Skopuleriopsis koningii enzyme prospection using proteome: Recombinant production of enzimes with biotecnological potential and evaluation of Antifungal

• Grant number: 15/16084-4
• Support Opportunities: Scholarships in Brazil - Doctorate
• Start date: August 01, 2016
• End date: July 31, 2019
• Field of knowledge: Biological Sciences - Biochemistry - Biochemistry of Microorganisms
• Principal Investigator: Hamilton Cabral
• Grantee: Rafael Pedezzi
• Host Institution: Faculdade de Ciências Farmacêuticas de Ribeirão Preto (FCFRP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil

Abstract

The biotechnological industry has growing around the world, including Brazil. However, research and development national funding is under the satisfactory level. The production of enzymes is an important biotechnological branch given that represents high economic gains and is in expansion. Microorganisms are the most applied in enzyme production, given its low cust of manipulation and high yield of the process. Peptidases are the most produced enzymes and show a large range of applications. Considering the Skopulariopsis koningii enzyme prospection, we propose a proteomic study based on differential expression inducing processes. The organism will be induced in different medium composition. The induction will be verified with 1D and 2D electrophoresis gel. The selected proteins will be sequenced in mass spectrometer to identification. We have two aims: collagenolitic peptidases prospection and prospection of proteins related to antifungal response. We also intend to recombinantly produce these enzymes in the yeast Pichia pastoris, under methanol induction. After the purification, enzymatic characterizations will be done, as substrate specificity, kinetic parameters, temperature and pH influence and other relevant tests. The enzymes might show activity close to the endogens. We expect find collagenases with potential to application in pharmaceutical industry. Moreover, the description of metabolic pathway response to antifungal resistance can be enlightening and open a way to depth studies.

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)

PEDEZZI, RAFAEL; DE BIAGI JUNIOR, CARLOS ALBERTO OLIVEIRA; DE FREITAS, MARCELA CRISTINA CORREA; DA ROSA-GARZON, NATHALIA GONSALES; CABRAL, HAMILTON. Transcriptomic studies on Purpureocillium lilacinum reveal molecular mechanisms of response to fluconazole and itraconazole. Brazilian Journal of Microbiology, v. 52, n. 2, MAR 2021. (15/16084-4, 16/20385-2)

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)

PEDEZZI, RAFAEL; EVANGELISTA, DANILO E.; POLIKARPOV, IGOR; CABRAL, HAMILTON. Recombinant production and structural studies of a serine peptidase from Scopulariopsis koningii. FASEB JOURNAL, v. 33, p. 2-pg., 2019-04-01. (16/20385-2, 15/16084-4)

Academic Publications

(References retrieved automatically from State of São Paulo Research Institutions)

PEDEZZI, Rafael. Prospecting of enzymes in Purpureocillium lilacinum using proteomic technique: recombinant production of enzymes with biotechnological potential and evaluation of antifungals. 2019. Doctoral Thesis - Universidade de São Paulo (USP). Faculdade de Ciências Farmacêuticas de Ribeirão Preto (PCARP/BC) Ribeirão Preto.