Escherichia coli periplasmic thiol peroxidase acts as lipid hydroperoxide peroxidase and the principal antioxidative function during anaerobic growth.
To clarify the enzymatic property of Escherichia coli periplasmic thiol peroxidase (p20), the specific peroxidase activity toward peroxides was compared with other bacterial thiol peroxidases. p20 has the most substrate preference and peroxidase activity toward organic hydroperoxide. Furthermore, p20 exerted the most potent lipid peroxidase activity. Despite that the mutation of p20 caused the highest susceptibility toward organic hydroperoxide and heat stress, the cellular level of p20 did not respond to the exposure of oxidative stress. Expression level of p20 during anaerobic growth was sustained at the approximately 50% level compared with that of the aerobic growth. Viability of aerobic p20Delta without glucose was reduced to the approximately 65% level of isogenic strains, whereas viability of aerobic p20Delta with 0.5% glucose supplement was sustained. The deletion of p20 resulted in a gradual loss of the cell viability during anaerobic growth. At the stationary phase, the viability of p20Delta was down to approximately 10% level of parent strains. An analysis of the protein carbonyl contents of p20Delta as a marker for cellular oxidation indicates that severe reduction of viability of anaerobic p20Delta was caused by cumulative oxidative stress. P20Delta showed hypersensitivity toward membrane-soluble organic hydroperoxides. An analysis of protein carbonyl and lipid hydroperoxide contents in the membrane of the stress-imposed p20Delta demonstrates that the severe reduction of viability was caused by cumulative oxidative stress on the membrane. Taken together, present data uncover in vivo function for p20 as a lipid hydroperoxide peroxidase and demonstrate that, as the result, p20 acts as the principal antioxidant in the anaerobic habitats.
Cha
Mee-Kyung
MK
Department of Biochemistry, Paichai University, Taejon 302-735, Republic of Korea.
Kim
Won-Cheol
WC
Lim
Chang-Jin
CJ
Kim
Kanghwa
K
Kim
Il-Han
IH
D000693:Anaerobiosis / Q000502:physiology; D000975:Antioxidants / Q000378:metabolism; D004926:Escherichia coli / Q000201:enzymology* / Q000254:growth & development; D029968:Escherichia coli Proteins / Q000502:physiology*; D008054:Lipid Peroxides / Q000378:metabolism; D010084:Oxidation-Reduction; D018384:Oxidative Stress / Q000502:physiology; D033903:Periplasmic Proteins / Q000502:physiology*; D010544:Peroxidases / Q000502:physiology*
D000975:Antioxidants; D029968:Escherichia coli Proteins; D008054:Lipid Peroxides; D033903:Periplasmic Proteins; D010544:Peroxidases; C097049:Tpx protein, E coli