| Author | Girão, Luciana Facchinetti de Castro | |
| Author | Rocha, Surza Lucia Gonçalves da | |
| Author | Sobral, Ricardo Sposina | |
| Author | Bom, Ana Paula Dinis Ano | |
| Author | Sampaio, André Luiz Franco | |
| Author | Silva, José Godinho da | |
| Author | Ferrara, Maria Antonieta | |
| Author | Bon, Elba Pinto da Silva | |
| Author | Perales, Jonas | |
| Access date | 2016-07-14T13:53:36Z | |
| Available date | 2016-07-14T13:53:36Z | |
| Document date | 2016 | |
| Citation | GIRÃO, Luciana Facchinetti de Castro; et al. Saccharomyces cerevisiae asparaginase II, a potential antileukemic drug: Purification and characterization of the enzyme expressed in Pichia pastoris. Protein Expression and Purification, v.120, p. 118-125, 2016. | pt_BR |
| ISSN | 1046-5928 | pt_BR |
| URI | https://www.arca.fiocruz.br/handle/icict/14881 | |
| Language | eng | pt_BR |
| Publisher | Elsevier | pt_BR |
| Rights | restricted access | |
| Subject in Portuguese | Asparaginase | pt_BR |
| Subject in Portuguese | Drogas antitumorais | pt_BR |
| Subject in Portuguese | Purificação | pt_BR |
| Subject in Portuguese | Caracterização | pt_BR |
| Subject in Portuguese | Pichia pastoris | pt_BR |
| Title | Saccharomyces cerevisiae asparaginase II, a potential antileukemic drug: Purification and characterization of the enzyme expressed in Pichia pastoris | pt_BR |
| Type | Article | |
| DOI | 10.1016/j.pep.2015.12.012 | |
| Abstract | Asparaginase obtained from Escherichia coli and Erwinia chrysanthemi are used to treat acute lymphocytic leukaemia and non-Hodgkin's lymphoma. However, these agents cause severe adverse effects. Saccharomyces cerevisiae asparaginase II, encoded by the ASP3 gene, could be a potential candidate for the formulation of new drugs. This work aimed to purify and characterize the periplasmic asparaginase produced by a recombinant Pichia pastoris strain harbouring the ASP3 gene. The enzyme was purified to homogeneity with an activity recovery of 51.3%. The estimated molecular mass of the enzyme was 136 kDa (under native conditions) and 48.6 kDa and 44.6 kDa (under reducing conditions), suggesting an oligomeric structure. The recombinant asparaginase is apparently non-phosphorylated, and the major difference between the monomers seems to be their degree of glycosylation. The enzyme showed an isoelectric point of 4.5 and maximum activity at 46 °C and pH 7.2, retaining 92% of the activity at 37 °C. Circular dichroism and fluorescence analyses showed that the enzyme structure is predominantly α-helical with the contribution of β-sheet and that it remains stable up to 45 °C and in the pH range of 6-10. In vitro tests indicated that the recombinant asparaginase demonstrated antitumoural activity against K562 leukaemic cells. | pt_BR |
| Affilliation | Universidade Federal do Rio de Janeiro. Instituto de de Química. Departamento de Bioquímica. Laboratório de Tecnologia Enzimática. Rio de Janeiro, RJ, Brasil / Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Toxinologia. Rio de Janeiro, RJ, Brasil.. | pt_BR |
| Affilliation | Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Toxinologia. Rio de Janeiro, RJ, Brasil. | pt_BR |
| Affilliation | Universidade Federal do Rio de Janeiro. Instituto de de Química. Departamento de Bioquímica. Laboratório de Tecnologia Enzimática. Rio de Janeiro, RJ, Brasil. | pt_BR |
| Affilliation | Fundação Oswaldo Cruz. Biomanguinhos. Laboratório de Macromoléculas. Rio de Janeiro, RJ, Brasil. | pt_BR |
| Affilliation | Fundação Oswaldo Cruz. Farmanguinhos. Instituto de Tecnologia. Rio de Janeiro, RJ, Brasil. | pt_BR |
| Affilliation | Fundação Oswaldo Cruz. Biomanguinhos. Laboratório de Macromoléculas. Rio de Janeiro, RJ, Brasil. | pt_BR |
| Affilliation | Fundação Oswaldo Cruz. Farmanguinhos. Laboratório de Tecnologia. Rio de Janeiro, RJ, Brasil. | pt_BR |
| Affilliation | Universidade Federal do Rio de Janeiro. Instituto de de Química. Departamento de Bioquímica. Laboratório de Tecnologia Enzimática. Rio de Janeiro, RJ, Brasil. | pt_BR |
| Affilliation | Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Toxinologia. Rio de Janeiro, RJ, Brasil. | pt_BR |
| Subject | Asparaginase | pt_BR |
| Subject | Pichia pastoris | pt_BR |
| Subject | Antitumoural drug | pt_BR |
| Subject | Purification | pt_BR |
| Subject | Characterization | pt_BR |
| Embargo date | 2030-01-01 | |
