Hashimoto Light Energy Conversion Project, ERATO/JST, Komaba Open Laboratory, The University of Tokyo, Tokyo, Japan.
FEMS Microbiol Lett. 2012 Jan;326(1):91-8. doi: 10.1111/j.1574-6968.2011.02444.x. Epub 2011 Nov 14.
Dissimilatory metal-reducing bacteria (DMRB), such as Shewanella oneidensis MR-1, are of great interest for their importance in the biogeochemical cycling of metals and utility in biotechnological processes, such as bioremediation and microbial fuel cells. To identify genes necessary for metal reduction, this study constructed a random transposon-insertion mutant library of MR-1 and screened it for isolating mutants that were deficient in metal reduction. Examination of approximately 5000 mutants on lactate minimal-medium plates containing MnO(2) resulted in the isolation of one mutant, strain N22-7, that showed a decreased MnO(2)-reduction activity. Determination of a transposon-insertion site in N22-7 followed by deletion and complementation experiments revealed that the disruption of SO3030, a siderophore biosynthesis gene, was responsible for the decreased MnO(2)-reduction activity. In ΔSO3030 cells, iron and cytochrome contents were decreased to approximately 50% of those in the wild-type cells, when they were incubated under MnO(2)-reduction conditions. In addition, the transcription of genes encoding outer-membrane cytochromes necessary for metal reduction was repressed in ΔSO3030 under MnO(2)-reduction conditions, while their transcription was upregulated after supplementation of culture media with ferrous iron. These results suggest that siderophore is important for S. oneidensis MR-1 to respire MnO(2), because iron availability influences the expression of cytochromes necessary for metal reduction.
异化金属还原菌(DMRB),如希瓦氏菌属 oneidensis MR-1,因其在金属的生物地球化学循环中的重要性以及在生物技术过程中的应用而备受关注,例如生物修复和微生物燃料电池。为了确定金属还原所需的基因,本研究构建了 MR-1 的随机转座子插入突变体文库,并对其进行筛选以分离金属还原缺陷的突变体。在含有 MnO2 的乳酸最小培养基平板上对大约 5000 个突变体进行检查,导致分离出一个突变体菌株 N22-7,其 MnO2 还原活性降低。在 N22-7 中确定转座子插入位点,然后进行缺失和互补实验,结果表明,铁载体生物合成基因 SO3030 的破坏是 MnO2 还原活性降低的原因。在 ΔSO3030 细胞中,当在 MnO2 还原条件下孵育时,铁和细胞色素的含量降低到野生型细胞的约 50%。此外,在 MnO2 还原条件下,ΔSO3030 中编码金属还原所需的外膜细胞色素的基因转录受到抑制,而在添加亚铁的培养基后,其转录上调。这些结果表明,铁载体对于 S. oneidensis MR-1 呼吸 MnO2 很重要,因为铁的可用性会影响金属还原所需细胞色素的表达。