Study of biofilm-influenced corrosion on X80 pipeline steel by a nitrate-reducing bacterium, Bacillus cereus, in artificial Beijing soil.

The biofilm of Bacillus cereus on the surface of X80 pipeline steel was investigated from forming to shedding. Based on the observed biofilm morphology and pit analysis, it was found that B. cereus biofilm could stimulate X80 pipeline steel pitting corrosion, which was attributed to the nitrate reduction of bacteria beneath the biofilm. Electrochemical measurements and general corrosion rate results showed that B. cereus biofilm can better accelerate X80 pipeline steel corrosion compared to sterile solutions. Interestingly, the results also showed that thick biofilms had a slight tendency to inhibit the general corrosion process compared with its formation and exfoliation, which was confirmed by scanning Kelvin probe. The corrosion rate of X80 pipeline steel in artificial Beijing soil is closely related to the state of the biofilm, and nitrate reducing bacteria accelerates the occurrence of pits. The corresponding corrosion mechanisms are proposed.

A spectroscopic study of uranyl-cytochrome b5/cytochrome c interactions.

Uranium is harmful to human health due to its radiation damage and the ability of uranyl ion (UO2(2+)) to interact with various proteins and disturb their biological functions. Cytochrome b5 (cyt b5) is a highly negatively charged heme protein and plays a key role in mediating cytochrome c (cyt c) signaling in apoptosis by forming a dynamic cyt b5-cyt c complex. In previous molecular modeling study in combination with UV-Vis studies, we found that UO2(2+) is capable of binding to cyt b5 at surface residues, Glu37 and Glu43. In this study, we further investigated the structural consequences of cyt b5 and cyt c, as well as cyt b5-cyt c complex, upon uranyl binding, by fluorescence spectroscopic and circular dichroism techniques. Moreover, we proposed a uranyl binding site for cyt c at surface residues, Glu66 and Glu69, by performing a molecular modeling study. It was shown that uranyl binds to cyt b5 (KD=10 µM), cyt c (KD=87 µM), and cyt b5-cyt c complex (KD=30 µM) with a different affinity, which slightly alters the protein conformation and disturbs the interaction of cyt b5-cyt c complex. Additionally, we investigated the functional consequences of uranyl binding to the protein surface, which decreases the inherent peroxidase activity of cyt c. The information of uranyl-cyt b5/cyt c interactions gained in this study likely provides a clue for the mechanism of uranyl toxicity.

Observation of heme transfer from cytochrome b5 to DNA aptamer.

Heme transfer is commonly observed from one heme protein to the other such as from cytochrome b(5) (cyt b(5)) to apo-myoglobin. In this study, instead of to another heme protein, we observed the heme transfer from wild-type (WT) cyt b(5), H39C cyt b(5) with heme axial ligand His39 mutated to Cys39, and DME cyt b(5) with heme replaced by protoporphyrin IX dimethyl ester, to a heme DNA aptamer, PS2.M, respectively, with a different rate constant. The heme transfer was further confirmed by the enhancement of peroxidase activity of the cyt b(5)s-PS2.M system due to the formation of catalytic PS2.M-heme complex. This study provides valuable insights into both cyt b(5)-heme and PS2.M-heme interactions and shows that heme transfer from heme protein to heme-aptamer can be used to evaluate the relative stability of heme proteins. In addition, this study sheds light on the maturation of heme proteins in vivo by interacting with DNA/RNA enzymes.