Advanced search
Start date
Betweenand


Applications of Nuclear Magnetic Resonance in metabolomic studies of biological process

Full text
Author(s):
Paulo Falco Cobra
Total Authors: 1
Document type: Doctoral Thesis
Press: São Carlos.
Institution: Universidade de São Paulo (USP). Instituto de Química de São Carlos (IQSC/BT)
Defense date:
Examining board members:
Luiz Alberto Colnago; Lucineia Vizzotto Marconcini; Tatiana Santana Ribeiro; Otavio Henrique Thiemann
Advisor: Luiz Alberto Colnago
Abstract

NMR is one of the main techniques used in metabolic profile studies because it is a quantitative, highly reproducible and non-selective technique. Thus, the aim of this thesis was the application of NMR in metabolic studies of different biological processes. The first application was the analysis of the metabolomic and metabonomic profile of the causative agents of leishmaniasis. Cell extracts of the species Leishmania major and Leishmania donovani and extracts of these species grown with miltefosine and paromomycin drugs were analyzed. 1H spectra were analyzed by ChenoMX software and 2D spectra by TopSpin software and HMDB and BMRB databases. About 50 metabolites were identified, such as adenine, arginine, glutamine, glutamate, mannose, isoleucine, leucine and tyrosine. Chemometric techniques were used to investigate the metabolites responsible for the differentiation of the two species and between cultures with and without drugs. The second study was with methylglyoxal molecule. The presence of methylglyoxal in cheese contributes to browning and deterioration of flavor, even at low temperatures. For this reason, products of the addition of methylglyoxal in Parmesan cheese extract and the behavior of different species of lactobacilli in the presence and absence of methylglyoxal in Parmesan cheese extract were studied. The results allowed a better understanding of the reactions involved between this molecule and the components of the cheese, which will help find alternatives to solve the problem and increase the shelf life of the cheese. The third study was about the production of ethanol from lignocellulosic biomass. The objective of this study was to genetically modify a lactobacillus species to produce ethanol and analyze the post-fermentation thin stillage. NMR spectra showed that the modified bacteria are efficient in production of ethanol and that two sugars were erroneously being identified by chromatography as maltose and maltotriose when they actually were trehalose and arabinose. These studies showed part of the numerous applications that NMR has in metabolomics and that there are several statistical tools available to assist in metabolomic studies. (AU)

FAPESP's process: 12/12847-5 - Analysis of leishmaniasis metabolic profile by NMR
Grantee:Paulo Falco Cobra
Support Opportunities: Scholarships in Brazil - Doctorate