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Activities of Alkyl Hydroperoxide Reductase Subunits C1 and C2 of Vibrio parahaemolyticus against Different Peroxides.

Applied and environmental microbiology (2014-09-23)
Chun-Hui Chung, Tsung-Yong Ma, Shin-Yuan Fen, Hin-Chung Wong
ABSTRACT

Alkyl hydroperoxide reductase subunit C gene (ahpC) functions were characterized in Vibrio parahaemolyticus, a commonly occurring marine food-borne enteropathogenic bacterium. Two ahpC genes, ahpC1 (VPA1683) and ahpC2 (VP0580), encoded putative two-cysteine peroxiredoxins, which are highly similar to the homologous proteins of Vibrio vulnificus. The responses of deletion mutants of ahpC genes to various peroxides were compared with and without gene complementation and at different incubation temperatures. The growth of the ahpC1 mutant and ahpC1 ahpC2 double mutant in liquid medium was significantly inhibited by organic peroxides, cumene hydroperoxide and tert-butyl hydroperoxide. However, inhibition was higher at 12°C and 22°C than at 37°C. Inhibiting effects were prevented by the complementary ahpC1 gene. Inconsistent detoxification of H2O2 by ahpC genes was demonstrated in an agar medium but not in a liquid medium. Complementation with an ahpC2 gene partially restored the peroxidase effect in the double ahpC1 ahpC2 mutant at 22°C. This investigation reveals that ahpC1 is the chief peroxidase gene that acts against organic peroxides in V. parahaemolyticus and that the function of the ahpC genes is influenced by incubation temperature.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Luperox® TBH70X, tert-Butyl hydroperoxide solution, 70 wt. % in H2O
Sigma-Aldrich
tert-Butyl hydroperoxide solution, 5.0-6.0 M in decane
Sigma-Aldrich
Cumene, 98%
Sigma-Aldrich
tert-Butyl hydroperoxide solution, packed in FEP bottles, ~5.5 M in decane (over molecular sieve 4Å)
Sigma-Aldrich
tert-Butyl hydroperoxide solution, packed in FEP bottles, ~5.5 M in nonane (over molecular sieve 4 Å)
Sigma-Aldrich
tert-Butyl hydroperoxide solution, 5.0-6.0 M in nonane
Supelco
Cumene, Pharmaceutical Secondary Standard; Certified Reference Material
Supelco
Cumene, PESTANAL®