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Identification and analysis of the Shewanella oneidensis major oxygen-independent coproporphyrinogen III oxidase gene

机译:沙瓦氏假单胞菌主要氧非依赖性原卟啉原Ⅲ氧化酶基因的鉴定与分析

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摘要

Shewanella oneidenesis MR-1 is a facultative anaerobe that can use a large number of electron acceptors including metal oxides. During anaerobic respiration, S. oneidensis MR-1 synthesizes a large number of c cytochromes that give the organism its characteristic orange color. Using a modified mariner transposon, a number of S. oneidensis mutants deficient in anaerobic respiration were generated. One mutant, BG163, exhibited reduced pigmentation and was deficient in c cytochromes normally synthesized under anaerobic condition. The deficiencies in BG163 were due to insertional inactivation of hemN1, which exhibits a high degree of similarity to genes encoding anaerobic coproporphyrinogen III oxidases that are involved in heme biosynthesis. The ability of BG163 to synthesize c cytochromes under anaerobic conditions, and to grow anaerobically with different electron acceptors was restored by the introduction of hemN1 on a plasmid. Complementation of the mutant was also achieved by the addition of hemin to the growth medium. The genome sequence of S. oneidensis contains three putative anaerobic coproporphyrinogen III oxidase genes. The protein encoded by hemN1 appears to be the major enzyme that is involved in anaerobic heme synthesis of S. oneidensis. The other two putative anaerobic coproporphyrinogen III oxidase genes may play a minor role in this process.
机译:希瓦氏菌一氧化氮MR-1是一种兼性厌氧菌,可以使用包括金属氧化物在内的大量电子受体。在无氧呼吸过程中,沙门氏菌MR-1合成了大量的c细胞色素,赋予有机体特征性的橙色。使用改良的水手转座子,产生了许多缺乏厌氧呼吸的沙门氏菌突变体。一种突变体BG163表现出减少的色素沉着,并且在通常在厌氧条件下合成的c细胞色素缺乏。 BG163的缺陷归因于hemN1的插入失活,hemN1与编码参与血红素生物合成的厌氧原卟啉原III氧化酶的基因表现出高度相似性。通过在质粒上引入hemN1,恢复了BG163在厌氧条件下合成c细胞色素以及与不同电子受体厌氧生长的能力。还通过向生长培养基中添加血红素来实现突变体的互补。拟南芥的基因组序列包含三个推定的厌氧原卟啉原Ⅲ氧化酶基因。 hemN1编码的蛋白质似乎是参与拟南芥厌氧血红素合成的主要酶。其他两个推定的厌氧性原卟啉原III氧化酶基因可能在此过程中起较小作用。

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