Bioprocess, purification and characterization of the peptidases produced by the fungi Eupenicillium javanicum and Myceliophthora thermophila and evaluation of the production of biologically active peptides

Author(s)

Youssef Ali Abou Hamin Neto

• Document type: Doctoral Thesis
• Press: Ribeirão Preto.
• Institution: Universidade de São Paulo (USP). Faculdade de Ciências Farmacêuticas de Ribeirão Preto (PCARP/BC)
• Defense date: 2017-03-29

Examining board members

Hamilton Cabral; Vitor Marcel Faça; Luis Henrique Souza Guimarães; Ana Patricia Yatsuda Natsui; Arthur Henrique Cavalcante de Oliveira

• Advisor: Hamilton Cabral

Abstract

The cancer is a disease responsible to the higher levels of mortality and morbity around the world. This disease is related to many factors, as environment, nutrition, genetic, virus infection and life style.The complexity of this disease increases the research of new treatments. Currently, the medicines damage both, normal and cancer cells, it promotes side effects and toxicity. Some molecules present specificity and bind to proteins from membranes of cancer cells, some examples are peptides. Peptides can be obtained from proteins by hydrolisys with microrbial proteases. The aim of this study was to produce and purify two proteases from fungi, Eupenicillium javanicum and Myceliophthora thermophila. Biochemical characterization and kinetic assays of these enzymes were evaluated. And then, the proteases were applied on obtention of peptides from milk, egg and gelatin proteins, and the antioxidant (in vitro) and antitumoral activity (in vitro e in vivo) of these peptides were evaluated. The purified enzymes belong to metalo (E. javanicum) and serine (M. thermophila) protease classes, the kinetic assay showed higher catalytic efficiency with tyrosine at P\'1 position and alanine at P\'2 position of polypeptide chain of substrate, to metalo and serine protease, respectively. The peptides from casein presented capacity to scavenge free radicals and to quelate iron. Peptides derived from whey and gelatin inhibited the grown of oral cancer and melanoma cells (in vitro). Peptides obtained from whey decreased the tumor volume of immunosupressed mice. Proteomic assays of these tumors, when compared treatment and control group, showed that peptides can be envolved in some apoptosys pathway and it influences the tumor volume, cosequently, it is decreased on treatment with peptides derived from milk whey (AU)

• FAPESP's process: 12/18278-2 - Bioprocess, purification and characterization of peptidases produced by fungi Eupenicillium javanicum and Myceliophthora sp. and evaluating the production of biologically active peptides
• Grantee: Youssef Ali Abou Hamin Neto
• Support Opportunities: Scholarships in Brazil - Doctorate