Putative down-stream signaling molecule of GTPase in Porphyromonas gingivalis

Porphyromonas gingivalis is a strict anaerobic bacterium mainly responsible for periodontal disease in oral cavity. Putative GTPase gene (pgp) of this bacterium was cloned and its recombinant protein (rPGP) was produced in Escherichia coli. Based on the amino acid sequence of SGP that is a GTP-binding protein of Streptococcus mutans, putative GTPase amino acid sequence was deduced in the data base of genome sequences of Porphyromonas gingivalis. A 900-bp PCR fragment was amplified with P. gingivalis genomic DNA as a template and cloned into E. coli JM109. Then pgp was transferred into pQE-30 expression vector to make pQE-PGP for production of rPGP. This protein was produced and purified by Ni-NTA affinity column chromatography. Anti-PGP antibody was also produced in Sprague Dawley rats. Using Westernblot analysis with this antibody, it was confirmed that the rPGP produced in E. coli was identical to that of donor strain. Furthermore, by Southernblot analysis it was revealed that the pgp was originated from P. gingivalis. By immunoprecipitation with anti-PGP antibody and N-terminal amino acid sequence analysis it was found that PGP was able to bind to acetate kinase, which was reported to be a secondary signaling molecule in anaerobic microorganisms. Therefore, these results imply that P. gingivalis produces putative GTPase and this protein might play a potential role in signaling pathway in oral biofilm formation.     

Degradation of Cochlodinium polykrikoides using photocatalytic reactor with TiO2-coated alumina

Cochlodinium polykrikoides (C. polykrikoides) is one of the most harmful red tide dinoflagellates due to its great economic damage and compromising recreational opportunity and public health. Titanium dioxide (TiO₂) is known to be used as a photocatalyst for the control of the aquatic invasive algae under natural and artificial light. The purpose of this study was to design a highly efficient continuous photocatalytic reactor with TiO₂-coated alumina for the demonstrated efficient degradation of C. polykrikoides. TiO₂ photocatalyst beads were prepared by sol-gel dip-coating method using titanium tetra iso-propoxide on alumina. After 40 min of ultraviolet illumination time, the reduction of C. polykrikoides cell number was more than 80%. The degree of degradation of C. polykrikoides increased in a time-dependent manner in this novel reactor. The degradation begins with photocatalytic action by the oxidative species of TiO₂ on the protective cell structures of C. polykrikoides.