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J Mol Microbiol Biotechnol ; 25(6): 412-22, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-26820469

RESUMO

BACKGROUND: Acetylation and efflux of carbohydrates during cellular metabolism is a well-described phenomenon associated with a detoxification process to prevent metabolic congestion. It is still unclear why cells discard important metabolizable energy sources in the form of acetylated compounds. METHODS: We describe the purification and characterization of an approximately 28-kDa intracellular carboxylesterase (YjfP) and the analysis of gene and protein expression by qRT-PCR and Western blot. RESULTS: qRT-PCR and Western blot, respectively, showed that yjfP is upregulated during the diauxic lag in cells growing with a mixture of glucose and lactose. The ß-galactosidase activity in the ΔyjfP strain was both delayed and half the magnitude of that of the wild-type strain. YjfP-hyperproducing strains displayed a long lag phase when cultured with glucose and then challenged to grow with lactose or galactose as the sole carbon source. CONCLUSION: Our results suggest that YjfP controls the intracellular concentration of acetyl sugars by redirecting them to the main metabolic circuits. Instead of detoxification, we propose that sugar acetylation is utilized by the cell for protection and to prevent the metabolism of a necessary minimal intracellular sugar pool. Those sugars can eventually be exported as a side effect of these mechanisms.


Assuntos
Carboxilesterase/genética , Carboxilesterase/metabolismo , Proteínas de Escherichia coli/genética , Proteínas de Escherichia coli/metabolismo , Escherichia coli/enzimologia , Escherichia coli/genética , Sequência de Bases , Western Blotting/métodos , Metabolismo dos Carboidratos , Carboxilesterase/química , Ativação Enzimática , Repressão Enzimática , Escherichia coli/crescimento & desenvolvimento , Proteínas de Escherichia coli/química , Galactose/metabolismo , Regulação Bacteriana da Expressão Gênica , Marcação de Genes , Glucose/metabolismo , Lactose/metabolismo , Reação em Cadeia da Polimerase em Tempo Real/métodos , Alinhamento de Sequência , Regulação para Cima , beta-Galactosidase/metabolismo
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