RESUMO
An ongoing outbreak of multidrug-resistant Salmonella enterica serovar Anatum began in Taiwan in 2015. Pork and poultry were identified as vehicles for transmission. Contaminated meat contributed to the high rate of infections among children. Nearly identical Salmonella Anatum strains have been identified in the United Kingdom, the United States, and the Philippines.
Assuntos
Antibacterianos , Salmonella enterica , Salmonella , Animais , Antibacterianos/farmacologia , Criança , Surtos de Doenças , Farmacorresistência Bacteriana Múltipla/efeitos dos fármacos , Humanos , Carne , Testes de Sensibilidade Microbiana , Filipinas , Salmonella/efeitos dos fármacos , Salmonella/genética , Salmonella/isolamento & purificação , Salmonella enterica/efeitos dos fármacos , Salmonella enterica/genética , Taiwan/epidemiologia , Reino Unido , Estados UnidosRESUMO
A gene that codes for a novel intracellular poly(3-hydroxybutyrate) (PHB) depolymerase, designated PhaZ1, has been identified in the genome of Bacillus megaterium. A native PHB (nPHB) granule-binding assay showed that purified soluble PhaZ1 had strong affinity for nPHB granules. Turbidimetric analyses revealed that PhaZ1 could rapidly degrade nPHB granules in vitro without the need for protease pretreatment of the granules to remove surface proteins. Notably, almost all the final hydrolytic products produced from the in vitro degradation of nPHB granules by PhaZ1 were 3-hydroxybutyric acid (3HB) monomers. Unexpectedly, PhaZ1 could also hydrolyze denatured semicrystalline PHB, with the generation of 3HB monomers. The disruption of the phaZ1 gene significantly affected intracellular PHB mobilization during the PHB-degrading stage in B. megaterium, as demonstrated by transmission electron microscopy and the measurement of the PHB content. These results indicate that PhaZ1 is functional in intracellular PHB mobilization in vivo. Some of these features, which are in striking contrast with those of other known nPHB granule-degrading PhaZs, may provide an advantage for B. megaterium PhaZ1 in fermentative production of the biotechnologically valuable chiral compound (R)-3HB.
Assuntos
Bacillus megaterium/enzimologia , Hidrolases de Éster Carboxílico , Regulação Bacteriana da Expressão Gênica , Hidroxibutiratos/metabolismo , Poliésteres/metabolismo , Sequência de Aminoácidos , Bacillus megaterium/genética , Bacillus megaterium/crescimento & desenvolvimento , Bacillus megaterium/ultraestrutura , Proteínas de Bactérias/química , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Hidrolases de Éster Carboxílico/química , Hidrolases de Éster Carboxílico/genética , Hidrolases de Éster Carboxílico/metabolismo , DNA Bacteriano/genética , Hidroxibutiratos/química , Microscopia Eletrônica de Transmissão , Dados de Sequência Molecular , Mutação , Plasmídeos , Poliésteres/química , Alinhamento de Sequência , Análise de Sequência de DNARESUMO
Bacillus thuringiensis is a gram-positive spore-forming bacterium that can accumulate poly(3-hydroxybutyrate) (PHB) as a carbon and energy storage substance in response to nutritional stress. The regulatory mechanism for PHB biosynthesis in B. thuringiensis and diverse Bacillus species is still poorly understood. We now report that disruption of the sigH gene or the gene encoding the master sporulation transcription factor Spo0A severely impaired PHB accumulation in B. thuringiensis. Complementation of the spo0A mutation with the spo0A gene restored PHB accumulation. We have found that the requirement of Spo0A for PHB accumulation is independent of the transition state regulator AbrB and of loss of sporulation ability. We also show that Spo0A is required for the expression of three genes involved in PHB biosynthesis. These findings have uncovered a new role of Spo0A in the regulation of stationary-phase-associated cellular events.