| Full text | |
| Author(s): |
Costa, Iris Munhoz
;
Schultz, Leonardo
;
Bianchi Pedra, Beatriz de Araujo
;
Moreira Leite, Mariana Silva
;
Farsky, Sandra H. P.
;
de Oliveira, Marcos Antonio
;
Pessoa, Adalberto
;
Monteiro, Gisele
Total Authors: 8
|
| Document type: | Journal article |
| Source: | SCIENTIFIC REPORTS; v. 6, NOV 8 2016. |
| Web of Science Citations: | 13 |
| Abstract | |
L-asparaginase (L-ASNase) (EC 3.5.1.1) is an important enzyme for the treatment of acute lymphoblastic leukaemia. Currently, the enzyme is obtained from bacteria, Escherichia coli and Erwinia chrysanthemi. The bacterial enzymes family is subdivided in type I and type II; nevertheless, only type II have been employed in therapeutic proceedings. However, bacterial enzymes are susceptible to induce immune responses, leading to a high incidence of adverse effects compromising the effectiveness of the treatment. Therefore, alternative sources of L-ASNase may be useful to reduce toxicity and enhance efficacy. The yeast Saccharomyces cerevisiae has the ASP1 gene responsible for encoding L-asparaginase 1 (ScASNase1), an enzyme predicted as type II, like bacterial therapeutic isoforms, but it has been poorly studied. Here we characterised ScASNase1 using a recombinant enzyme purified by affinity chromatography. ScASNase1 has specific activity of 196.2 U/mg and allosteric behaviour, like type I enzymes, but with a low K-0.5 = 75 mu M like therapeutic type II. We showed through site-directed mutagenesis that the T64-Y78-T141-K215 residues are involved in catalysis. Furthermore, ScASNase1 showed cytotoxicity for the MOLT-4 leukemic cell lineage. Our data show that ScASNase1 has characteristics described for the two subfamilies of l-asparaginase, types I and II, and may have promising antineoplastic properties. (AU) | |
| FAPESP's process: | 13/16685-2 - Production, kinetic characterization and engineering of Asparaginase 1 protein of the Saccharomyces cerevisiae for evaluate its use as biopharmaceutical. |
| Grantee: | Iris Munhoz Costa |
| Support Opportunities: | Scholarships in Brazil - Master |
| FAPESP's process: | 13/08617-7 - Production of extracellular L-asparaginase: from bioprospecting to the engineering of an antileukemic biopharmaceutical |
| Grantee: | Adalberto Pessoa Junior |
| Support Opportunities: | Research Projects - Thematic Grants |
| FAPESP's process: | 15/07749-2 - Protein engineering and comparison of microbial expression systems of the biopharmaceutical L-asparaginase. |
| Grantee: | Gisele Monteiro |
| Support Opportunities: | Regular Research Grants |