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Doctoral Thesis
DOI
https://doi.org/10.11606/T.87.2017.tde-23022017-120752
Document
Author
Full name
Carlos Felipe Tasso Filietáz
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2016
Supervisor
Committee
Rocha, Adriana Rios Lopes (President)
Almeida, Fabio de
Lebrun, Ivo
Nakano, Eliana
Silva, Maria Cicera Pereira da
Title in Portuguese
Padrões de especificidade e expressão das lipases digestivas durante o desenvolvimento e o processo infeccioso no mosquito Aedes aegypti.
Keywords in Portuguese
Aedes aegypti
Enzimas digestivas
Lipase
Sistema digestório
Abstract in Portuguese
O mosquito Aedes aegypti é vetor de doenças como a febre amarela, dengue, chinkungunya e zika. O sistema digestório é responsável pela digestão e absorção de nutrientes, é também uma interface com o ambiente externo sendo a porta de entrada de organismos infecciosos. A presença de duas lipases digestivas foi confirmada por qPCR, uma na fase larval (L-Aa7051) e outra na fase adulta (L-Aa7055). Estas enzimas foram agrupadas na família das lipases neutras e apresentam alterações em resíduos envolvidos na especificidade, domínio tampa e alça β9. A L-Aa7055 recombinante foi expressa heterologamente em Escherichia coli na porção insolúvel, com atividade após a renaturação. Observamos que a expressão da lipase L-Aa7055 sofre uma redução de 30% na infecção Plasmodium gallinaceum, não sendo afetada pelo vírus dengue sorotipo 2 (DENV2). A digestão de lipídeos é importante na fase larval, com altos níveis de transcrito. Um estudo mais aprofundado ainda será necessário para compreender completamente o papel das lipases no processo infeccioso.
Title in English
Patterns of specificity and digestive lipases expression during the infectious process and development on the mosquito Aedes aegypti.
Keywords in English
Aedes aegypti
Digestive enzymes
Digestive system
Lipase
Abstract in English
The Aedes aegypti borne diseases yellow fever, dengue fever, chinkungunya and zica are important public healthy problems. The digestion and absorption of nutrients are performed in the digestive system, which is also an external environment interface that allows the infection by pathogenic microorganisms. The presence of two digestive lipases were identified by qPCR, L-Aa7051 in the larval phase and L-Aa7055 in the adult female. The lipase sequences were grouped in the neutral family, and exhibit alterations in residues involved in specificity, lid domain and β9 loop. The recombinant L-Aa7055 was expressed in the insoluble fraction, and show activity after a renaturation process. We notice that the expression levels of L-Aa7055 are reduced by 30% in the Plasmodium gallinaceum infection and were not affected by serotype 2 dengue virus (DENV2). The lipid digestion is important in the larval phase, with higher transcript levels. New studies will be necessary to the complete understanding of lipase contribution in the infectious process.
 
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Publishing Date
2017-02-23
 
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