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Registros recuperados: 32
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A simple and efficient method for extraction of Taq DNA polymerase Electron. J. Biotechnol.
Chen,Sique; Zheng,Xiujuan; Cao,Hongrui; Jiang,Linghui; Liu,Fangqian; Sun,Xinli.
Background Thermostable DNA polymerase (Taq Pol ?) from Thermus aquaticus has been widely used in PCR, which was usually extracted with Pluthero's method. The method used ammonium sulfate to precipitate the enzyme, and it saved effort and money but not time. Moreover, we found that 30-40% activity of Taq Pol I was lost at the ammonium sulfate precipitation step, and the product contained a small amount of DNA. Results We provided a novel, simplified and low-cost method to purify the Taq Pol ? after overproduction of the enzyme in Escherichia coli, which used ethanol instead of ammonium sulfate to precipitate the enzyme. The precipitate can be directly dissolved in the storage buffer without dialysis. In addition, DNA and RNA contamination was removed with...
Tipo: Journal article Palavras-chave: Ethanol precipitation; PCR; Purification; Taq DNA polymerase.
Ano: 2015 URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582015000500005
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A single-step purification of bothropstoxin-1 BJMBR
Spencer,P.J.; Aird,S.D.; Boni-Mitake,M.; Nascimento,N.; Rogero,J.R..
Bothrops venoms are complex mixtures of components with a wide range of biological activities. Among these substances, myotoxins have been investigated by several groups. Bothropstoxin-1 (Bthtx-1) is a phospholipase A2-like basic myotoxin from Bothrops jararacussu. The purification of this component involves two chromatographic steps. Although providing a pure material, the association of these two steps is time consuming and a single-step method using high performance chromatography media would be useful. In the present study, we describe a single-step purification method for Bthtx-1. Bothrops jararacussu venom was dissolved in 1 ml buffer. After centrifugation, the supernatant was injected into a Resource-S cation exchange column connected to an FPLC...
Tipo: Info:eu-repo/semantics/other Palavras-chave: Myotoxins; Purification; Chromatography; Bothrops jararacussu; Bothropstoxin-1.
Ano: 1998 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X1998000900004
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Bactéries pathogènes indigènes des eaux estuariennes: cas des vibrios. Revue bibliographique ArchiMer
Fouche, Dominique.
De nombreuses bactéries, peuplant habituellement les zones estuariennes, se révèlent être de plus en plus fréquemment des agents de maladies dont les vecteurs principaux sont les coquillages et leur consommation par l'homme. Le présent travail synthétise l'état des connaissances sur les vibrios, en particulier Vibrio cholerae, Vibrio parahaemolyticus et Vibrio vulnificus, bactéries pour lesquelles il existe une  abondante littérature. L'écologie, le pouvoir pathogène et les milieux de culture les plus couramment utilisés en vue de leur détection sont présentés. Enfin, leur interaction avec les mollusques bivalves, leur survie dans ceux-ci et le problème de l'épuration conséquente de ces derniers sont abordés.
Tipo: Text Palavras-chave: Vibrio cholerae; Vibrio parahaemolyticus; Vibrio vulnificus; Bactéries indigènes; Estuaires; Bivalves; Purification.
Ano: 1993 URL: http://archimer.ifremer.fr/doc/00408/51929/52554.pdf
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Characterization of bacteriocin-like substances synthesized by Parabacteroides distasonis strains isolated from broiles feces. Repositório Alice
OLIVEIRA, A. G. G.; OLIVEIRA, J. S.; OLIVEIRA, P. L.; CARVALHO, M. A. R.; NICOLI, J. R.; BEMQUERER, M. P.; MAGALHÃES, P. P.; FARIAS, L. M..
2015
Tipo: Resumo em anais de congresso (ALICE) Palavras-chave: Antagonistic substance; Bacteriocin; Parabacteroides distasonis; Broiler chicken; Purification.
Ano: 2015 URL: http://www.alice.cnptia.embrapa.br/handle/doc/1027493
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Chitinase from Enterobacter sp. NRG4: Its purification, characterization and reaction pattern Electron. J. Biotechnol.
Dahiya,Neetu; Tewari,Rupinder; Tiwari,Ram P; Singh Hoondal,Gurinder.
Enterobacter sp. NRG4 was shown to excrete chitinase into the culture supernatant when cultivated in medium containing chitin. A 60 kDa extracellular chitinase was purified to homogeneity and characterized. The enzyme hydrolyzed swollen chitin, colloidal chitin, regenerated chitin and glycol chitin but did not hydrolyze chitosan. The chitinase exhibited Km and Vmax values of 1.43 mg ml-1 and 83.33 µM µg-1 h-1 for swollen chitin, 1.41 mg ml-1 and 74.07 µM µg-1 h-1 for colloidal chitin, 1.8 mg ml-1 and 40 µM µg-1 h-1 for regenerated chitin and 2.0 mg ml-1 and 33.33 µM µg-1 h-1 for glycol chitin, respectively. The optimal temperature and pH for activity were 45ºC and pH 5.5, respectively. Mg2+, K+ and Ca2+ stimulated chitinase activity by 13, 16 and 18%,...
Tipo: Journal article Palavras-chave: Chemical modification; Chitinase; Enterobacter sp. NRG4; Purification; Substrate binding.
Ano: 2005 URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582005000200003
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Comparison of commercially-available RNA extraction methods for effective bacterial RNA isolation from milk spiked samples Electron. J. Biotechnol.
García-Nogales,Paula; Serrano,Alicia; Secchi,Sonia; Gutiérrez,Serafín; Arís,Anna.
Nucleic-acid based methods for bacterial identification are extremely useful in diagnostic applications due to their specificity and sensitivity. However, they require an optimal purification of the target molecules. As part of the development of a new diagnostic method for the detection of bacterial RNA in cow milk, we have compared four commercially available RNA extraction kits for the isolation of bacterial RNA from spiked UHT milk samples. The kits were compared in terms of extraction efficiency and RNA purity using two bacterial species, the Gram negative Escherichia coli and the Gram positive Staphylococcus aureus. Two kits are based in silica-matrix extraction, and the other two in the guanidinium thiocyanate-phenol-chloroform extraction. In our...
Tipo: Journal article Palavras-chave: Bacteria; Milk; Purification; RNA.
Ano: 2010 URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582010000500019
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Depuration centre management. Course notes ArchiMer
O'Carroll, Terence; Boulter, Mark; Seaver, Mary; Masson, Daniel; O'Driscoll, Michael; Roden, Cillian.
The purpose of this course is to train and familiarise people, who are going to build, have just built or are about to modernise a purification plant, in the latest technology available to the industry and also to train people so that they can effectively run a depuration centre. ln general the course aims to : - Give participants practical experience of different systems prior to committing themselves to a particular system. - Enable participants to compare systems. - Provide a minimum training qualification to run an establishment which is essentially a facility to guarantee health and safety of products to the consumer. - Provide an overview from bacteria testing to plant lay out etc. The structure of the course is in modules, morning and...
Tipo: Text Palavras-chave: Shellfish depuration; Purification; Bacteria; Viruses.
Ano: 1995 URL: http://archimer.ifremer.fr/doc/00140/25082/23192.pdf
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Essais de décontamination bactériologique des huîtres Crassostrea gigas en bassins par renouvellement et circulation d'eau ArchiMer
Faury, Nicole; Ratiskol, Jacqueline.
La décontamination bactériologique des huitres Crassostrea gigas a été étudiée expérimentalement dans des bassins dégorgeoirs de 13 m3 de conception proche de la configuration normale d'une station de purification française. Les huîtres contaminées artificiellement ont été placées dans des conditions dites "naturelles" d'épuration. Il n'a pas été utilisé de procédé physique ou chimique de purification de l'eau. Elles ont été soumises sur trois périodes de l'année à trois conditions différentes de renouvellement d'eau : quotidien, biquotidien ou apport continu, le témoin étant caractérisé par l'absence de tout renouvellement d'eau. Les résultats permettent de dégager un ensemble de remarques sur l'état physiologique, sur l'activité métabolique des...
Tipo: Text Palavras-chave: Décontamination; Coliformes fécaux; Streptocoques fécaux; Bassins; Purification.
Ano: 1993 URL: http://archimer.ifremer.fr/doc/00050/16156/13639.pdf
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Etude de l'herpès virus de l'huître. Fonds d'Aide à la Recherche et à l'Innovation. Décision d'Aide à la Recherche n° 95-07-01 (Rapport intermédiaire) ArchiMer
Renault, Tristan.
L'Unité de Recherche en Pathologie et Immunologie Générales (Station IFREMER de La Tremblade) se propose dans le présent projet d'étudier le virus de type herpès observée chez les huîtres (pouvoir pathogène, conditions d'expression de l'infection, transmission, etc ... ) et de développer des réactifs permettant de réaliser un diagnostic rapide, sensible et fiable de l'infection qu'il occassionne. Les diagnostics proposés seront basés sur la détection directe du virus, soit au moyen de réactifs immunologiques spécifiques des antigènes viraux, soit par des techniques de biologie moléculaire permettant de détecter l'ADN viral (hybridation de sondes nucléiques et réaction PCR : Polymerase Chain Reaction). Ce projet, proposé sur une durée de trois années...
Tipo: Text Palavras-chave: Virus; Herpès; Pathogène; Purification; Herpesviridae; Crassostrea gigas.
Ano: 1996 URL: http://archimer.ifremer.fr/doc/00043/15447/12818.pdf
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Fermentation and recovery of L-glutamic acid from cassava starch hydrolysate by ion-exchange resin column Rev. Microbiol.
Nampoothiri,K. Madhavan; Pandey,Ashok.
Investigations were carried out with the aim of producing L-glutamic acid from Brevibacterium sp. by utilizing a locally available starchy substrate, cassava (Manihot esculenta Crantz). Initial studies were carried out in shake flasks, which showed that even though the yield was high with 85-90 DE (Dextrose Equivalent value), the maximum conversion yield (~34%) was obtained by using only partially digested starch hydrolysate, i.e. 45-50 DE. Fermentations were carried out in batch mode in a 5 L fermenter, using suitably diluted cassava starch hydrolysate, using a 85-90 DE value hydrolysate. Media supplemented with nutrients resulted in an accumulation of 21 g/L glutamic acid with a fairly high (66.3%) conversation yield of glucose to glutamic acid (based on...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Brevibacterium sp.; L-glutamic acid; Cassava hydrolysate; Batch and fed-batch process; Ion-exchange resin; Purification.
Ano: 1999 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0001-37141999000300013
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Gene expression and characterization of 2-keto-3-deoxy-gluconate kinase, a key enzyme in the modified Entner-Doudoroff pathway of Serratia marcescens KCTC 2172 Electron. J. Biotechnol.
Lee,Yong-Seok; Park,In-Hye; Yoo,Ju-Soon; Kim,Hae-Sun; Chung,Soo-Yeol; Chandra,Muni Ramanna GariSubhosh; Choi,Yong-Lark.
We cloned 2-keto-3-deoxy-gluconate kinase (KDGK), which catalyzes the phosphorylation of 2-keto-3-deoxygluconate (KDG) to 2-keto-3-deoxy-6-phophogluconate (KDPG) from Serratia marcescens KCTC 2172. The nucleotide sequence revealed a single open reading frame containing 1,208 bp and encoding for 309 amino acids, with a molecular weight of 33,993 Da. The enzyme was purified via GST affinity chromatography. The putative KdgT binding site was detected upstream of the initial codon. The KDG kinase utilized 2-ketogluconate (KG) and KDG as substrates. The optimal temperature and pH for KDGK activity were 50ºC and 8.0, respectively.
Tipo: Journal article Palavras-chave: 2-keto-3-deoxygluconate kinase; Carbohydrate kinase; Purification; Serratia marcescens KCTC 2172.
Ano: 2009 URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582009000300005
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Les systèmes de solvants en chromatographie de partage centrifuge ArchiMer
Camacho-frias, E; Foucault, A.
Centrifugal partition chromatography (CPC) or countercurrent chromatography (CCC) is a kind of liquid-liquid chromatography without solid phase. Two immiscible liquids in equilibrium share the total volume of a ''column'' designed to keep one phase pseudo-stationary. A tremendous variety of biphasic systems are available, which can lead to some difficulty when a selection must be made. Ternary diagrams, describing mixtures of three solvents, and systems with four solvents or more, used by research scientists familiar with this chromatographic technique, are useful guides for a rational choosing of a biphasic system in CCC & CPC.
Tipo: Text Palavras-chave: Countercurrent chromatography; Centrifugal partition chromatography; Ternary diagram; Separation; Purification.
Ano: 1996 URL: http://archimer.ifremer.fr/doc/00282/39325/57898.pdf
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Ovatoxines: Purification à partir d’Ostreopsis cf. ovata en culture et niveaux d’accumulation dans les produits de la mer ArchiMer
Brissard, Charline.
Dinoflagellates of genus Ostreopsis are known in tropical areas to provoke food intoxication, after palytoxin accumulation in seafood. Probably in link with climatic changes, Ostreopsis sp. is increasingly present in French Mediterranean since 2006. In the context of the PhD, the studies in natural environment, in summer 2011 and 2012, showed that: i) Ostreopsis cf. ovata strain presented a complex toxinic profile, including palytoxin and especially ovatoxins (OVTX-a, -b, c, -d, -e). A new analog (OVTX-h) was detected; ii) cell and toxinic concentrations decreased with depth; iii) different seafood accumulated toxins, which could represent real risk for bathers and consumers. Because there are no standards for OVTXs, one of the PhD goals was to develop a...
Tipo: Text Palavras-chave: Ostreopsis ovata; Ovatoxines; Purification; Contamination des produits de la mer; Chromatographie liquide/spectrométrie de masse.; Ostreopsis ovata; Ovatoxins; Purification; Seafood contamination; Liquid chromatography / mass spectrometry..
Ano: 2014 URL: http://archimer.ifremer.fr/doc/00255/36613/35179.pdf
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Pollen as a means of international transfer of germplasm Open Agri
Mallikarjuna, N.,( Nalini Mallikarjuna ).
Palavras-chave: Chickpeas; Sucrose; Self pollination; Germplasm; Crossing over; Grain; Developmental stages; Purification; Organic compounds; Legumes.
Ano: 2007 URL: http://agropedia.iitk.ac.in/openaccess/?q=node/3290
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Pre-purification by membrane filtration of paralytic shellfish toxins from Alexandrium minutum dinoflagellate ArchiMer
Balti, Rafik; Brodu, Nicolas; Zhang, Jiaxuan; Amzil, Zouher; Drouin, Delphine; Sechet, Veronique; Masse, Anthony.
The Paralytic shellfish neurotoxins (PST) are of increasing interest for biomedical applications. The chemical synthesis is often complex and expensive that’s why the purification by membrane filtration of PST from Alexandrium minutum dinoflagellate was investigated. Disrupted micro-alga cells by ultrasonic treatment were diafiltred to let pass toxins through an ultrafiltration membrane. Then, the mean permeate was concentrated and diafiltrated by nanofiltration. Mean permeate fluxes equal to 187, 135 and 135 L.h–1.m-2 were obtained during the first diafiltration, the concentration step and the final diafiltration respectively. Up to 57 % (mol/mol) and 78 % (mol/mol) of organic matters and salts were removed respectively. Divalent ions were sparsely...
Tipo: Text Palavras-chave: Paralytic shellfish poisoning; Nanofiltration; Membrane; Alexandrium minutum; Purification.
Ano: 2019 URL: https://archimer.ifremer.fr/doc/00451/56286/57865.pdf
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Production of recombinant enzymes of wide use for research Electron. J. Biotechnol.
Manzur,María J.; Muñoz,Rosana V.; Lucero,Adrián A.; Juri Ayub,Maximiliano; Alvarez,Sergio E.; Ciuffo,Gladys M..
For biotechnological purposes, protein expression refers to the directed synthesis of large amounts of desired proteins. The aim of the present work was to produce reverse transcriptase Moloney murine Leukaemia Virus retro-transcriptase and Taq DNA polymerase, as bioactive products. In the present paper, we report the preparation of recombinant enzymes, expressed in E. coli strains. The enzymes produced exhibited quite good activity, compared with commercial enzymes, allowing us to replace the last ones for several lab applications. We are reporting changes and modifications to standard protocols described. The standard protocols were modified, i.e. for the purification step of Taq, a temperature dependent procedure was designed. The enzymes produced were...
Tipo: Journal article Palavras-chave: Bioactivity; Protein expression; Purification; Recombinant enzymes.
Ano: 2006 URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582006000300023
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Production, purification and characterization of recombinant human antithrombin III by Saccharomyces cerevisiae Electron. J. Biotechnol.
Mallu,Maheswara Reddy; Vemula,Sandeep; Ronda,Srinivasa Reddy.
Background: Antithrombin III (ATIII) is a protein that inhibits abnormal blood clots (or coagulation) by breaking down thrombin and factor Xa. ATIII helps to keep a healthy balance between hemorrhage and coagulation. The present work demonstrated the production, purification and characterization of recombinant human antithrombin (rhAT) from yeast Saccharomyces cerevisiae BY4741 was demonstrated. After expression of rhAT by S. cerevisiae, the biomass and rhAT concentration were analyzed through fed-batch fermentation process. Results: In fed-batch fermentation, the biomass (maximum cell dry weight of 11.2 g/L) and rhAT concentration (312 mg/L) of the expressed rhAT were achieved at 84 h of cultivation time. The maximum cell lysis efficiency (99.89%) was...
Tipo: Journal article Palavras-chave: Biological activity; Cell lysis; Cross flow filtration; Fed-batch fermentation; Purification; Secondary structure.
Ano: 2016 URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582016000400012
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Programme Pathologie - Rapport scientifique Année 1994 - Contrat de Plan Etat Région Poitou-Charentes 1994-1998 - Convention 94 RPC R 62 ArchiMer
Renault, Tristan; Cochennec, Nathalie; Le Deuff, Rose-marie; Chollet, Bruno; Maffart, Patricia; Haffner, Philippe; Berthe, Franck; Thuillier, Benoit.
Le but de ce programme pathologie est de recherher et d'étudier les agents pathogènes connus et inconnus chez les mollusques bivalves marins, plus particulièrement chez l'huître creuse, Crassastrea gigas, afin d'identifier d'éventuels agents causant des mortalité, de proposer des mesures prophylactiques et de définir des conditions zootechniques permettant aux professionnels de limiter l'impact des maladies sur leur cheptel. La présence de maladies, chez les mollusques bivalves marins dont l'impact socioéconomique peut être catastrophique pour les productions conchylicoles, et l'accroissement des échanges de produits d'aquaculture entre les pays de la CEE et avec les pays tiers nécessitent le renforcement des recherches dans les domaines de la pathologie....
Tipo: Text Palavras-chave: Pathologie; Agent pathogène; Infection; Virus; Herpèsvirus; Purification; Huître creuse; Crassostrea gigas.
Ano: 1994 URL: http://archimer.ifremer.fr/doc/00044/15502/12874.pdf
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Purification and characterization of infectious myonecrosis virus of penaeid shrimp ArchiMer
Poulos, Bonnie; Tang, K; Pantoja, C; Bonami, Jean-robert; Lightner, D.
The causative agent of myonecrosis affecting cultured Penaeus vannamei in Brazil was demonstrated to be a virus after purification of the agent from infected shrimp tissues. Purified viral particles were injected into specific pathogen-free P. vannamei, resulting in a disease that displayed the same characteristics as those found in the original shrimp used for purification. The virus was named infectious myonecrosis virus (IMNV). The viral particles were icosahedral in shape and 40 nm in diameter, with a buoyant density of 1(.)366 g ml(-1) in caesium chloride. The genome consisted of a single, double-stranded (dsRNA) molecule of 7560 bp. Sequencing of the viral genome revealed two non-overlapping open reading frames (ORFs). The 5' ORF (ORF 1, nt 136-4953)...
Tipo: Text Palavras-chave: Purification; Myonecrosis; Virus; Shrimp; Penaeid.
Ano: 2006 URL: http://archimer.ifremer.fr/doc/2006/publication-1504.pdf
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Purification and characterization of ß-glucosidase from Melanocarpus sp. MTCC 3922 Electron. J. Biotechnol.
Kaur,Jatinder; Chadha,Bhupinder S; Kumar,Badhan A; Kaur,Ghatora, S; Saini,Harvinder S.
This study reports the purification and characterization of ß-glucosidase from a newly isolated thermophilic fungus, Melanocarpus sp. Microbial Type Culture Collection (MTCC) 3922. The molecular weight of ß-glucosidase was determined to be ~ 92 and 102 kDa with SDS PAGE and gel filtration, respectively, and pI of ~ 4.1. It was optimally active at 60ºC and pH 6.0, though was stable at 50ºC and pH 5.0 - 6.0. The presence of DTT, mercaptoethanol and metal ions such as Na+, K+, Ca2+, Mg2+and Zn2+ positively influenced the activity of ß-glucosidase but the activity was inhibited in the presence of CuSO4. ß-Glucosidase recognized pNP- ß-glucopyranoside (pNPG) as the preferred substrate, and showed very low affinity for pNP- ß-D-cellobioside. Km and Vmax for the...
Tipo: Journal article Palavras-chave: SS-glucosidase; Melanocarpus sp; Purification; Substrate specificity; Transglycosylation.
Ano: 2007 URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582007000200010
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