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两类产NO的亚硝酸盐还原酶的分布,结构与序列比较及宏基因组分析
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山东省自然科学基金青年基金(ZR2012CQ011);曲阜师范大学实验教改项目(sj201411)


Distribution, structure and sequence alignment, and metagenomics analysis of two nitrite reductases with NO forming
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    摘要:

    [目的]为了体现亚硝酸盐还原酶在环境中氮生物循环的重要性,研究了它们的分布情况。[方法]利用现有亚硝酸盐还原酶序列在已经测序的基因组数据库中进行查找,研究该酶的分布情况,通过多序列比对比较了它们的序列相似性,通过构建系统发育树比较其进化关系,并利用宏基因组学的方法研究了它们在海洋宏基因组中的分布。[结果]分析结果显示,两类亚硝酸盐还原酶在已测序的细菌和古生菌基因组中分别有397和812个,分别占总量的8%和15.7%,几乎所有的古生菌都含有Ⅱ类酶;它们自身的序列相似性很高,在Ⅰ类酶和Ⅱ类酶中底物结合位点以及Ⅱ类酶的铜离子结合位点保守性都很高,显示该酶序列保守性与其环境功能相适应的特点;进化分析显示它们可能具有共同的进化来源;在海洋宏基因组中,平均每100000读数中分别有6个Ⅰ类和35个Ⅱ类,且2类酶都在热带南太平洋区域有最大分布。[结论]NIR在氮的生物循环及环境修复中可能起到重要作用。

    Abstract:

    [Objective] To reflect the importance of nitrite reductase (NIR) in the environment, we studied its distribution. [Methods] The sequences of NIR were searched in the sequenced genome database at NCBI based on previous reported NIR sequences. The sequence similarity was done by multiple sequence alignment, and phylogenetic relationship was evaluated via constructing the phylogenetic tree. Furthermore, their distribution in the marine metagenome was studied by metagenomics. [Results] The homologues of these two enzymes were 397 and 812 strains in sequenced genome, and the proportion was 8 and 15.7 percent, respectively. Almost all of archaea containing type II NIR. They have high identity by multiple sequence alignment analysis. The cofactor, the substrate and the cooper binding sites in type II were high conserved, suggesting that these enzymes had the specific function in denitrification. Phylogenetic analysis showed the two enzymes may have the common ancestor. In marine metagenome analysis, type I and II have 6 and 35 reads per 100000 reads, respectively, the two types of NIRs have the biggest proportion at the tropical south pacific area. [Conclusion] Collectively, we suggested NIR, especially type II, play a key role in bioremediation of nitrogen contamination.

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辛玉峰,赵天颖,曲晓华. 两类产NO的亚硝酸盐还原酶的分布,结构与序列比较及宏基因组分析. 微生物学报, 2017, 57(4): 597-608

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历史
  • 收稿日期:2016-09-01
  • 最后修改日期:2016-11-03
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  • 在线发布日期: 2017-03-31
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