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	Provedor de dados BJMBR (4) BJM (1) Rev. Microbiol. (1) BJPS (1) Autor Abreu,P.A.E. (1) Amaro,Beatriz Panichi (1) Andrade,A.S.R. (1) Avilán,L. (1) Bastidas,M. (1) Carvalho,H. (1) Cruz,J. (1) Franco,Telma Teixeira (1) Ho,P.L. (1) Jürgensen,C. (1) Meneghini,R. (1) Meneguetti,Giovanna Pastore (1) Nascimento,A.L.T.O. (1) Pessoa-Junior,Adalberto (1) Pontes,Marcela Zanella Ribeiro (1) Puig,J. (1) Ramos,C.R.R. (1) Rangel-Yagui,Carlota Oliveira (1) Santos,João Henrique Picado Madalena (1) Silva,Daniel Pereira da (1) Mais... Palavra-chave Protein purification (7) Aqueous two-phase systems (2) Affinity (1) B-galactosidase (1) Biobetters (1) Biological drugs (1) Chaperonins (1) Downstream-processing (1) Escherichia coli (1) Expression vector (1) Gel filtration (1) Glucagon (1) Glucose 6-phosphate dehydrogenase (1) GroEL (1) GroES (1) Hexokinase (1) Immobilized metal affinity chromatography (1) Insulin (1) Ion-exchange chromatography (1) Iron metabolism (1) Mais... Tipo do documento Info:eu-repo/semantics/article (7) Ano 2018 (1) 2008 (1) 2004 (1) 2002 (1) 1999 (1) 1997 (2) Mais... País Brazil (7) Idioma Inglês (7)		Ordenar por:  Relevância Autor Título Ano Registros recuperados: 7 Primeira ... 1 ... Última A high-copy T7 Escherichia coli expression vector for the production of recombinant proteins with a minimal N-terminal His-tagged fusion peptide BJMBR Ramos,C.R.R.; Abreu,P.A.E.; Nascimento,A.L.T.O.; Ho,P.L.. We report here the construction of a vector derived from pET3-His and pRSET plasmids for the expression and purification of recombinant proteins in Escherichia coli based on T7 phage RNA polymerase. The resulting pAE plasmid combined the advantages of both vectors: small size (pRSET), expression of a short 6XHis tag at N-terminus (pET3-His) and a high copy number of plasmid (pRSET). The small size of the vector (2.8 kb) and the high copy number/cell (200-250 copies) facilitate the subcloning and sequencing procedures when compared to the pET system (pET3-His, 4.6 kb and 40-50 copies) and also result in high level expression of recombinant proteins (20 mg purified protein/liter of culture). In addition, the vector pAE enables the expression of a fusion... Tipo: Info:eu-repo/semantics/article Palavras-chave: Escherichia coli; Expression vector; Immobilized metal affinity chromatography; Protein purification. Ano: 2004 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2004000800001 Purification of bovine pancreatic glucagon as a by-product of insulin production BJMBR Andrade,A.S.R.; Vilela,L.; Tunes,H.. A process for purifying bovine pancreatic glucagon as a by-product of insulin production is described. The glucagon-containing supernatant from the alkaline crystallization of insulin was precipitated using ammonium sulfate and isoelectric precipitation. The isoelectric precipitate containing glucagon was then purified by ion-exchange chromatography on Q-Sepharose FF, gel filtration on Sephadex G-25 and ion-exchange chromatography on S-Sepharose FF. A pilot scale test was performed with a recovery of 87.6% and a purification factor of 8.78 for the first chromatographic step, a recovery of 75.1% and a purification factor of 3.90 for the second, and a recovery of 76.2% and a purification factor of 2.36 for the last one. The overall yield was 50%, a... Tipo: Info:eu-repo/semantics/article Palavras-chave: Glucagon; Protein purification; Gel filtration; Ion-exchange chromatography; Insulin. Ano: 1997 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X1997001200006 Cloning, expression and purification of recombinant streptokinase: partial characterization of the protein expressed in Escherichia coli BJMBR Avilán,L.; Yarzábal,A.; Jürgensen,C.; Bastidas,M.; Cruz,J.; Puig,J.. We cloned the streptokinase (STK) gene of Streptococcus equisimilis in an expression vector of Escherichia coli to overexpress the profibrinolytic protein under the control of a tac promoter. Almost all the recombinant STK was exported to the periplasmic space and recovered after gentle lysozyme digestion of induced cells. The periplasmic fraction was chromatographed on DEAE Sepharose followed by chromatography on phenyl-agarose. Active proteins eluted between 4.5 and 0% ammonium sulfate, when a linear gradient was applied. Three major STK derivatives of 47.5 kDa, 45 kDa and 32 kDa were detected by Western blot analysis with a polyclonal antibody. The 32-kDa protein formed a complex with human plasminogen but did not exhibit Glu-plasminogen activator... Tipo: Info:eu-repo/semantics/article Palavras-chave: Recombinant streptokinase; Plasminogen activators; Protein purification. Ano: 1997 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X1997001200007 Increased expression and purification of soluble iron-regulatory protein 1 from Escherichia coli co-expressing chaperonins GroES and GroEL BJMBR Carvalho,H.; Meneghini,R.. Iron is an essential metal for all living organisms. However, iron homeostasis needs to be tightly controlled since iron can mediate the production of reactive oxygen species, which can damage cell components and compromise the integrity and/or cause DNA mutations, ultimately leading to cancer. In eukaryotes, iron-regulatory protein 1 (IRP1) plays a central role in the control of intracellular iron homeostasis. This occurs by interaction of IRP1 with iron-responsive element regions at 5' of ferritin mRNA and 3' of transferrin mRNA which, respectively, represses translation and increases mRNA stability. We have expressed IRP1 using the plasmid pT7-His-hIRP1, which codifies for human IRP1 attached to an NH2-terminal 6-His tag. IRP1 was expressed in... Tipo: Info:eu-repo/semantics/article Palavras-chave: Iron metabolism; Iron-regulatory protein 1; Chaperonins; GroES; GroEL; Protein purification. Ano: 2008 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2008000400003 Infleunce of pH on the partition of glucose-6-phosphate dehydrogenase and hexokinase in aqueous two-phase system BJM Silva,Daniel Pereira da; Pontes,Marcela Zanella Ribeiro; Souza,Maria Aparecida de; Vitolo,Michele; Silva,João Batista de Almeida e; Pessoa-Junior,Adalberto. Glucose-6-phosphate dehydrogenase (G6PDH) and hexokinase (HK) are important enzymes used in biochemical and medical studies and in several analytical methods. Aqueous two-phase system (ATPS) formed by a polymer solution and an electrolyte solution provides a method for the separation and purification of enzymes with several advantages, including biocompatibility and easy scale up of the process. In this work, the effects of different pH values on the storage stability and partitioning behavior (K, partition coefficient) of the enzymes G6PDH and HK from baker's yeast extract were investigated in ATPS. The results, obtained from the 17.5% PEG 400 : 15.0% phosphate system, showed that when the pH was increased from 5.0 to 8.8, the K HK increased 26-fold and... Tipo: Info:eu-repo/semantics/article Palavras-chave: Aqueous two-phase systems; Partition coefficient; Protein purification; Hexokinase; Glucose 6-phosphate dehydrogenase. Ano: 2002 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822002000300002 Protein PEGylation for the design of biobetters: from reaction to purification processes BJPS Santos,João Henrique Picado Madalena; Torres-Obreque,Karin Mariana; Meneguetti,Giovanna Pastore; Amaro,Beatriz Panichi; Rangel-Yagui,Carlota Oliveira. The covalent attachment of polyethylene glycol (PEG) to therapeutical proteins is an important route to develop biobetters for biomedical, biotech and pharmaceutical industries. PEG conjugation can shield antigenic epitopes of the protein, reduce degradation by proteolytic enzymes, enhance long-term stability and maintain or even improve pharmacokinetic and pharmacodynamics characteristics of the protein drug. Nonetheless, correct information in terms of the PEGylation process from reaction to downstream processing is of paramount importance for the industrial application and processing scale-up. In this review we present and discuss the main steps in protein PEGylation, namely: PEGylation reaction, separation of the products and final characterization of... Tipo: Info:eu-repo/semantics/article Palavras-chave: PEGylation; Biobetters; Biological drugs; Polyethylene glycol; Protein purification; Site-specific PEGylation. Ano: 2018 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1984-82502018000700408 Purification of microbial b-galactosidase from Kluyveromyces fragilis by bioaffinity partitioning Rev. Microbiol. Silva,Maria Estela da; Franco,Telma Teixeira. This work investigated the partitioning of b-galactosidase from Kluyveromyces fragilis in aqueous two-phase systems (ATPS) by bioaffinity. PEG 4000 was chemically activated with thresyl chloride, and the biospecific ligand p-aminophenyl 1-thio-b-D-galactopyranoside (APGP) was attached to the activated PEG 4000. A new two-step method for extraction and purification of the enzyme b-galactosidase from Kluyveromyces fragilis was developed. In the first step, a system composed of 6% PEG 4000-APGP and 8% dextran 505 was used, where b-galactosidase was strongly partitioned to the top phase (K = 2,330). In the second step, a system formed of 13% PEG-APGP and 9% phosphate salt was used to revert the value of the partition coefficient of b-galactosidase (K = 2 x... Tipo: Info:eu-repo/semantics/article Palavras-chave: B-galactosidase; Aqueous two-phase systems; Protein purification; Downstream-processing; Affinity. Ano: 1999 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0001-37141999000400006 Registros recuperados: 7 Primeira ... 1 ... Última

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