Bilous P T, Weiner J H
J Bacteriol. 1985 Jul;163(1):369-75. doi: 10.1128/jb.163.1.369-375.1985.
Proton translocation coupled to dimethyl sulfoxide (DMSO) reduction was examined in Escherichia coli HB101 grown anaerobically on glycerol and DMSO. Rapid acidification of the medium was observed when an anaerobic suspension of cells, preincubated with glycerol, was pulsed with DMSO, methionine sulfoxide, nitrate, or trimethylamine N-oxide. The DMSO-induced acidification was sensitive to the uncoupler carbonyl cyanide p-trifluoromethoxyphenylhydrazone (60 microM) and was inhibited by the quinone analog 2-n-heptyl-4-hydroxy-quinoline-N-oxide (5.6 microM). Neither sodium azide nor potassium cyanide inhibited the DMSO response. An apparent----H+/2e- ratio of 2.9 was obtained for DMSO reduction with glycerol as the reductant. Formate and H2(g), but not lactate, could serve as alternate electron donors for DMSO reduction. Cells grown anaerobically on glycerol and fumarate displayed a similar response to pulses of DMSO, methionine sulfoxide, nitrate, and trimethylamine N-oxide with either glycerol or H2(g) as the electron donor. However, fumarate pulses did not result in acidification of the suspension medium. Proton translocation coupled to DMSO reduction was also demonstrated in membrane vesicles by fluorescence quenching. The addition of DMSO to hydrogen-saturated everted membrane vesicles resulted in a carbonyl cyanide p-trifluoromethoxyphenyl-hydrazone-sensitive fluorescence quenching of quinacrine dihydrochloride. The data indicate that reduction of DMSO by E. coli is catalyzed by an anaerobic electron transport chain, resulting in the formation of a proton motive force.
在以甘油和二甲基亚砜(DMSO)为厌氧生长底物的大肠杆菌HB101中,研究了与DMSO还原偶联的质子转运。当用甘油预孵育的细胞厌氧悬浮液用DMSO、甲硫氨酸亚砜、硝酸盐或三甲胺N-氧化物进行脉冲处理时,观察到培养基迅速酸化。DMSO诱导的酸化对解偶联剂羰基氰对三氟甲氧基苯腙(60微摩尔)敏感,并被醌类似物2-正庚基-4-羟基喹啉-N-氧化物(5.6微摩尔)抑制。叠氮化钠和氰化钾均不抑制DMSO反应。以甘油作为还原剂还原DMSO时,表观的H⁺/2e⁻ 比率为2.9。甲酸盐和H₂(g)而非乳酸盐可作为DMSO还原的替代电子供体。在以甘油和富马酸盐为厌氧生长底物的细胞中,以甘油或H₂(g)作为电子供体时,对DMSO、甲硫氨酸亚砜、硝酸盐和三甲胺N-氧化物脉冲显示出类似的反应。然而,富马酸盐脉冲并未导致悬浮培养基酸化。通过荧光猝灭在膜囊泡中也证明了与DMSO还原偶联的质子转运。向氢饱和的外翻膜囊泡中添加DMSO会导致对羰基氰对三氟甲氧基苯腙敏感的盐酸喹吖因荧光猝灭。数据表明大肠杆菌对DMSO的还原是由厌氧电子传递链催化的,导致质子动力势的形成。