Atenção: Todas as denúncias são sigilosas e sua identidade será preservada.
Os campos nome e e-mail são de preenchimento opcional
Metadados | Descrição | Idioma |
---|---|---|
Autor(es): dc.contributor | Universidade Estadual Paulista (Unesp) | - |
Autor(es): dc.contributor | Universidade de São Paulo (USP) | - |
Autor(es): dc.creator | de Moura, Werner Alfinito Feio [UNESP] | - |
Autor(es): dc.creator | Schultz, Leonardo [UNESP] | - |
Autor(es): dc.creator | Breyer, Carlos Alexandre [UNESP] | - |
Autor(es): dc.creator | de Oliveira, Ana Laura Pires [UNESP] | - |
Autor(es): dc.creator | Tairum, Carlos Abrunhosa [UNESP] | - |
Autor(es): dc.creator | Fernandes, Gabriella Costa [UNESP] | - |
Autor(es): dc.creator | Toyama, Marcos Hikari [UNESP] | - |
Autor(es): dc.creator | Pessoa-Jr, Adalberto | - |
Autor(es): dc.creator | Monteiro, Gisele | - |
Autor(es): dc.creator | de Oliveira, Marcos Antonio [UNESP] | - |
Data de aceite: dc.date.accessioned | 2022-02-22T00:34:57Z | - |
Data de disponibilização: dc.date.available | 2022-02-22T00:34:57Z | - |
Data de envio: dc.date.issued | 2020-12-11 | - |
Data de envio: dc.date.issued | 2020-12-11 | - |
Data de envio: dc.date.issued | 2019-12-31 | - |
Fonte completa do material: dc.identifier | http://dx.doi.org/10.1007/s10529-020-02955-5 | - |
Fonte completa do material: dc.identifier | http://hdl.handle.net/11449/201941 | - |
Fonte: dc.identifier.uri | http://educapes.capes.gov.br/handle/11449/201941 | - |
Descrição: dc.description | Acute lymphoblastic leukaemia (ALL) affects lymphoblastic cells and is the most common neoplasm during childhood. Among the pharmaceuticals used in the treatment protocols for ALL, Asparaginase (ASNase) from Escherichia coli (EcAII) is an essential biodrug. Meanwhile, the use of EcAII in neoplastic treatments causes several side effects, such as immunological reactions, hepatotoxicity, neurotoxicity, depression, and coagulation abnormalities. Commercial EcAII is expressed as a recombinant protein, similar to novel enzymes from different organisms; in fact, EcAII is a tetrameric enzyme with high molecular weight (140 kDa), and its overexpression in recombinant systems often results in bacterial cell death or the production of aggregated or inactive EcAII protein, which is related to the formation of inclusion bodies. On the other hand, several commercial expression strains have been developed to overcome these expression issues, but no studies on a systematic evaluation of the E. coli strains aiming to express recombinant asparaginases have been performed to date. In this study, we evaluated eleven expression strains at a low temperature (16 °C) with different characteristics to determine which is the most appropriate for asparaginase expression; recombinant wild-type EcAII (rEcAII) was used as a prototype enzyme and the secondary structure content, oligomeric state, aggregation and specific activity of the enzymes were assessed. Structural analysis suggested that a correctly folded tetrameric rEcAII was obtained using ArcticExpress (DE3), a strain that co-express chaperonins, while all other strains produced poorly folded proteins. Additionally, the enzymatic assays showed high specific activity of proteins expressed by ArcticExpress (DE3) when compared to the other strains used in this work. | - |
Descrição: dc.description | Institute of Biosciences São Paulo State University (UNESP), Coastal Campus | - |
Descrição: dc.description | Biochemical-Pharmaceutical Technology Department Faculty of Pharmaceutical Sciences University of São Paulo | - |
Descrição: dc.description | Institute of Biosciences São Paulo State University (UNESP), Coastal Campus | - |
Idioma: dc.language | en | - |
Relação: dc.relation | Biotechnology Letters | - |
???dc.source???: dc.source | Scopus | - |
Palavras-chave: dc.subject | Acute lymphoblastic leukemia (ALL) | - |
Palavras-chave: dc.subject | Escherichia coli ASNaseII (EcAII) | - |
Palavras-chave: dc.subject | Functional and structural evaluation | - |
Palavras-chave: dc.subject | Low temperature expression | - |
Palavras-chave: dc.subject | Recombinant enzyme | - |
Título: dc.title | Functional and structural evaluation of the antileukaemic enzyme l-asparaginase II expressed at low temperature by different Escherichia coli strains | - |
Tipo de arquivo: dc.type | livro digital | - |
Aparece nas coleções: | Repositório Institucional - Unesp |
O Portal eduCAPES é oferecido ao usuário, condicionado à aceitação dos termos, condições e avisos contidos aqui e sem modificações. A CAPES poderá modificar o conteúdo ou formato deste site ou acabar com a sua operação ou suas ferramentas a seu critério único e sem aviso prévio. Ao acessar este portal, você, usuário pessoa física ou jurídica, se declara compreender e aceitar as condições aqui estabelecidas, da seguinte forma: