Cloning, expression and mapping of non-classical peptidase subsites: biotechnological application in obtaining bioactive peptides

• Grant number: 20/14426-3
• Support Opportunities: Regular Research Grants
• Start date: December 01, 2021
• End date: May 31, 2024
• Field of knowledge: Biological Sciences - Biochemistry - Enzymology
• Principal Investigator: Hamilton Cabral
• Grantee: Hamilton Cabral
• Host Institution: Faculdade de Ciências Farmacêuticas de Ribeirão Preto (FCFRP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil
• Associated researchers: Andréia Machado Leopoldino

Abstract

The prospects for the use of industrial microbial enzymes have increased considerably in the 21st century, with an increasing trend as the new enzymes present significant potential to meet the needs of several industrial sectors. Enzymes of microbial origin, besides being robust, have significant potential in waste management and, consequently, in the development of a safer environment. The enzyme market is expected to reach a value of US $ 14.7 billion in 2025, registering an annual growth rate composed of 6.7 in terms of values. Microbial peptidases constitute the main group of enzymes used industrially, responsible for about 2/3 of the total of commercialized enzymes. The study of fungal peptidase is a relevant topic worldwide, as it involves physiological studies that help to understand how some pathogenic fungi interact with hosts, in the field of biotechnology, with the use of enzymes from filamentous fungi in several industrial sectors. Currently, fungal enzymes are the most used, for example, in the food, pharmaceutical, biofuel and other industries. The importance in determining the specificity of peptidases, that is, the mapping of subsites, helps in the rational design of inhibitors, makes it possible to determine the cleavage in proteins and to obtain peptides with biological activity. Currently, peptides are considered as a new class of drugs that is emerging, in 2017 the global market for bioactive proteins and peptides was valued at US $ 88.3 billion, with a compound annual growth rate from 2020 to 2027 from 8 ,2%. The present project aims to clone the genes that encode threonine and omega peptidases of filamentous fungi in the expression system in Pichia pastoris. Due to the need for greater knowledge of this subclass of hydrolases, in addition to the potential for biotechnological application, as it is a project involving the cloning and expression of new enzymes that have wide applications in several areas, this project has great potential for new applications for patents, in addition to the scientific articles that will be produced with the results obtained, in addition to the possibility of discovering peptides with biological activity of medical importance. (AU)

Scientific publications

(4)

(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)

PESSELA, BENEVIDES COSTA; OKAMOTO, DEBORA NOMA; CABRAL, HAMILTON. Editorial: Microbial proteases: Biochemical studies, immobilization and biotechnological application. FRONTIERS IN MICROBIOLOGY, v. 14, p. 2-pg., 2023-01-27. (16/20385-2, 20/14426-3)

YOUSSEF ALI ABOU HAMIN NETO; NATHÁLIA GONSALES DA ROSA GARZON; LUCIANA BARBOSA COITINHO; LAYS MARTIN SOBRAL; ANDRÉIA MACHADO LEOPOLDINO; THAÍS REGIANI CATALDI; CARLOS ALBERTO LABATE; HAMILTON CABRAL. Fungal metalloprotease generate whey-derived peptides that may be involved in apoptosis in B16F10 melanoma cells. FOOD SCIENCE AND TECHNOLOGY, v. 42, 2022-07-26. (12/18278-2, 20/14426-3, 12/24703-8)

PEDEZZI, RAFAEL; EVANGELISTA, DANILO ELTON; GARZON, NATHALIA GONSALES DA ROSA; SIMOES, FLAVIO ANTONIO DE OLIVEIRA; DE OLIVEIRA, ARTHUR HENRIQUE CAVALCANTE; POLIKARPOV, IGOR; CABRAL, HAMILTON. Biochemical and biophysical properties of a recombinant serine peptidase from Purpureocillium lilacinum. Biophysical Chemistry, v. 296, p. 9-pg., 2023-02-22. (20/14426-3, 15/16084-4)

FURLAN, JOAO PEDRO RUEDA; RIBEIRO, TERESA GONSALVES; NOVAIS, ANGELA; CABRAL, HAMILTON; PEIXE, LUISA; STEHLING, ELIANA GUEDES. Citrobacter meridianamericanus sp. nov., isolated from a soil sample. INTERNATIONAL JOURNAL OF SYSTEMATIC AND EVOLUTIONARY MICROBIOLOGY, v. 73, n. 3, p. 5-pg., 2023-01-01. (21/01655-7, 18/01890-3, 20/14426-3)