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首页> 外文期刊>Applied Microbiology >Elimination of Manganese(II,III) Oxidation in Pseudomonas putida GB-1 by a Double Knockout of Two Putative Multicopper Oxidase Genes
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Elimination of Manganese(II,III) Oxidation in Pseudomonas putida GB-1 by a Double Knockout of Two Putative Multicopper Oxidase Genes

机译：通过双重敲除两个推定的多铜氧化酶基因消除恶臭假单胞菌GB-1中锰（II，III）的氧化

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Bacterial manganese(II) oxidation impacts the redox cycling of Mn, other elements, and compounds in the environment; therefore, it is important to understand the mechanisms of and enzymes responsible for Mn(II) oxidation. In several Mn(II)-oxidizing organisms, the identified Mn(II) oxidase belongs to either the multicopper oxidase (MCO) or the heme peroxidase family of proteins. However, the identity of the oxidase in Pseudomonas putida GB-1 has long remained unknown. To identify the P. putida GB-1 oxidase, we searched its genome and found several homologues of known or suspected Mn(II) oxidase-encoding genes ( mnxG , mofA , moxA , and mopA ). To narrow this list, we assumed that the Mn(II) oxidase gene would be conserved among Mn(II)-oxidizing pseudomonads but not in nonoxidizers and performed a genome comparison to 11 Pseudomonas species. We further assumed that the oxidase gene would be regulated by MnxR, a transcription factor required for Mn(II) oxidation. Two loci met all these criteria: PputGB1_2447, which encodes an MCO homologous to MnxG, and PputGB1_2665, which encodes an MCO with very low homology to MofA. In-frame deletions of each locus resulted in strains that retained some ability to oxidize Mn(II) or Mn(III); loss of oxidation was attained only upon deletion of both genes. These results suggest that PputGB1_2447 and PputGB1_2665 encode two MCOs that are independently capable of oxidizing both Mn(II) and Mn(III). The purpose of this redundancy is unclear; however, differences in oxidation phenotype for the single mutants suggest specialization in function for the two enzymes.
机译：细菌锰（II）的氧化影响环境中Mn，其他元素和化合物的氧化还原循环;因此，了解Mn（II）氧化的机理和酶很重要。在几种Mn（II）氧化生物中，鉴定出的Mn（II）氧化酶属于多铜氧化酶（MCO）或血红素过氧化物酶家族的蛋白质。然而，恶臭假单胞菌GB-1中氧化酶的身份长期以来仍是未知的。为了鉴定恶臭假单胞菌GB-1氧化酶，我们搜索了它的基因组，发现了已知或怀疑的Mn（II）氧化酶编码基因（mnxG，mofA，moxA和mopA）的几个同源物。为了缩小清单范围，我们假设Mn（II）氧化酶基因在Mn（II）氧化假单胞菌中会保守，但在非氧化剂中不会保守，并与11个假单胞菌种进行了基因组比较。我们进一步假设氧化酶基因将受MnxR（Mn（II）氧化所需的转录因子）调控。有两个基因座符合所有这些条件：PputGB1_2447，其编码与MnxG同源的MCO; PputGB1_2665，其编码与MofA具有非常低的同源性的MCO。每个基因座的框内缺失导致菌株保留了一定的氧化Mn（II）或Mn（III）的能力。仅当两个基因都缺失时才达到氧化损失。这些结果表明，PputGB1_2447和PputGB1_2665编码两个MCO，它们分别能够氧化Mn（II）和Mn（III）。这种冗余的目的尚不清楚。然而，单个突变体在氧化表型上的差异表明这两种酶的功能专门化。

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