| Texto completo | |
| Autor(es): |
de Moura, Werner Alfinito Feio
;
Schultz, Leonardo
;
Breyer, Carlos Alexandre
;
de Oliveira, Ana Laura Pires
;
Tairum, Carlos Abrunhosa
;
Fernandes, Gabriella Costa
;
Toyama, Marcos Hikari
;
Pessoa-Jr, Adalberto
;
Monteiro, Gisele
;
de Oliveira, Marcos Antonio
Número total de Autores: 10
|
| Tipo de documento: | Artigo Científico |
| Fonte: | Biotechnology Letters; v. 42, n. 11, p. 12-pg., 2020-07-07. |
| Resumo | |
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) fromEscherichia 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 theE. colistrains aiming to express recombinant asparaginases have been performed to date. In this study, we evaluated eleven expression strains at a low temperature (16 degrees 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. (AU) | |
| Processo FAPESP: | 13/08617-7 - Produção de L-asparaginase extracelular: da bioprospecção à engenharia de um biofármaco antileucêmico |
| Beneficiário: | Adalberto Pessoa Junior |
| Modalidade de apoio: | Auxílio à Pesquisa - Temático |
| Processo FAPESP: | 11/13500-6 - Investigação dos determinantes moleculares envolvidos na interação com os substratos de Tsa1 e Tsa2 de Saccharomyces cerevisiae |
| Beneficiário: | Carlos Alexandre Breyer |
| Modalidade de apoio: | Bolsas no Brasil - Doutorado |
| Processo FAPESP: | 16/19245-1 - Avaliação de cepas bacterianas na superexpressão da enzima antileucêmica L-Asparaginase de Escherichia coli |
| Beneficiário: | Werner Alfinito Feio de Moura |
| Modalidade de apoio: | Bolsas no Brasil - Iniciação Científica |
| Processo FAPESP: | 17/25272-4 - Asparaginase1 de Saccharomyces cerevisiae: Investigação dos efeitos funcionais e estruturais da cauda N-terminal |
| Beneficiário: | Gabriella Costa Fernandes |
| Modalidade de apoio: | Bolsas no Brasil - Iniciação Científica |
| Processo FAPESP: | 19/04054-4 - Busca de inibidores do sistema peroxirredoxina 2-Cys típicas de eucariotos: avaliação funcional e estrutural |
| Beneficiário: | Ana Laura Pires de Oliveira |
| Modalidade de apoio: | Bolsas no Brasil - Mestrado |
| Processo FAPESP: | 17/19942-7 - Busca de inibidores do sistema peroxirredoxina de patógenos e humanos |
| Beneficiário: | Marcos Antonio de Oliveira |
| Modalidade de apoio: | Auxílio à Pesquisa - Regular |
| Processo FAPESP: | 14/22039-9 - Caracterização funcional, estrutural e modificação racional da ASPaseM: um novo fármaco para o tratamento da anemia linfóide aguda? |
| Beneficiário: | Leonardo Schultz da Silva |
| Modalidade de apoio: | Bolsas no Brasil - Doutorado Direto |
| Processo FAPESP: | 17/20291-0 - Caracterização da atividade pró-inflamatória de uma nova serino protease (cdtsp-2) purificada do veneno total de Crotalus durissus terrificus |
| Beneficiário: | Marcos Hikari Toyama |
| Modalidade de apoio: | Auxílio à Pesquisa - Regular |