Nitrite reductase activity in F420-dependent sulphite reductase (Fsr) from Methanocaldococcus jannaschii.

Christian Heryakusuma, Eric F Johnson, Endang Purwantini, Biswarup Mukhopadhyay
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Abstract

Methanocaldococcus jannaschii (Mj), a hyperthermophilic and evolutionarily deeply rooted methanogenic archaeon from a deep-sea hydrothermal vent, produces F420-dependent sulphite reductase (Fsr) in response to exposure to sulphite. This enzyme allows Mj to detoxify sulphite, a potent inhibitor of methyl coenzyme-M reductase (Mcr), by reducing it to sulphide with reduced coenzyme F420 (F420H2) as an electron donor; Mcr is essential for energy production for a methanogen. Fsr allows Mj to utilize sulphite as a sulphur source. Nitrite is another potent inhibitor of Mcr and is toxic to methanogens. It is reduced by most sulphite reductases. In this study, we report that MjFsr reduced nitrite to ammonia with F420H2 with physiologically relevant K m values (nitrite, 8.9 µM; F420H2, 9.7 µM). The enzyme also reduced hydroxylamine with a K m value of 112.4 µM, indicating that it was an intermediate in the reduction of nitrite to ammonia. These results open the possibility that Mj could use nitrite as a nitrogen source if it is provided at a low concentration of the type that occurs in its habitat.

Abstract Image

Abstract Image

jannaschi甲烷钙球菌f420依赖性亚硫酸盐还原酶(Fsr)的亚硝酸盐还原酶活性。
jannaschii甲烷醛球菌(Methanocaldococcus jannaschii, Mj)是一种来自深海热液喷口的嗜热性深根产甲烷古菌,在接触亚硫酸盐时产生依赖f420的亚硫酸盐还原酶(Fsr)。该酶允许Mj解毒亚硫酸盐,甲基辅酶- m还原酶(Mcr)的有效抑制剂,通过还原辅酶F420 (F420H2)作为电子供体将其还原为硫化物;Mcr对产甲烷菌的能量生产至关重要。Fsr允许Mj利用亚硫酸盐作为硫源。亚硝酸盐是另一种有效的Mcr抑制剂,对产甲烷菌有毒。它被大多数亚硫酸盐还原酶还原。在这项研究中,我们报告了MjFsr用F420H2将亚硝酸盐还原为氨,其K m值与生理相关(亚硝酸盐8.9µm;F420h2, 9.7µm)。该酶还能还原羟胺,K m值为112.4µm,表明它是亚硝酸盐还原为氨的中间体。这些结果打开了一种可能性,即Mj可以利用亚硝酸盐作为氮源,如果它在其栖息地中以低浓度的类型提供。
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