the identification and characterization of 14 Bacillus megaterium and four Bacillus sp. clinical isolates that are nonhemolytic, nonmotile, and produce a capsule antigenically similar to B. anthracis. This work furthers our understanding of Bacillus diversity and the limitations of the assays and phenotypes that are used to differentiate species in this genus. Further work is necessary to understand whether these strains are opportunistic pathogens or just contaminates. FEMS Microbiology Letters © 2010 Federation of European Microbiological Societies. Published by Blackwell Publishing Ltd. No claim to original US government works.

  19. Bacillus swezeyi sp. nov. and Bacillus haynesii sp. nov., isolated from desert soil.

    Science.gov (United States)

    Dunlap, Christopher A; Schisler, David A; Perry, Elizabeth B; Connor, Nora; Cohan, Frederick M; Rooney, Alejandro P

    2017-08-01

    Two isolates of Gram-reaction-positive, facultatively anaerobic, motile, rod-shaped, endospore-forming bacteria were identified during a survey of the diversity of strains belonging to the genus Bacillus deposited in the Agriculture Research Service Culture Collection. These strains were originally isolated from soil in Evolution Canyon III (Israel) in a survey of ecological diversification. Phylogenetic analysis of the 16S rRNA gene of strains NRRL B-41294T and NRRL B-41327T determined they were closely related to members of the Bacillus licheniformis clade. The genome of each strain was sequenced, and further analysis indicated that the strains represented unique species based on in silico DNA-DNA hybridization analyses. A phylogenomic analysis revealed that NRRL B-41294T and NRRL B-41327T were closely related to the group that includes B. licheniformis. In phenotypic characterization, both NRRL B-41294T and NRRL B-41327T were found to grow at temperatures of between 15 and 60 °C and tolerated up to 12 % NaCl (w/v). The predominant cellular fatty acids were anteiso-C15 : 0 and iso-C15 : 0, and peptidoglycan from cell walls contained meso-diaminopimelic acid. The DNA G+C content was 45.7 and 44.3 mol% for NRRL B-41327T and NRRL B-41294T, respectively. Furthermore, each strain had a unique carbon utilization pattern that distinguished it from its nearest phylogenetic neighbours. Based upon the consensus of phylogenetic and phenotypic analyses, we conclude that these strains represent two novel species within the genus Bacillus, for which the name Bacillus swezeyi sp. nov. is proposed, with type strain NRRL B-41294T (=CCUG 70177T), and the name Bacillus haynesii sp. nov. is proposed, with type strain NRRL B-41327T (=CCUG 70178T).

  20. Emetic food poisoning caused by Bacillus cereus.

    Science.gov (United States)

    Holmes, J R; Plunkett, T; Pate, P; Roper, W L; Alexander, W J

    1981-05-01

    Symptoms of acute food poisoning developed in eight members of a group who ate lunch at a cafeteria. After brief incubation periods, all affected individuals complained of nausea and abdominal cramps. Four persons promptly experienced vomiting. None of those affected was found to have fever and all recovered with 48 hours. Epidemiologic investigation incriminated macaroni and cheese as a cause of the illness and samples of this food contained large numbers of Bacillus cereus. Previous outbreaks of B cereus emetic food poisoning have been associated with consumption of contaminated fried rice and may occur after ingestion of other foods.

  1. Potential of Bacillus spp produces siderophores insuppressing thewilt disease of banana plants

    Science.gov (United States)

    Kesaulya, H.; Hasinu, J. V.; Tuhumury, G. NC

    2018-01-01

    In nature, different types of siderophore such as hydroxymate, catecholets and carboxylate, are produced by different bacteria. Bacillus spp were isolated from potato rhizospheric soil can produce siderophore of both catecholets and salicylate type with different concentrations. Various strains of Bacillus spp were tested for pathogen inhibition capability in a dual culture manner. The test results showed the ability of inhibition of pathogen isolated from banana wilt disease. From the result tested were found Bacillus niabensis Strain PT-32-1, Bacillus subtilis Strain SWI16b, Bacillus subtilis Strain HPC21, Bacillus mojavensis Strain JCEN3, and Bacillus subtilis Strain HPC24 showed different capabilities in suppressing pathogen.

  2. ABILITY OF BACTERIAL CONSORTIUM: Bacillus coagulans, Bacilus licheniformis, Bacillus pumilus, Bacillus subtilis, Nitrosomonas sp. and Pseudomonas putida IN BIOREMEDIATION OF WASTE WATER IN CISIRUNG WASTE WATER TREATMENT PLANT

    Directory of Open Access Journals (Sweden)

    Ratu SAFITRI

    2015-10-01

    Full Text Available This study was conducted in order to determine the ability of bacterial consortium: Bacillus coagulans, Bacilus licheniformis, Bacillus pumilus, Bacillus subtilis, Nitrosomonas sp., and Pseudomonas putida in bioremediation of wastewater origin Cisirung WWTP. This study uses an experimental method completely randomized design (CRD, which consists of two treatment factors (8x8 factorial design. The first factor is a consortium of bacteria (K, consisting of 8 level factors (k1, k2, k3, k4, k5, k6, k7, and k8. The second factor is the time (T, consisting of a 7 level factors (t0, t1, t2, t3, t4, t5, t6, and t7. Test parameters consist of BOD (Biochemical Oxygen Demand, COD (Chemical Oxygen Demand, TSS (Total Suspended Solid, Ammonia and Population of Microbes during bioremediation. Data were analyzed by ANOVA, followed by Duncan test. The results of this study showed that the consortium of Bacillus pumilus, Bacillus subtilis, Bacillus coagulans, Nitrosomonas sp., and Pseudomonas putida with inoculum concentration of 5% (k6 is a consortium of the most effective in reducing BOD 71.93%, 64.30% COD, TSS 94.85%, and 88.58% of ammonia.

  3. Karakterisasi protease Bacillus sp. UGM5

    Directory of Open Access Journals (Sweden)

    Titik Purwati Widowati

    1999-07-01

    Full Text Available The objective of this experiment is to indentify the characters of proetease produced by Bacillus sp.UGM5.the protease secreted by Bacillus sp.UGM5 was first isolated,purified and then charactirezed.The crude enzyme has spesific actifity of 1.14 U/mg,however,the spesific activity of purified enzyme was increased by 23.8 times fold and recovery was 33.69%.The Page of nondenatured crude enzymes showes two type of proreases,however ,the SDS-Page of denatured purified enzyme showed four protein-bends with molecular weights of 55.5 kDa,18kDa respecetively.The optimum pH and temperature for the enzyme acrivity are 8.5 and 420C and belongs to serin protease type,with Km 3 X 10-3mM and Vmax 0.0890mM/30 minutes.The activity is not inhibited by Ca+2,Fe+2 and EDTA.

  4. Identification of Bacillus anthracis by Using Monoclonal Antibody to Cell Wall Galactose-N-Acetylglucosamine Polysaccharide

    Science.gov (United States)

    1990-02-01

    Bacillus circulans ATCC 4513 b - - NR NT NT NT NT Bacillus coagulans ATCC 7050 b - - NR NT NT NT NT Bacillus eugilitis B-61 f - - NR NT NT NT NT...American Society for Microbiology W Identification of Bacillus anthracis by-U-sing Monoclonal Antibody CC to Cell Wall Galactose-N-Acetylglucosamine...Received 22 June 1989/Accepted 31 October 1989 ’ Guanidine extracts of crude Bacillus anthracis cell wall were used to vaccinate BALB/c mice and to

  5. Antifungal activity of indigenous Bacillus spp. isolated from soil

    Directory of Open Access Journals (Sweden)

    Bjelić Dragana Đ.

    2017-01-01

    Full Text Available Biocontrol using plant growth-promoting rhizobacteria (PGPR represents an alternative approach to disease management, since PGPR are known to promote growth and reduce diseases in various crops. Among the different PGPR, members of the genus Bacillus are prefered for most biotechnological uses due to their capability to form extremely resistant spores and produce a wide variety of metabolites with antimicrobial activity. The objective of this research was to identify antagonistic bacteria for management of the plant diseases. Eleven isolates of Bacillus spp. were obtained from the soil samples collected from different localities in the Province of Vojvodina. The antifungal activity of bacterial isolates against five fungal species was examined using a dual plate assay. Bacillus isolates exhibited the highest antifungal activity against Fusarium proliferatum, Fusarium oxysporum f. sp. cepae and Alternaria padwickii, while they had the least antagonistic effect on Fusarium verticillioides and Fusarium graminearum. Molecular identification showed that effective bacterial isolates were identified as Bacillus safensis (B2, Bacillus pumilus (B3, B11, Bacillus subtilis (B5, B7 and Bacillus megaterium (B8, B9. The highest antagonistic activity was exhibited by isolates B5 (from 39% to 62% reduction in fungal growth and B7 (from 40% to 71% reduction in fungal growth. These isolates of B. subtilis could be used as potential biocontrol agents of plant diseases. [Project of the Serbian Ministry of Education, Science and Technological Development, Grant no. TR-31073

  6. Heavy metals and their radionuclides uptake by Bacillus Licheniformis

    International Nuclear Information System (INIS)

    Ramadan, A.A.; Ahmed, M.M.; Abo-state, M.A.M.; Sarhan, M.; Faroqe, M.

    2007-01-01

    Bacillus licheniformis is a gram positive spore forming bacterium. Different concentrations of cobalt affected the ability of Co uptake and growth of Bacillus licheniformis. As the concentration increased, both the uptake and growth were decreased. Maximum Co uptake was found at ph 7.0, while for growth was ph 8.0. The optimum temperature for uptake and growth was 40 degree C and 20% inoculum size represents the maximum cobalt uptake by Bacillus licheniformis. Also, maximum uptake was recorded after 72 hours, incubation period. As the concentration of cesium was increased till 400 mg/l, the uptake was also increased. The optimum cesium uptake and growth was at ph 8.0. The optimum growth was at 45 degree C while Cs uptake was found at 35 degree C and 15% inoculum size represented the maximum Cs uptake. After 72 hour incubation period, maximum Cs uptake was recorded. Generally, Bacillus licheniformis removed more than 80% of Co and 50% of Cs from the broth medium. Addition of clay to Bacillus licheniformis increased both Co or Cs uptake. Bacillus licheniformis was gamma resistant and 10 KGy reduced the viability by 5.3 log cycles. The irradiated and non-irradiated cultures can grow on 500 or 700 mg Co or Cs. Bacillus licheniformis removed 99.32% of the Co radionuclides and 99.28% of Cs radionuclides

  7. Identification of Bacillus Strains for Biological Control of Catfish Pathogens

    Science.gov (United States)

    Ran, Chao; Carrias, Abel; Williams, Malachi A.; Capps, Nancy; Dan, Bui C. T.; Newton, Joseph C.; Kloepper, Joseph W.; Ooi, Ei L.; Browdy, Craig L.; Terhune, Jeffery S.; Liles, Mark R.

    2012-01-01

    Bacillus strains isolated from soil or channel catfish intestine were screened for their antagonism against Edwardsiella ictaluri and Aeromonas hydrophila, the causative agents of enteric septicemia of catfish (ESC) and motile aeromonad septicaemia (MAS), respectively. Twenty one strains were selected and their antagonistic activity against other aquatic pathogens was also tested. Each of the top 21 strains expressed antagonistic activity against multiple aquatic bacterial pathogens including Edwardsiella tarda, Streptococcus iniae, Yersinia ruckeri, Flavobacterium columnare, and/or the oomycete Saprolegnia ferax. Survival of the 21 Bacillus strains in the intestine of catfish was determined as Bacillus CFU/g of intestinal tissue of catfish after feeding Bacillus spore-supplemented feed for seven days followed by normal feed for three days. Five Bacillus strains that showed good antimicrobial activity and intestinal survival were incorporated into feed in spore form at a dose of 8×107 CFU/g and fed to channel catfish for 14 days before they were challenged by E. ictaluri in replicate. Two Bacillus subtilis strains conferred significant benefit in reducing catfish mortality (PBacillus strains also showed protective effects against E. ictaluri in striped catfish. Safety of the four strains exhibiting the strongest biological control in vivo was also investigated in terms of whether the strains contain plasmids or express resistance to clinically important antibiotics. The Bacillus strains identified from this study have good potential to mediate disease control as probiotic feed additives for catfish aquaculture. PMID:23029244

  8. Identification of Bacillus strains for biological control of catfish pathogens.

    Science.gov (United States)

    Ran, Chao; Carrias, Abel; Williams, Malachi A; Capps, Nancy; Dan, Bui C T; Newton, Joseph C; Kloepper, Joseph W; Ooi, Ei L; Browdy, Craig L; Terhune, Jeffery S; Liles, Mark R

    2012-01-01

    Bacillus strains isolated from soil or channel catfish intestine were screened for their antagonism against Edwardsiella ictaluri and Aeromonas hydrophila, the causative agents of enteric septicemia of catfish (ESC) and motile aeromonad septicaemia (MAS), respectively. Twenty one strains were selected and their antagonistic activity against other aquatic pathogens was also tested. Each of the top 21 strains expressed antagonistic activity against multiple aquatic bacterial pathogens including Edwardsiella tarda, Streptococcus iniae, Yersinia ruckeri, Flavobacterium columnare, and/or the oomycete Saprolegnia ferax. Survival of the 21 Bacillus strains in the intestine of catfish was determined as Bacillus CFU/g of intestinal tissue of catfish after feeding Bacillus spore-supplemented feed for seven days followed by normal feed for three days. Five Bacillus strains that showed good antimicrobial activity and intestinal survival were incorporated into feed in spore form at a dose of 8×10(7) CFU/g and fed to channel catfish for 14 days before they were challenged by E. ictaluri in replicate. Two Bacillus subtilis strains conferred significant benefit in reducing catfish mortality (PBacillus strains also showed protective effects against E. ictaluri in striped catfish. Safety of the four strains exhibiting the strongest biological control in vivo was also investigated in terms of whether the strains contain plasmids or express resistance to clinically important antibiotics. The Bacillus strains identified from this study have good potential to mediate disease control as probiotic feed additives for catfish aquaculture.

  9. Real-Time PCR Identification of Unique Bacillus anthracis Sequences.

    Science.gov (United States)

    Cieślik, P; Knap, J; Kolodziej, M; Mirski, T; Joniec, J; Graniak, G; Zakowska, D; Winnicka, I; Bielawska-Drózd, A

    2015-01-01

    Bacillus anthracis is a spore-forming, Gram-positive microorganism. It is a causative agent of anthrax, a highly infectious disease. It belongs to the "Bacillus cereus group", which contains other closely related species, including Bacillus cereus, Bacillus thuringiensis, Bacillus mycoides, Bacillus weihenstephanensis, and Bacillus pseudomycoides. B. anthracis naturally occurs in soil environments. The BA5345 genetic marker was used for highly specific detection of B. anthracis with TaqMan probes. The detection limit of a real-time PCR assay was estimated at the level of 16.9 copies (CI95% - 37.4 to 37.86, SD = 0.2; SE = 0.118). Oligonucleotides designed for the targeted sequences (within the tested locus) revealed 100 % homology to B. anthracis strain reference sequences deposited in the database (NCBI) and high specificity to all tested B. anthracis strains. Additional in silico analysis of plasmid markers pag and cap genes with B. anthracis strains included in the database was carried out. Our study clearly indicates that the BA5345 marker can be used with success as a chromosomal marker in routine identification of B. anthracis; moreover, detection of plasmid markers indicates virulence of the examined strains.

  10. Identification of strains Bacillus aerophilus MTCC 7304T as Bacillus altitudinis and Bacillus stratosphericus MTCC 7305T as a Proteus sp. and the status of the species Bacillus aeriusShivaji et al. 2006. Request for an Opinion.

    Science.gov (United States)

    Liu, Yang; Ramesh Kumar, N; Lai, Qiliang; Du, Juan; Dobritsa, Anatoly P; Samadpour, Mansour; Shao, Zongze

    2015-09-01

    On the basis of 16S rRNA, rpoB, gyrB and pycA gene sequence analyses, characterization of biochemical features and other phenotypic traits and pulsed-field gel electrophoresis (PFGE) fingerprinting, it was ascertained that strains Bacillus aerius MTCC 7303T, Bacillus aerophilus MTCC 7304(T) and Bacillus stratosphericus MTCC 7305(T) do not conform to the descriptions of the type strains of the respective species. Strains MTCC 7303(T) and MTCC 7304(T) were indistinguishable from Bacillus altitudinis DSM 21631(T), while strain MTCC 7305(T) should be classified as a representative of a Proteus sp. Our attempts to find other deposits of the type strains of these species were unsuccessful. Therefore, the results support the Request for an Opinion on the status of the species Bacillus aerophilus and Bacillus stratosphericus by Branquinho et al. [Branquinho, R., Klein, G., Kämpfer, P. & Peixe, L. V. (2015). Int J Syst Evol Microbiol 65, 1101]. It is also proposed that the Judicial Commission should place the name Bacillus aerius on the list of rejected names if a suitable replacement type strain cannot be found or a neotype is not proposed within two years following the publication of this Request (Rule 18c).

  11. Bacillus camelliae sp. nov., isolated from Pu'er tea.

    Science.gov (United States)

    Niu, Lili; Xiong, Mengjie; Zhang, Juan; Xiang, Yangquan; Song, Lei; Hua, Ziyi; Li, Wenying

    2018-02-01

    A novel aerobic, Gram-stain-positive, sporogenous, rod-shaped bacterial strain, 7578-1 T , was isolated from ripened Pu'er tea. Based on 16S rRNA gene sequence similarity comparisons, strain 7578-1 T was grouped into the genus Bacillus and appeared to be closely related to the type strains Bacillus shackletoniiLMG 18435 T (98.4 %), Bacillus acidicolaDSM 14745 T (97.6 %), Bacillus paralicheniformis KACC 18426 T (97.2 %) and Bacillus ginsengihumi KCTC 13944 T (96.7 %). The fatty acid profile containing the major fatty acids, iso-C15 : 0, anteiso-C15 : 0 and anteiso-C17 : 0 supported the allocation of strain 7578-1 T to the genus Bacillus. The strain had a cell-wall type A1γ peptidoglycan with meso-diaminopimelic acid as the diagnostic diamino acid. The major menaquinone was MK-7 (95 %). The predominant polar lipids were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylmethylethanolamine, phosphatidylethanolamine, one unidentified phospholipid and one unidentified lipid. The average nucleotide identity values between strain 7578-1 T and its most closely related species were 67.8-82.4 % by OrthoANIu analysis. The DNA-DNA relatedness value between strain 7578-1 T and the type strains of closely related species were 17-39 %, again indicating that strain 7578-1 T represented a novel species in the genus Bacillus. The DNA G+C content of strain 7578-1 T was 36.0 mol%. On the basis of the presented polyphasic evidence, strain 7578-1 T is considered to represent a novel species of the genus Bacillus, for which we propose the name Bacillus camelliae sp. nov. The type strain is 7578-1 T (=CGMCC 1.15374 T =KCTC 33845 T ).

  12. Architecture and Assembly of the Bacillus subtilis Spore Coat

    Science.gov (United States)

    2014-09-26

    483 489. 15. Abhyankar W, Ter Beek A, Dekker H, Kort R, Brul S, et al. (2011) Gel-free proteomic identification of the Bacillus subtilis insoluble coat... identification of additional sporulation genes in Bacillus subtilis. J Mol Biol 327: 945 972. AFM of Spore Coat Architecture PLOS ONE | www.plosone.org 16 September 2014 | Volume 9 | Issue 9 | e108560 ...1ITLE AND SUBTITLE 5a CONTRACTNUMBER Architecture and assembly of the Bacillus subtilis spore coat W911NF-09-l-0286 5b. GRANT NUMBER 5c. PROGRAM

  13. Multilocus sequence analysis of Bacillus thuringiensis serovars navarrensis, bolivia and vazensis and Bacillus weihenstephanensis reveals a common phylogeny.

    Science.gov (United States)

    Soufiane, Brahim; Baizet, Mathilde; Côté, Jean-Charles

    2013-01-01

    The Bacillus cereus group sensu lato includes six closely-related bacterial species: Bacillus cereus, Bacillus anthracis, Bacillus thuringiensis, Bacillus mycoides, Bacillus pseudomycoides and Bacillus weihenstephanensis. B. thuringiensis is distinguished from the other species mainly by the appearance of an inclusion body upon sporulation. B. weihenstephanensis is distinguished based on its psychrotolerance and the presence of specific signature sequences in the 16S rRNA gene and cspA genes. A total of seven housekeeping genes (glpF, gmK, ilvD, pta, purH, pycA and tpi) from different B. thuringiensis serovars and B. weihenstephanensis strains were amplified and their nucleotide sequences determined. A maximum likelihood phylogenetic tree was inferred from comparisons of the concatenated sequences. B. thuringiensis serovars navarrensis, bolivia and vazensis clustered not with the other B. thuringiensis serovars but rather with the B. weihenstephanensis strains, indicative of a common phylogeny. In addition, specific signature sequences and single nucleotide polymorphisms common to B. thuringiensis serovars navarrensis, bolivia and vazensis and the B. weihenstephanensis strains, and absent in the other B. thuringiensis serovars, were identified.

  14. Micro-Etched Platforms for Thermal Inactivation of Bacillus Anthracis and Bacillus Thuringiensis Spores

    Science.gov (United States)

    2008-03-01

    gentlemen are approachable and extremely well versed in their fields. Dr. Burggraf’s passion for learning coupled with the patience necessary to allow...wide spread employment on civilian targets has increased. Indeed, the well known biological agent Anthrax, Bacillus anthracis (B.a.), was recently...reached at 0.5 seconds with the heat from the fireball lasting a total of 4 seconds ( Orson , J.A. 2003; 104). From this data, the short time-temperature

  15. Description of Bacillus kexueae sp. nov. and Bacillus manusensis sp. nov., isolated from hydrothermal sediments.

    Science.gov (United States)

    Sun, Qing-Lei; Yu, Chao; Luan, Zhen-Dong; Lian, Chao; Hu, Yong-Hua; Sun, Li

    2018-03-01

    Two Gram-staining-positive, strictly aerobic bacilli, designated as strains Ma50-5 T and Ma50-6 T , were isolated from the hydrothermal sediments of Manus Basin in the western Pacific Ocean. Based on 16S rRNA gene sequence, strains Ma50-5 T and Ma50-6 T were most closely related to Bacillus alveayuensis (97.0 and 97.2 % identity, respectively). The 16S rRNA gene sequence identity between strains Ma50-5 T and Ma50-6 T was 97.4 %. The identities between strains Ma50-5 T and Ma50-6 T and other closely related organisms were below 97.0 %. The G+C contents of the genomic DNA of strains Ma50-5 T and Ma50-6 T were 43.4 and 47.6 mol%, respectively. The major fatty acids (>10 %) of both strains were iso-C15 : 0 and iso-C17 : 0. The predominant isoprenoid quinone detected in both strains was menaquinone-7. Phylogenetic, physiological, biochemical and morphological analyses suggested that strains Ma50-5 T and Ma50-6 T represent two novel species of the genus Bacillus, for which the names Bacillus kexueae sp. nov. (type strain Ma50-5 T =KCTC 33881 T =CCTCC AB 2017020 T ) and Bacillus manusensis sp. nov. (type strain Ma50-6 T =KCTC 33882 T =CCTCC AB 2017019 T ), respectively, are proposed.

  16. Characteristics and Application of a Novel Species of Bacillus: Bacillus velezensis.

    Science.gov (United States)

    Ye, Miao; Tang, Xiangfang; Yang, Ru; Zhang, Hongfu; Li, Fangshu; Tao, Fangzheng; Li, Fei; Wang, Zaigui

    2018-03-16

    Bacillus velezensis has been investigated and applied more and more widely recently because it can inhibit fungi and bacteria and become a potential biocontrol agent. In order to provide more clear and comprehensive understanding of B. velezensis for researchers, we collected the recent relevant articles systematically and reviewed the discovery and taxonomy, secondary metabolites, characteristics and application, gene function, and molecular research of B. velezensis. This review will give some direction to the research and application of this strain for the future.

  17. Genome sequence of Bacillus cereus and comparative analysis with Bacillus anthracis.

    Science.gov (United States)

    Ivanova, Natalia; Sorokin, Alexei; Anderson, Iain; Galleron, Nathalie; Candelon, Benjamin; Kapatral, Vinayak; Bhattacharyya, Anamitra; Reznik, Gary; Mikhailova, Natalia; Lapidus, Alla; Chu, Lien; Mazur, Michael; Goltsman, Eugene; Larsen, Niels; D'Souza, Mark; Walunas, Theresa; Grechkin, Yuri; Pusch, Gordon; Haselkorn, Robert; Fonstein, Michael; Ehrlich, S Dusko; Overbeek, Ross; Kyrpides, Nikos

    2003-05-01

    Bacillus cereus is an opportunistic pathogen causing food poisoning manifested by diarrhoeal or emetic syndromes. It is closely related to the animal and human pathogen Bacillus anthracis and the insect pathogen Bacillus thuringiensis, the former being used as a biological weapon and the latter as a pesticide. B. anthracis and B. thuringiensis are readily distinguished from B. cereus by the presence of plasmid-borne specific toxins (B. anthracis and B. thuringiensis) and capsule (B. anthracis). But phylogenetic studies based on the analysis of chromosomal genes bring controversial results, and it is unclear whether B. cereus, B. anthracis and B. thuringiensis are varieties of the same species or different species. Here we report the sequencing and analysis of the type strain B. cereus ATCC 14579. The complete genome sequence of B. cereus ATCC 14579 together with the gapped genome of B. anthracis A2012 enables us to perform comparative analysis, and hence to identify the genes that are conserved between B. cereus and B. anthracis, and the genes that are unique for each species. We use the former to clarify the phylogeny of the cereus group, and the latter to determine plasmid-independent species-specific markers.

  18. Construction of novel shuttle expression vectors for gene expression in Bacillus subtilis and Bacillus pumilus.

    Science.gov (United States)

    Shao, Huanhuan; Cao, Qinghua; Zhao, Hongyan; Tan, Xuemei; Feng, Hong

    2015-01-01

    A native plasmid (pSU01) was detected by genome sequencing of Bacillus subtilis strain S1-4. Two pSU01-based shuttle expression vectors pSU02-AP and pSU03-AP were constructed enabling stable replication in B. subtilis WB600. These vectors contained the reporter gene aprE, encoding an alkaline protease from Bacillus pumilus BA06. The expression vector pSU03-AP only possessed the minimal replication elements (rep, SSO, DSO) and exhibited more stability on structure, suggesting that the rest of the genes in pSU01 (ORF1, ORF2, mob, hsp) were unessential for the structural stability of plasmid in B. subtilis. In addition, recombinant production of the alkaline protease was achieved more efficiently with pSU03-AP whose copy number was estimated to be more than 100 per chromosome. Furthermore, pSU03-AP could also be used to transform and replicate in B. pumilus BA06 under selective pressure. In conclusion, pSU03-AP is expected to be a useful tool for gene expression in Bacillus subtilis and B. pumilus.

  19. Bacillus subtilis as potential producer for polyhydroxyalkanoates

    Directory of Open Access Journals (Sweden)

    Patel Sanjay KS

    2009-07-01

    Full Text Available Abstract Polyhydroxyalkanoates (PHAs are biodegradable polymers produced by microbes to overcome environmental stress. Commercial production of PHAs is limited by the high cost of production compared to conventional plastics. Another hindrance is the brittle nature and low strength of polyhydroxybutyrate (PHB, the most widely studied PHA. The needs are to produce PHAs, which have better elastomeric properties suitable for biomedical applications, preferably from inexpensive renewable sources to reduce cost. Certain unique properties of Bacillus subtilis such as lack of the toxic lipo-polysaccharides, expression of self-lysing genes on completion of PHA biosynthetic process – for easy and timely recovery, usage of biowastes as feed enable it to compete as potential candidate for commercial production of PHA.

  20. Morphologies and phenotypes in Bacillus subtilis biofilms.

    Science.gov (United States)

    Wang, Xiaoling; Meng, Shuo; Han, Jingshi

    2017-08-01

    In this study, we explored Bacillus subtilis biofilm growth under various conditions such as the use of substrates with different stiffnesses and nutrient levels using a well-developed optical imaging technique to spatially and temporally track biofilm growth. We also developed a quantitative method to characterize B. subtilis biofilm morphologies under various growth conditions. To determine biofilm rim irregularities, we used the dimensionless P2A ratio, defined as P 2 /4πA, where P is the perimeter and A is the area of the biofilm. To estimate biofilm thickness from transmission images, we developed a calibration procedure based on Beer- Lambert's law and cross sectioning. Furthermore, to determine the distributions of different B. subtilis cell phenotypes during biofilm growth, we used a triple-fluorescence-labeled B. subtilis strain that expressed motility, matrix production, and sporulation. Based on this work, we are able to tune biofilm growth by changing its growing environment.

  1. Germination Requirements of Bacillus macerans Spores

    Science.gov (United States)

    Sacks, L. E.; Thompson, P. A.

    1971-01-01

    2-Phenylacetamide is an effective germinant for spores of five strains of Bacillus macerans, particularly in the presence of fructose. Benzyl penicillin, the phenyl acetamide derivative of penicillin, and phenylacetic acid are also good germinants. l-Asparagine is an excellent germinant for four strains. α-Amino-butyric acid is moderately effective. Pyridoxine, pyridoxal, adenine, and 2,6-diaminopurine are potent germinants for NCA strain 7X1 only. d-Glucose is a powerful germinant for strain B-70 only. d-Fructose and d-ribose strongly potentiate germination induced by other germinants (except l-asparagine) but have only weak activity by themselves. Niacinamide and nicotinamide-adenine dinucleotide, inactive by themselves, are active in the presence of fructose or ribose. Effects of pH, ion concentration, and temperature are described. PMID:4251279

  2. Protein engineering of cyclodextrin glycosyltransferase from Bacillus circulans strain 251

    NARCIS (Netherlands)

    Penninga, Dirk

    1996-01-01

    An enormous diversity of molecular functions in living organisms is carried out by proteins. Our studies have focussed on the functional analysis of a starch-converting enzyme, cyclodextrin glycosyltransferase (CGTase) from Bacillus circulans strain 251. Zie: Summary

  3. BOOK REVIEW: BACILLUS THURINGIENSIS: A CORNERSTONE OF MODERN AGRICULTURE

    Science.gov (United States)

    Are you interested in the technical issues surrounding the use of Bacillus thuringiensis pesticidal traits as sprays and as plant incorporated protectants (transgenic crops)? Should the dimensions of human health, ecology, entomology, risk assessment, resistance management, and d...

  4. Isolation of protease producing novel Bacillus cereus and detection ...

    African Journals Online (AJOL)

    user

    2011-02-14

    1991). Industrial enzymes-developments in production and application, Biotechnol. Adv. 9: 643-658. Ferrero MA, Castro GR, Abate CM, Baigori MD, Sineriz F (1996). Thermostable alkaline proteases of Bacillus licheniformis MIR ...

  5. Determination of Levan from Bacillus licheniformis by Ultraviolet ...

    African Journals Online (AJOL)

    Keywords: Derivative spectrophotometry, Bacillus licheniformis, Levan, Total carbohydrate determination. Tropical Journal of Pharmaceutical Research is indexed by Science Citation Index (SciSearch), Scopus,. International Pharmaceutical Abstract, Chemical Abstracts, Embase, Index Copernicus, EBSCO, African.

  6. Two Genes Encoding Uracil Phosphoribosyltransferase Are Present in Bacillus subtilis

    DEFF Research Database (Denmark)

    Martinussen, Jan; Glaser, Philippe; Andersen, Paal S.

    1995-01-01

    Uracil phosphoribosyltransferase (UPRTase) catalyzes the key reaction in the salvage of uracil in many microorganisms. Surprisingly, two genes encoding UPRTase activity were cloned from Bacillus subtilis by complementation of an Escherichia coli mutant. The genes were sequenced, and the putative...

  7. Dendritic Cells Endocytose Bacillus Anthracis Spores: Implications for Anthrax Pathogenesis

    National Research Council Canada - National Science Library

    Brittingham, Katherine C; Ruthel, Gordon; Panchal, Rekha G; Fuller, Claudette L; Ribot, Wilson J

    2005-01-01

    Phagocytosis of inhaled Bacillus anthracis spores and subsequent trafficking to lymph nodes are decisive events in the progression of inhaled anthrax because they initiate germination and dissemination of spores...

  8. Analysis of Bacillus Globigii Spores Using the BioDetector

    National Research Council Canada - National Science Library

    Lee, William

    1999-01-01

    .... An automated immunoassay instrument capable of providing rapid identification of biological agents was used to analyses laboratory and field trial samples containing the field trial simulants Bacillus globigii (BG) spores...

  9. Effects of probiotic Bacillus species in aquaculture – An overview

    Directory of Open Access Journals (Sweden)

    Cristian-Teodor BURUIANĂ

    2014-12-01

    Full Text Available The ingestion of a large amount of certain types of beneficial bacteria can reduce the multiplication and development of pathogenic bacteria in the gut. A “probiotic” is a product that contains live microorganisms which positively influence the host intestinal microbiota by preventing the proliferation of pathogenic bacteria and promoting the growth and development of beneficial bacteria. Bacillus spp. are Gram-positive endospore-forming bacteria with beneficial effects in aquaculture industry. The dietary supplementation of Bacillus spp. in fish culture improved especially growth performance, immune response and the disease resistance of fish against pathogenic bacterial infections. The objective of the current paper is to review the recent published investigations reported in the scientific literature on the use of probiotic Bacillus spp. in aquaculture, focusing on their beneficial effects on the host. This review includes the main effects of Bacillus spp. administration in shrimp culture, carp culture, tilapia culture, and other fish culture.

  10. Application of the biosurfactants produced by Bacillus spp. (SH 20 ...

    African Journals Online (AJOL)

    Application of the biosurfactants produced by Bacillus spp. (SH 20 and SH 26) and P. aeruginosa SH 29 isolated from the rhizosphere soil of an Egyptian salt marsh plant for the cleaning of oil - contaminataed vessels and enhancing the biodegradat.

  11. Detoxification of cyanides in cassava flour by linamarase of Bacillus ...

    African Journals Online (AJOL)

    enoh

    2012-04-05

    Ugwuanyi et al., 2007). Effect of enzymatic treatment on cassava flour. An aliquot of 500 .... depends exclusively on endogenous enzyme level which shows considerable variation. Several microorganisms including Bacillus sp. (Amoa-.

  12. The promotive effect of N 2 fixers, Bacillus circulans and ...

    African Journals Online (AJOL)

    The promotive effect of N 2 fixers, Bacillus circulans and Saccharomyces cerevisiae on the viability of native arbuscular mycorrhizal fungi and the impact on the productivity of alfalfa ( Medicago sativa l.)

  13. Draft Genome Sequence of Bacillus amyloliquefaciens B-1895

    OpenAIRE

    Karlyshev, Andrey V.; Melnikov, Vyacheslav G.; Chistyakov, Vladimir A.

    2014-01-01

    In this report, we present a draft genome sequence of Bacillus amyloliquefaciens strain B-1895. Comparison with the genome of a reference strain demonstrated similar overall organization, as well as differences involving large gene clusters.

  14. Intramembrane protease RasP boosts protein production in Bacillus

    NARCIS (Netherlands)

    Neef, Jolanda; Bongiorni, Cristina; Goosens, Vivianne J.; Schmidt, Brian; van Dijl, Jan Maarten

    2017-01-01

    Background: The microbial cell factory Bacillus subtilis is a popular industrial platform for high- level production of secreted technical enzymes. Nonetheless, the effective secretion of particular heterologous enzymes remains challenging. Over the past decades various studies have tackled this

  15. Diversity and enzymatic characterization of Bacillus species isolated ...

    African Journals Online (AJOL)

    , phosphatases, lipases and proteases. Seventeen enzymes from these different enzyme groups were synthesized by the identified Bacillus species. The dominant and enzyme-producing species could be used for the development of a starter ...

  16. Cytotoxic effects of delfin insecticide ( Bacillus thuringiensis ) on cell ...

    African Journals Online (AJOL)

    Cytotoxic effects of delfin insecticide ( Bacillus thuringiensis ) on cell behaviour, phagocytosis, contractile vacuole activity and macronucleus in a protozoan ciliate Paramecium caudatum. ... macronucleus, fragmentation, vacuolization and complete diffusion of macronucleus were observed and were dose dependent.

  17. Proposal of nine novel species of the Bacillus cereus group.

    Science.gov (United States)

    Liu, Yang; Du, Juan; Lai, Qiliang; Zeng, Runying; Ye, Dezan; Xu, Jun; Shao, Zongze

    2017-08-01

    Nine novel Gram-stain-positive bacteria were investigated by a polyphasic taxonomic approach. Based on the analysis of 16S rRNA gene sequences, these strains belonged to the Bacillus cereus group, sharing over 97 % similarity with the known species of this group, and less than 95 % similarity with other species of the genus Bacillus. Multilocus sequence typing analysis showed that they formed nine robust and well-separated branches from the known species. The digital DNA-DNA hybridization (dDDH) and average nucleotide identity (ANI) values between the nine strains were, respectively, below the 70 and 96 % threshold values for species definition, and between each strain and the known type strains of this group were also below the two threshold values. On the basis of the phenotypic and phylogenetic data, along with low dDDH and ANI values among these strains, these bacteria are assigned to the following nine novel species of the B. cereus group: Bacillus paranthracis sp. nov., type strain Mn5T (=MCCC 1A00395T=KCTC 33714T=LMG 28873T); Bacillus pacificus sp. nov., type strain EB422T (=MCCC 1A06182T=KCTC 33858T); Bacillus tropicus sp. nov., type strain N24T (=MCCC 1A01406T=KCTC 33711T=LMG 28874T); Bacillus albus sp. nov., type strain N35-10-2T (=MCCC 1A02146T=KCTC 33710T=LMG 28875T); Bacillus mobilis sp. nov., type strain 0711P9-1T (=MCCC 1A05942T=KCTC 33717T=LMG 28877T); Bacillus luti sp. nov., type strain TD41T (=MCCC 1A00359T=KCTC 33716T=LMG 28872T); Bacillus proteolyticus sp. nov., type strain TD42T (=MCCC 1A00365T=KCTC 33715T=LMG 28870T); Bacillus nitratireducens sp. nov., type strain 4049T (=MCCC 1A00732T=KCTC 33713T=LMG 28871T); and Bacillus paramycoides sp. nov., type strain NH24A2T (=MCCC 1A04098T=KCTC 33709T=LMG 28876T).

  18. Resistance of Bacillus Endospores to Extreme Terrestrial and Extraterrestrial Environments

    Science.gov (United States)

    Nicholson, Wayne L.; Munakata, Nobuo; Horneck, Gerda; Melosh, Henry J.; Setlow, Peter

    2000-01-01

    Endospores of Bacillus spp., especially Bacillus subtilis, have served as experimental models for exploring the molecular mechanisms underlying the incredible longevity of spores and their resistance to environmental insults. In this review we summarize the molecular laboratory model of spore resistance mechanisms and attempt to use the model as a basis for exploration of the resistance of spores to environmental extremes both on Earth and during postulated interplanetary transfer through space as a result of natural impact processes. PMID:10974126

  19. Bacillus As Potential Probiotics: Status, Concerns, and Future Perspectives

    Directory of Open Access Journals (Sweden)

    Fouad M. F. Elshaghabee

    2017-08-01

    Full Text Available Spore-forming bacilli are being explored for the production and preservation of food for many centuries. The inherent ability of production of large number of secretory proteins, enzymes, antimicrobial compounds, vitamins, and carotenoids specifies the importance of bacilli in food chain. Additionally, Bacillus spp. are gaining interest in human health related functional food research coupled with their enhanced tolerance and survivability under hostile environment of gastrointestinal tract. Besides, bacilli are more stable during processing and storage of food and pharmaceutical preparations, making them more suitable candidate for health promoting formulations. Further, Bacillus strains also possess biotherapeutic potential which is connected with their ability to interact with the internal milieu of the host by producing variety of antimicrobial peptides and small extracellular effector molecules. Nonetheless, with proposed scientific evidences, commercial probiotic supplements, and functional foods comprising of Bacillus spp. had not gained much credential in general population, since the debate over probiotic vs pathogen tag of Bacillus in the research and production terrains is confusing consumers. Hence, it’s important to clearly understand the phenotypic and genotypic characteristics of selective beneficial Bacillus spp. and their substantiation with those having GRAS status, to reach a consensus over the same. This review highlights the probiotic candidature of spore forming Bacillus spp. and presents an overview of the proposed health benefits, including application in food and pharmaceutical industry. Moreover, the growing need to evaluate the safety of individual Bacillus strains as well as species on a case by case basis and necessity of more profound analysis for the selection and identification of Bacillus probiotic candidates are also taken into consideration.

  20. Potensi Bacillus Coagulans Dari Serasah Hutan Sebagai Probiotik Ayam Broiler

    OpenAIRE

    Wizna, Wizna; Abbas, H; Dharma, A; Kompiang, P

    2013-01-01

    Probiotics are living microorganisms which controls the balance of pathogenic microbes in the digestive tract of cattle through competitive exclusion mechanism which lately has been widely used as a feed aditive both ruminants and poultry . One type of microbes used in probiotics in poultry livestock is a bacterium of the genus Bacillus . Bacillus coagulans (Lactobacillus sporogenes) had the same function as Lactobacillus sp known as probiotics were able to live in the digestive tract and pro...