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锰过氧化氢酶(Alr0998)可保护光合固氮蓝藻鱼腥藻 PCC7120 免受氧化应激。

Mn-catalase (Alr0998) protects the photosynthetic, nitrogen-fixing cyanobacterium Anabaena PCC7120 from oxidative stress.

机构信息

Molecular Biology Division, Bhabha Atomic Research Centre, Mumbai- 400 085, India.

出版信息

Environ Microbiol. 2012 Nov;14(11):2891-900. doi: 10.1111/j.1462-2920.2012.02847.x. Epub 2012 Aug 17.

Abstract

Role of the non-haem, manganese catalase (Mn-catalase) in oxidative stress tolerance is unknown in cyanobacteria. The ORF alr0998 from the Anabaena PCC7120, which encodes a putative Mn-catalase, was constitutively overexpressed in Anabaena PCC7120 to generate a recombinant strain, AnKat(+). The Alr0998 protein could be immunodetected in AnKat(+) cells and zymographic analysis showed a distinct thermostable catalase activity in the cytosol of AnKat(+) cells but not in the wild-type Anabaena PCC7120. The observed catalase activity was insensitive to inhibition by azide indicating that Alr0998 protein is indeed a Mn-catalase. In response to oxidative stress, the AnKat(+) showed reduced levels of intracellular ROS which was also corroborated by decreased production of an oxidative stress-inducible 2-Cys-Prx protein. Treatment of wild-type Anabaena PCC7120 with H(2)O(2) caused (i) RNA degradation in vivo, (ii) severe reduction of photosynthetic pigments and CO(2) fixation, (iii) fragmentation and lysis of filaments and (iv) loss of viability. In contrast, the AnKat(+) strain was protected from all the aforesaid deleterious effect under oxidative stress. This is the first report on protection of an organism from oxidative stress by overexpression of a Mn-catalase.

摘要

在蓝藻中,非血红素、锰过氧化物酶(Mn-catalase)在氧化应激耐受中的作用尚不清楚。来自鱼腥藻 PCC7120 的 ORF alr0998 编码一个假定的 Mn-catalase,该基因在鱼腥藻 PCC7120 中被组成型过表达,以产生重组菌株 AnKat(+)。在 AnKat(+)细胞中可以免疫检测到 Alr0998 蛋白,并且同工酶分析显示 AnKat(+)细胞胞质中存在明显的耐热过氧化物酶活性,但在野生型鱼腥藻 PCC7120 中没有。观察到的过氧化物酶活性对叠氮化物抑制不敏感,表明 Alr0998 蛋白确实是 Mn-catalase。在氧化应激下,AnKat(+)细胞中细胞内 ROS 水平降低,这也得到了氧化应激诱导的 2-Cys-Prx 蛋白产量减少的证实。用 H(2)O(2)处理野生型鱼腥藻 PCC7120 会导致(i)体内 RNA 降解,(ii)光合色素和 CO(2)固定严重减少,(iii)丝状体断裂和裂解,以及(iv)丧失活力。相比之下,AnKat(+)菌株在氧化应激下能免受所有上述有害影响。这是过表达 Mn-catalase 保护生物体免受氧化应激的首例报道。

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