title: Factors contributing to hydrogen peroxide resistance in Streptococcus pneumoniae include pyruvate oxidase (SpxB) and avoidance of the toxic effects of the fenton reaction.
abstract: Aerobic growth of Streptococcus pneumoniae results in production of amounts of hydrogen peroxide (H(2)O(2)) that may exceed 1 mM in the surrounding media. H(2)O(2) production by S. pneumoniae has been shown to kill or inhibit the growth of other respiratory tract flora, as well as to have cytotoxic effects on host cells and tissue. The mechanisms allowing S. pneumoniae, a catalase-deficient species, to survive endogenously generated concentrations of H(2)O(2) that are sufficient to kill other bacterial species is unknown. In the present study, pyruvate oxidase (SpxB), the enzyme responsible for endogenous H(2)O(2) production, was required for survival during exposure to high levels (20 mM) of exogenously added H(2)O(2). Pretreatment with H(2)O(2) did not increase H(2)O(2) resistance in the mutant, suggesting that SpxB activity itself is required, rather than an H(2)O(2)-inducible pathway. SpxB mutants synthesized 85% less acetyl-phosphate, a potential source of ATP. During H(2)O(2) exposure, ATP levels decreased more rapidly in spxB mutants than in wild-type cells, suggesting that the increased killing of spxB mutants was due to more rapid ATP depletion. Together, these data support the hypothesis that S. pneumoniae SpxB contributes to an H(2)O(2)-resistant energy source that maintains viability during oxidative stress. Thus, SpxB is required for resistance to the toxic by-product of its own activity. Although H(2)O(2)-dependent hydroxyl radical production and the intracellular concentration of free iron were similar to that of Escherichia coli, killing by H(2)O(2) was unaffected by iron chelators, suggesting that S. pneumoniae has a novel mechanism to avoid the toxic effects of the Fenton reaction.
authors: Departments of Microbiology, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania 19104, USA.
authors: Christopher D
authors: CD
authors: Pericone
authors: Sunny
authors: S
authors: Park
authors: James A
authors: JA
authors: Imlay
authors: Jeffrey N
authors: JN
authors: Weiser
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism; D024881:Drug Resistance, Bacterial / Q000235:genetics; D006861:Hydrogen Peroxide / Q000032:analysis / Q000494:pharmacology*; D017665:Hydroxyl Radical / Q000378:metabolism; D007501:Iron / Q000032:analysis / Q000378:metabolism*; D009154:Mutation; D010755:Organophosphates / Q000378:metabolism; D018384:Oxidative Stress; D011771:Pyruvate Oxidase / Q000235:genetics / Q000378:metabolism*; D013296:Streptococcus pneumoniae / Q000187:drug effects / Q000201:enzymology* / Q000235:genetics
chemical_list: D010755:Organophosphates; D017665:Hydroxyl Radical; C011632:acetyl phosphate; D000255:Adenosine Triphosphate; D006861:Hydrogen Peroxide; D007501:Iron; D011771:Pyruvate Oxidase