Data/hora: 06/06/2024 02:42:28

Provedor de dados: | 74
País: | Brazil
Título: | Carbohydrate metabolism of Xylella fastidiosa: Detection of glycolytic and pentose phosphate pathway enzymes and cloning and expression of the enolase gene
Autores: | Facincani,Agda Paula; Ferro,Jesus Aparecido; Pizauro Jr.,João Martins; Pereira Jr.,Haroldo Alves; Lemos,Eliana Gertrudes de Macedo; Prado,Alessandro Luis do; Ferro,Maria Inês T.
Data: | 2003-01-01
Ano: | 2003
Palavras-chave: | Enolase; Entner-Doudoroff; Enzyme; Glycolysis; Xylella fastidiosa.
Resumo: | The objective of this work was to assess the functionality of the glycolytic pathways in the bacterium Xylella fastidiosa. To this effect, the enzymes phosphoglucose isomerase, aldolase, glyceraldehyde-3-phosphate dehydrogenase and pyruvate kinase of the glycolytic pathway, and glucose 6-phosphate dehydrogenase of the Entner-Doudoroff pathway were studied, followed by cloning and expression studies of the enolase gene and determination of its activity. These studies showed that X. fastidiosa does not use the glycolytic pathway to metabolize carbohydrates, which explains the increased duplication time of this phytopatogen. Recombinant enolase was expressed as inclusion bodies and solubilized with urea (most efficient extractor), Triton X-100, and TCA. Enolase extracted from X. fastidiosa and from chicken muscle and liver is irreversibly inactivated by urea. The purification of enolase was partial and resulted in a low yield. No enzymatic activity was detected for either recombinant and native enolases, aldolase, and glyceraldehyde-3-phosphate dehydrogenase, suggesting that X. fastidiosa uses the Entner-Doudoroff pathway to produce pyruvate. Evidence is presented supporting the idea that the regulation of genes and the presence of isoforms with regulation patterns might make it difficult to understand the metabolism of carbohydrates in X. fastidiosa.
Tipo: | Info:eu-repo/semantics/article
Idioma: | Inglês
Identificador: | http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572003000200015
Editor: | Sociedade Brasileira de Genética
Relação: | 10.1590/S1415-47572003000200015
Formato: | text/html
Fonte: | Genetics and Molecular Biology  v.26 n.2 2003
Fonte: | info:eu-repo/semantics/openAccess