Production and purification of recombinant proteins of pharmaceutical interest

• Grant number: 14/50237-0
• Support Opportunities: Regular Research Grants
• Start date: August 01, 2014
• End date: March 31, 2016
• Field of knowledge: Biological Sciences - Microbiology - Biology and Physiology of Microorganisms
• Agreement: Universidad de la Frontera
• Principal Investigator: Adalberto Pessoa Junior
• Grantee: Adalberto Pessoa Junior
• Principal researcher abroad: Jorge Farias Avendaño
• Institution abroad: Universidad de La Frontera (UFRO), Chile
• Host Institution: Faculdade de Ciências Farmacêuticas (FCF). Universidade de São Paulo (USP). São Paulo , SP, Brazil
• Associated research grant: 13/08617-7 - Production of extracellular L-asparaginase: from bioprospecting to the engineering of an antileukemic biopharmaceutical, AP.TEM

Abstract

There is a strong global tendency to find alternative ways to produce pharmaceutical active principies from biotechnology processo In this scenario, Brazil shows a small expression in both research and production. Worsening the situation, international suppliers of biopharmaceuticals to Brazil are losing interest in the market and discontinuing production, especially those related to onco-hematologic treatment. In this context, this project proposes the union of different scientific and technological skills to achieve a viable industrial process to biotechnologically produce L-Asparaginase, a biopharmaceutical broadly used in the treatment of leukemia. This project will approach two main areas: optimization of endogenous and heterologous enzyme production, with bioprospecting groups of fungi from diverse biomes; and the protein rational engineering using L-Asparaginase from S. cerevisiae as scaffold for comparative studies with the bacterial isoforms currently employed in the therapy. Furthermore, crystallographic structures will be determined to guide the site-directed mutations and random mutations produced by protein synthetic evolution will be evaluated. To improve stability, bioavailability, toxicity and allergenic aspects, many nanotechnology approaches, such as pegylation and nanoencapsulation in polimerosomes, will be investigated. As a result, we expect this project can bring a national biopharmaceutical to the market. Both groups are expecting academic gains in new knowledge according to the expertise of the researchers involved in this proposal. For example, the USP group will receive researchers with experience in enzyme characterization, and metabolic engineering of enzymes, as well as the UFRO group will receive experts in fermentation technology, nanotechnology, liquid liquid extraction of proteins, and molecular biology of yeasts. (AU)