Analysis of secretome from Phanerochaete chrysosporium and post-translational modifications of some proteins during the lapachol biotransformation process by mass spectrometry

• Grant number: 16/12516-0
• Support Opportunities: Scholarships abroad - Research Internship - Doctorate
• Start date: October 01, 2016
• End date: July 31, 2017
• Field of knowledge: Biological Sciences - Biochemistry - Biochemistry of Microorganisms
• Principal Investigator: Hamilton Cabral
• Grantee: Luciana Barbosa Coitinho
• Supervisor: Loic Quinton
• Host Institution: Faculdade de Ciências Farmacêuticas de Ribeirão Preto (FCFRP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil
• Institution abroad: Université de Liège (ULg), Belgium
• Associated to the scholarship: 14/01497-9 - Biotransformation of lapachol by filamentous fungi: proteomic analysis for identification to biotransformation route, BP.DR

Abstract

Lapachol is a naphtoquinone extracted from plants of Bignoniaceae family with diverse biological activities such as anti-cancer, anti-inflammatory and others, being, therefore, of great interest to pharmaceutical industry. This molecule has been extensively studied in search of new bioactive analogs and one way to obtain these derivatives is by biotransformations studies. Fungi have been widely used in biotransformation studies in order to obtain new derivatives of natural or synthetic products, as well as a model to study metabolism of drugs. However, the metabolic pathways used by fungi to modify those bioactive molecules is poorly studied. These information are of great biotechnological relevance, since enzymes of industrial interest could be identified. Proteomic studies of the fungi during the biotransformation process can identify how these microorganisms transform the molecules and consequently a biotransformation route can be determined. Previous studies developed in Brazil showed that the fungus Phanerochaete chrysosporium is capable of biotransform the lapachol into two major products, and one of them is a novel compound in the literature. It was analyzed both the intracellular and extracellular proteome of P. chrysosporium. Results obtained by two-dimensional electrophoresis (2DE) followed by MALDI TOF/TOF, showed that the fungus modifies the production of intracellular proteins in the presence of lapachol, producing mainly oxidative enzymes. On the other hand, the profile of extracellular proteome (secretome) was analyzed by LC-MS/MS, however this technique could not detect any protein. Besides that, the amount of protein produced in the secretome is very small. So, a more sensitive spectrometry mass equipment than the one we have available here in our college could help to obtain these fundamental data to complete our study. In this context, the objective of this study is to analyze the secretome of P. chrysosporium by mass spectrometry proposing a biotransformation route, and analyze the post-translational modifications of exclusive proteins involved in the lapachol biotransformation process. The Mass Spectrometry Laboratory at Université de Liège has instruments suitable to applications requiring sub-ppm mass determination or for very complex mixtures containing compounds with very close masses such as secretoma from P. chrysosporium. The realization of this internship in an internationally recognized research group in mass spectrometry field will contribute to the formation of the student in mass spectrometry area, allowing not only the improvement of the impact of her publications but also providing a broad technological improvement of the research group in Brazil. The internship will also contribute to the establishment of a permanent cooperation between the two institutions.

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)

AMORIM, FERNANDA GOBBI; COITINHO, LUCIANA BARBOSA; DIAS, ANANDA TISSIANEL; FREITAS FRIQUES, ANDREIA GOMES; MONTEIRO, BRENNA LEPAUS; DIAS DE REZENDE, LUCAS CUNHA; COSTA PEREIRA, THIAGO DE MELO; CAMPAGNARO, BIANCA PRANDI; DE PAUW, EDWIN; VASQUEZ, ELISARDO CORRAL; et al. Identification of new bioactive peptides from Kefir milk through proteopeptidomics: Bioprospection of antihypertensive molecules. Food Chemistry, v. 282, p. 109-119, JUN 2019. (14/01497-9, 16/12516-0)