Orientador: Anita Jocelyne Marsaioli / Tese (doutorado) - Universidade Estadual de Campinas, Instituto de Química / Made available in DSpace on 2018-08-25T12:48:43Z (GMT). No. of bitstreams: 1 Angolini_CelioFernandoFigueiredo_D.pdf: 5315143 bytes, checksum: 8dd92777c4d60094caaa95a789eb6ae2 (MD5) Previous issue date: 2014 / Resumo: Neste trabalho avaliou-se a ação de micro-organismos em amostras de petróleo naturalmente biodegradadas e em ensaios de biodegradação realizados em laboratório, através da otimização de um protocolo que utiliza metodologias rápidas de análise da fração polar por ESI-MS. Assim sendo foi possível aferir a composição molecular dos constituintes das classes de compostos oxigenados (O, O2, O3 e O4) e nitrogenados (N, NO, NO2, NO3) presentes no petróleo e correlacioná-las ao fenômeno de biodegradação, e ambiente deposicional em óleos da Bacia de Campo e da Bacia Potiguar, respectivamente. As informações geradas nesse estudo contribuíram para o entendimento da origem dos biomarcadores ácidos através da degradação dos respectivos hidrocarbonetos. Complementando o escopo principal do trabalho foram caracterizados dos micro-organismos do petróleo por técnicas de espectrometria de massas, mostrando a importância do desenvolvimento desta técnica com construções de bibliotecas de referência e elaboração de softwares mais elaborados. Finalizando foi realizada a síntese de um biomarcador trideuterado com ampla utilização em estudos de petróleo / Abstract: In this work we were able to evaluate the action of microorganisms in naturally biodegraded samples and in biodegraded laboratory assays, by optimizing a protocol that uses fast methods of analysis of the polar fraction by ESI-MS. Which was possible to determine the molecular composition of the compounds for oxygen (O, O2, O3 and O4) and nitrogen classes (N, NO, NO2, NO3) present in petroleum. Allowing to correlate those classes with the biodegradation phenomenon, and the depositional environment for the different studied petroleum samples. Even more information is generated about the origin of biomarkers acids through degradation of the respective hydrocarbons. In addition to the main scope of work, we characterize the petroleum microorganisms by mass spectrometry techniques, showing the importance of developing this MO identification technique by the construction of reference libraries and development of more sophisticated software. Also we synthesized a trideuterated biomarker widely used in studies with petroleum / Doutorado / Quimica Organica / Doutor em Ciências

Antibiotic resistance is increasing rapidly world wide. Resistance determinants have evolved long before antibiotics were used. Though horizontal gene transfer and mutation play a major role in the dissemination of antibiotic resistance genes their evolutionary origins remain obscure. A model system was used to investigate how they might arise in the first case. Plasmid borne erythromycin resistant clones were selected through marker rescue from genomic libraries of DNA from Gram positive organisms, maintained in E. coli. EryR determinants were recovered from nine libraries of 23 screened. Clone pMP1 (DNA from Mycobacterium parafortuitum) was the most resistant, with an MIC of 400 ìg/ml for erythromycin and 12 ìg/ml for azithromycin. Antibiotic resistance was not expressed in Rhodococcus erythropolis. Restriction maps were constructed for clones pMP1 and pMCX (DNA from Mycobacterium avium). Clone pMP1 was sub-cloned and the fragment carrying the EryR determinant (~2.4 kb) was sequenced. Analysis revealed 2 open reading frames (ORF1 and ORF2). ORF1 showed highest similarity to an FixB/FixA proteins and ORF2 showed similarity to a methyl transferase. Key words: antibiotic resistance,erythromycin, Gram positive DNA / Prof. E.R. Dabbs