Caffrey Sean M, Park Hyung Soo, Been Jenny, Gordon Paul, Sensen Christoph W, Voordouw Gerrit
University of Calgary, Department of Biological Sciences, 2500 University Drive NW, Calgary, Alberta T2N 1N4, Canada.
Appl Environ Microbiol. 2008 Apr;74(8):2404-13. doi: 10.1128/AEM.02469-07. Epub 2008 Feb 29.
The genome sequence of the sulfate-reducing bacterium Desulfovibrio vulgaris Hildenborough was reanalyzed to design unique 70-mer oligonucleotide probes against 2,824 probable protein-coding regions. These included three genes not previously annotated, including one that encodes a c-type cytochrome. Using microarrays printed with these 70-mer probes, we analyzed the gene expression profile of wild-type D. vulgaris grown on cathodic hydrogen, generated at an iron electrode surface with an imposed negative potential of -1.1 V (cathodic protection conditions). The gene expression profile of cells grown on cathodic hydrogen was compared to that of cells grown with gaseous hydrogen bubbling through the culture. Relative to the latter, the electrode-grown cells overexpressed two hydrogenases, the hyn-1 genes for [NiFe] hydrogenase 1 and the hyd genes, encoding [Fe] hydrogenase. The hmc genes for the high-molecular-weight cytochrome complex, which allows electron flow from the hydrogenases across the cytoplasmic membrane, were also overexpressed. In contrast, cells grown on gaseous hydrogen overexpressed the hys genes for [NiFeSe] hydrogenase. Cells growing on the electrode also overexpressed genes encoding proteins which promote biofilm formation. Although the gene expression profiles for these two modes of growth were distinct, they were more closely related to each other than to that for cells grown in a lactate- and sulfate-containing medium. Electrochemically measured corrosion rates were lower for iron electrodes covered with hyn-1, hyd, and hmc mutant biofilms than for wild-type biofilms. This confirms the importance, suggested by the gene expression studies, of the corresponding gene products in D. vulgaris-mediated iron corrosion.
对硫酸盐还原菌希登伯勒脱硫弧菌(Desulfovibrio vulgaris Hildenborough)的基因组序列进行了重新分析,以针对2824个可能的蛋白质编码区域设计独特的70聚体寡核苷酸探针。其中包括三个先前未注释的基因,其中一个编码c型细胞色素。使用印有这些70聚体探针的微阵列,我们分析了在铁电极表面施加 -1.1 V负电位(阴极保护条件)产生的阴极氢上生长的野生型希登伯勒脱硫弧菌的基因表达谱。将在阴极氢上生长的细胞的基因表达谱与通过向培养物中鼓入气态氢生长的细胞的基因表达谱进行了比较。相对于后者,电极生长的细胞过表达了两种氢化酶,即[NiFe]氢化酶1的hyn-1基因和编码[Fe]氢化酶的hyd基因。允许电子从氢化酶穿过细胞质膜流动的高分子量细胞色素复合物的hmc基因也过表达。相比之下,在气态氢上生长的细胞过表达了[NiFeSe]氢化酶的hys基因。在电极上生长的细胞还过表达了编码促进生物膜形成的蛋白质的基因。尽管这两种生长模式的基因表达谱不同,但它们彼此之间的关系比与在含乳酸和硫酸盐的培养基中生长的细胞的基因表达谱更密切。覆盖有hyn-1、hyd和hmc突变生物膜的铁电极的电化学测量腐蚀速率低于野生型生物膜。这证实了基因表达研究表明的相应基因产物在希登伯勒脱硫弧菌介导的铁腐蚀中的重要性。