Biotechnological production of L-asparaginase (ASP1) of Saccharomyces cerevisiae in heterologous expression system Pichia pastoris

• Grant number: 13/14494-5
• Support Opportunities: Scholarships in Brazil - Master
• Start date: March 01, 2014
• End date: June 30, 2015
• Field of knowledge: Health Sciences - Pharmacy
• Principal Investigator: Adalberto Pessoa Junior
• Grantee: Bruna de Souza Divino
• Host Institution: Faculdade de Ciências Farmacêuticas (FCF). Universidade de São Paulo (USP). São Paulo , SP, Brazil

Abstract

Widely used in the treatment of acute lymphoblastic leukemia, L-Asparaginase is an enzyme that acts by decreasing the concentration of free asparagine in plasma. Therefore, prevents proliferation of malignant cells, since these cells, unlike healthy cells can not synthesize the amino acid asparagine to which require protein synthesis for not having the asparagine synthetase gene silencing due to a. The L-asparaginase currently used in Brazil is imported and bacteria, which creates problems with cost and supply, and cause allergies to some users. This encourages us to find ways of producing such an enzyme in a less costly for easy access and, if possible, with lower allergic potential. For this, we will study the biotechnological production of L-asparaginase in yeast heterologous expression system Pichia pastoris. The goal is to obtain a strain of P. pastoris expressing isoform of L-asparaginase extracellularly to facilitate purification process industry. Thus, there will be cloning the synthetic gene (ASP1) with codons optimized for expression in P. pastoris and optimization of production of L-asparaginase in shaker using an experimental design with the variables: pH, temperature, substrate concentration (glycerol), inoculum concentration, concentration of inducer and induction time. (AU)

Academic Publications

(References retrieved automatically from State of São Paulo Research Institutions)

DIVINO, Bruna de Souza. Biotechnological production of L-asparaginase (ASP1) of Saccharomyces cerevisiae in a heterologous expression system Pichia pastoris.. 2015. Master's Dissertation - Universidade de São Paulo (USP). Conjunto das Químicas (IQ e FCF) (CQ/DBDCQ) São Paulo.