Biotechnological production of L- asparaginase ( ASP3 ) of Saccharomyces cerevisiae in a heterologous expression system Pichia pastoris

Author(s)

Rafaela Coelho Correia

• Document type: Master's Dissertation
• Press: São Paulo.
• Institution: Universidade de São Paulo (USP). Conjunto das Químicas (IQ e FCF) (CQ/DBDCQ)
• Defense date: 2015-11-26

Examining board members

Adalberto Pessoa Junior; Valéria de Carvalho Santos Ebinuma; Ricardo Pinheiro de Souza Oliveira

• Advisor: Adalberto Pessoa Junior; Gisele Monteiro de Souza

Abstract

Acute lymphoblastic leukemia (ALL) is considered a serious disease of white blood cells, is more common and more aggressive in children and adolescents. Treatment for ALL has advanced due to studies for drug optimization already used in chemotherapy. Among these drugs are the enzymes L-asparaginases (ASPases) which act by reducing the concentration of L-asparagine (Asn) in the bloodstream, preventing the proliferation of cancer cells, since these can not synthesize appropriate amounts of this amino acid. However, the drug to be derived from a prokaryote causes severe allergic reactions to the user, in order to decrease the immunogenicity of the chemotherapy, it is important to generate a biopharmaceutical derived from a eukaryote. In this context, we obtained the recombinant Pichia pastoris responsible for producing the enzyme ASPase intermembrane, coming from the ASP3 gene of Saccharomyces cerevisiae. Through the experimental design, it was possible to have a 5-fold increase in activity obtained at the initial condition. The Mut + clone achieved their best activity of 8.6 U/g cell under the following conditions: 20 °C, initial pH 6 and 1.5% of inducer concentration. (AU)

• FAPESP's process: 13/13881-5 - Biotechnological production of L-asparaginase (ASP3.1) of Saccharomyces cerevisiae in heterologous expression system Pichia pastoris
• Grantee: Rafaela Coelho Correia
• Support Opportunities: Scholarships in Brazil - Master