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Appl Environ Microbiol ; 79(12): 3734-43, 2013 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-23563952

RESUMO

Alkyl hydroperoxide reductase subunit C (AhpC) is the catalytic subunit responsible for the detoxification of reactive oxygen species that form in bacterial cells or are derived from the host; thus, AhpC facilitates the survival of pathogenic bacteria under environmental stresses or during infection. This study investigates the role of AhpC in the induction and maintenance of a viable but nonculturable (VBNC) state in Vibrio parahaemolyticus. In this investigation, ahpC1 (VPA1683) and ahpC2 (VP0580) were identified in chromosomes II and I of this pathogen, respectively. Mutants with deletions of these two ahpC genes and their complementary strains were constructed from the parent strain KX-V231. The growth of these strains was monitored on tryptic soy agar-3% NaCl in the presence of the extrinsic peroxides H(2)O(2) and tert-butyl hydroperoxide (t-BOOH) at different incubation temperatures. The results revealed that both ahpC genes were protective against t-BOOH, while ahpC1 was protective against H(2)O(2). The protective function of ahpC2 at 4°C was higher than that of ahpC1. The times required to induce the VBNC state (4.7 weeks) at 4°C in a modified Morita mineral salt solution with 0.5% NaCl and then to maintain the VBNC state (4.7 weeks) in an ahpC2 mutant and an ahpC1 ahpC2 double mutant were significantly shorter than those for the parent strain (for induction, 6.2 weeks; for maintenance, 7.8 weeks) and the ahpC1 mutant (for induction, 6.0 weeks; for maintenance, 8.0 weeks) (P < 0.03). Complementation with an ahpC2 gene reversed the effects of the ahpC2 mutation in shortening the times for induction and maintenance of the VBNC state. This investigation identified the different functions of the two ahpC genes and confirmed the particular role of ahpC2 in the VBNC state of V. parahaemolyticus.


Assuntos
Genes Bacterianos/genética , Viabilidade Microbiana/efeitos dos fármacos , Peroxirredoxinas/metabolismo , Vibrio parahaemolyticus/enzimologia , Vibrio parahaemolyticus/crescimento & desenvolvimento , Análise de Variância , Primers do DNA/genética , Teste de Complementação Genética , Peróxido de Hidrogênio/metabolismo , Peróxido de Hidrogênio/toxicidade , Viabilidade Microbiana/genética , Peroxirredoxinas/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Deleção de Sequência/genética , Especificidade da Espécie , Temperatura , Vibrio parahaemolyticus/genética , terc-Butil Hidroperóxido/metabolismo , terc-Butil Hidroperóxido/toxicidade
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