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Effects of protein-aqueous medium interactions on structural modifications of L-asparaginase II of Escherichia coli

Grant number: 13/24024-6
Support Opportunities:Scholarships in Brazil - Doctorate
Effective date (Start): March 01, 2014
Effective date (End): August 31, 2017
Field of knowledge:Biological Sciences - Biochemistry - Biochemistry of Microorganisms
Principal Investigator:Gisele Monteiro
Grantee:Samarina Rodrigues Wlodarczyk
Host Institution: Faculdade de Ciências Farmacêuticas (FCF). Universidade de São Paulo (USP). São Paulo , SP, Brazil


The L- Asparaginase is an enzyme applied to the treatment of several types of cancer, mainly, Acute Lymphoblastic Leukemia. It is produced by the microorganisms Escherichia coli and Erwinia chrysanthemi by the induction of the native protein production or by the expression of the recombinant protein. The enzymes catalyses the conversion of the L- Asparagine to L- Aspartate and ammonia, depriving the tumor cell of an essential amino acid for its growing metabolism. Commercially, the enzymes available in the market for the treatment are produced through a process that needs to be improved. The formation of protein aggregates is one of the main problems that affect the formulation of L- Asparaginase. These may compromise the enzyme purification, cause the loss of the protein activity and even increase the immune response to the biotherapeutic drug. This project will have the main objective of analyze the behavior of the L- Asparaginase protein in several Aqueous Two-Phase Systems (ATPS), similar to the organic medium, and factors that are responsible for its aggregation. The dipolarity/polarizability, acidity and basicity of L- Asparaginase will be estimated by solvatochromic studies and the generation of descriptors to this enzyme. The effects of site-directed mutagenesis will also be evaluated in the ATPS. The results may be helpful to the comprehension of biological functions and the transport of proteins, and improve the development of biotherapeutic drugs. (AU)

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Scientific publications
(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)
WLODARCZYK, SAMARINA R.; CUSTODIO, DEBORA; PESSOA JR, ADALBERTO; MONTEIRO, GISELE. Influence and effect of osmolytes in biopharmaceutical formulations. EUROPEAN JOURNAL OF PHARMACEUTICS AND BIOPHARMACEUTICS, v. 131, p. 92-98, . (15/24463-5, 13/08617-7, 13/24024-6, 15/07749-2)
WLODARCZYK, SAMARINA R.; COSTA-SILVA, TALES A.; PESSOA-JR, ADALBERTO; MADEIRA, PEDRO; MONTEIRO, GISELE. Effect of osmolytes on the activity of anti-cancer enzyme L-Asparaginase II from Erwinia chrysanthemi. Process Biochemistry, v. 81, p. 123-131, . (13/24024-6, 15/07749-2)

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