Saunders N F, Hornberg J J, Reijnders W N, Westerhoff H V, de Vries S, van Spanning R J
Department of Molecular Cell Physiology, Faculty of Biology, BioCentrum Amsterdam, Vrije Universiteit, Amsterdam, The Netherlands.
J Bacteriol. 2000 Sep;182(18):5211-7. doi: 10.1128/JB.182.18.5211-5217.2000.
The nos (nitrous oxide reductase) operon of Paracoccus denitrificans contains a nosX gene homologous to those found in the nos operons of other denitrifiers. NosX is also homologous to NirX, which is so far unique to P. denitrificans. Single mutations of these genes did not result in any apparent phenotype, but a double nosX nirX mutant was unable to reduce nitrous oxide. Promoter-lacZ assays and immunoblotting against nitrous oxide reductase showed that the defect was not due to failure of expression of nosZ, the structural gene for nitrous oxide reductase. Electron paramagnetic resonance spectroscopy showed that nitrous oxide reductase in cells of the double mutant lacked the Cu(A) center. A twin-arginine motif in both NosX and NirX suggests that the NosX proteins are exported to the periplasm via the TAT translocon.
反硝化副球菌的nos(一氧化二氮还原酶)操纵子包含一个nosX基因,该基因与在其他反硝化菌的nos操纵子中发现的基因同源。NosX也与NirX同源,而NirX是迄今为止反硝化副球菌所特有的。这些基因的单突变并未导致任何明显的表型,但nosX nirX双突变体无法还原一氧化二氮。启动子-lacZ分析以及针对一氧化二氮还原酶的免疫印迹表明,该缺陷并非由于一氧化二氮还原酶的结构基因nosZ表达失败所致。电子顺磁共振光谱显示,双突变体细胞中的一氧化二氮还原酶缺乏Cu(A)中心。NosX和NirX中的双精氨酸基序表明,NosX蛋白通过TAT转位酶转运到周质中。