CLARiTY 17 UV/Vis用于“大肠杆菌的NADH依赖亚硝酸还原酶Nir在发酵氨化中的作用”

CLARiTY 17 UV/Vis用于“大肠杆菌的NADH依赖亚硝酸还原酶Nir在发酵氨化中的作用”

The role of the NADH-dependent nitrite reductase, Nir, from Escherichia coli in fermentative ammonification

了解每种参与还原酶的特定作用对于描述决定细胞对其环境的反应的生化平衡是必要的,硝酸盐和亚硝酸盐还原对于氮同化和无氧代谢至关重要。大肠杆菌中的可溶性细胞质siroheme NADH-亚硝酸还原酶(Nir)是硝酸盐/亚硝酸盐同化所必需的,但也有报道要么“解毒”亚硝酸盐,要么进行发酵氨化以支持厌氧分解代谢。从理论上讲,亚硝酸盐解毒对于硝酸盐的厌氧生长很重要,在此期间过量的亚硝酸盐会被还原成铵。 Nir的发酵氨化对于在亚硝酸盐存在下的厌氧生长期间非呼吸ATP产生的最大化是重要的。本文报道的实验旨在通过在限定培养基中厌氧条件下培养大肠杆菌以及缺乏Nir或呼吸亚硝酸还原酶(Nrf)的突变菌株,同时监测氮利用和发酵代谢物,来测试Nir在发酵氨化中的潜在作用。为了关注Nir在发酵氨化中的作用,在大多数实验中使用pH控制来消除由于硝酸形成引起的亚硝酸盐毒性。我们的研究结果表明,Nir在发酵生长期间具有显着的益处,反映了发酵氨化而不是解毒。我们得出结论,Nir的发酵氨化允许葡萄糖在能量上有利地发酵成甲酸盐和乙酸盐。根据Nir在其他细菌和植物中的作用,讨论了这些结果和结论。

Nitrate and nitrite reduction are of paramount importance for nitrogen assimilation and anaerobic metabolism, and understanding the specific roles of each participating reductase is necessary to describe the biochemical balance that dictates cellular responses to their environments. The soluble, cytoplasmic siroheme NADH-nitrite reductase (Nir) in Escherichia coli is necessary for nitrate/nitrite assimilation but has also been reported to either “detoxify” nitrite, or to carry out fermentative ammonification in support of anaerobic catabolism. Theoretically, nitrite detoxification would be important for anaerobic growth on nitrate, during which excess nitrite would be reduced to ammonium. Fermentative ammonification by Nir would be important for maximization of non-respiratory ATP production during anaerobic growth in the presence of nitrite. Experiments reported here were designed to test the potential role of Nir in fermentative ammonification directly by growing E. coli along with mutant strains lacking Nir or the respiratory nitrite reductase (Nrf) under anaerobic conditions in defined media while monitoring nitrogen utilization and fermentation metabolites. To focus on the role of Nir in fermentative ammonification, pH control was used in most experiments to eliminate nitrite toxicity due to nitric acid formation. Our results demonstrate that Nir confers a significant benefit during fermentative growth that reflects fermentative ammonification rather than detoxification. We conclude that fermentative ammonification by Nir allows for the energetically favorable fermentation of glucose to formate and acetate. These results and conclusions are discussed in light of the roles of Nir in other bacteria and in plants.


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