Biotechnological production of L-asparaginase (ASP3.1) of Saccharomyces cerevisiae in heterologous expression system Pichia pastoris

Abstract

Biopharmaceuticals are mostly recombinant proteins produced by biotechnological tools. The L-asparaginase, an important biopharmaceutical, originating from bacteria, has been used as an effective therapeutic agent in the treatment of acute lymphoblastic leukemia (ALL) for over 40 years. However, recently this drug was removed from circulation in our country, causing much concern in the whole system of patient care onco-hematology, since the ALL develops rapidly and any delay in treatment can dramatically decrease the chances of healing. Thus, this project aims to generate a biopharmaceutical produced with national technology. For this, we will study the biotechnological production of yeast L-asparaginase in heterologous expression system Pichia pastoris. The objective is to obtain a strain of P. pastoris expressing isoform of L-asparaginase extracellularly to facilitate the purification industry process. For this will be made gene cloning synthetic (ASP3.1) with codons optimized for expression in P. pastoris and optimization of production of L-asparaginase in shaker using experimental design with the variables: pH, temperature, substrate concentration (glycerol), inoculum concentration, concentration of inducer and induction time.