Thioredoxin system in obligate anaerobe Desulfovibrio desulfuricans: Identification and characterization of a novel thioredoxin 2.
Metal corroding sulfate reducing bacteria have been poorly characterized at molecular level due to difficulties pertaining to isolation and handling of anaerobes. We report here for the first time the presence and characterization of thioredoxin 2 in an obligate anaerobic dissimilatory sulfate reducing bacterium Desulfovibrio desulfuricans. In silico analysis of the D. desulfuricans genome revealed the presence of thioredoxin 1 (dstrx1), thioredoxin 2 (dstrx2) and thioredoxin reductase (dstrxR) genes. These genes were found to be actively expressed by the bacteria under the anaerobic growth conditions. We have overexpressed the anaerobic thioredoxin genes in E. coli to produce functionally active recombinant proteins. Recombinant DsTrxR recognized both DsTrx1 and DsTrx2 as its substrate. Mutation studies revealed that the activity of DsTrx2 can be completely abolished with a single amino acid mutation (C69A) in the signature motif 'WCGPC'. Furthermore, the N-terminal domain of DsTrx2 containing two extra CXXC motifs was found to have a negative regulation on its biochemical activity. In conclusion, we have shown the presence of thioredoxin 2 for the first time in an obligate anaerobe which in this anaerobe may be required for its survival under either oxidative stress conditions or metal ion hemostasis.
Department of Biotechnology, All India Institute of Medical Sciences, Ansari Nagar, New Delhi-110029, India.
Ritu
R
Sarin
Yagya D
YD
Sharma
D020816:Amino Acid Motifs / Q000235:genetics; D000693:Anaerobiosis / Q000235:genetics; D001421:Bacteria, Anaerobic / Q000737:chemistry / Q000235:genetics* / Q000254:growth & development; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics*; D045285:Desulfovibrio desulfuricans / Q000737:chemistry / Q000235:genetics* / Q000254:growth & development; D008565:Membrane Proteins / Q000737:chemistry / Q000235:genetics*; D010084:Oxidation-Reduction; D018384:Oxidative Stress / Q000235:genetics*; D017434:Protein Structure, Tertiary / Q000235:genetics; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics; D017386:Sequence Homology, Amino Acid; D013879:Thioredoxins / Q000737:chemistry / Q000235:genetics*
D001426:Bacterial Proteins; D008565:Membrane Proteins; D011994:Recombinant Proteins; D013879:Thioredoxins