Radiation-inducible radioprotective exopolysaccharides of Bacillus siamensis CV5 from irradiated roots of Cistanche violacea to decrease free radical damage produced by ionizing radiation
Purpose: To assess exopolysaccharides (EPS) of Bacillus siamensis CV5, isolated from irradiated roots of Cistanche violacea, for their induction by ionizing radiation (IR) and their antioxidant and radioprotective power.
Materials and methods: Isolated bacteria from the roots of C. violacea were screened for EPS production. The most EPS-producing bacterium was selected and the response surface methodology (RSM) was applied to elucidate the IR dose effects on EPS production. Gamma irradiation effects on the morphology and functional groups of EPS were studied using microscopy and Fourier transform infra-red (FT-IR). The radioprotective potential of EPS on the survival of B. siamensis CV5 following IR was evaluated using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Free radicals scavenging potentialities (FRSP) of non-irradiated and irradiated EPS were evaluated through 2, 2−-diphenyl-1-picrylhydrazyl (DPPH), 2,2′-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid (ABTS) and ferric reducing ability of plasma (FRAP) assays.
Results: Twenty strains, isolated from irradiated roots of C. violacea, were screened for their EPS production. IR-resistant B. siamensis CV5 was the most EPS-producing strain. Its purified EPS contained rhamnose, fructose, mannose and glucose. RSM indicated that EPS of CV5 (CV5-EPS) are radiation inducible. Micrographs of CV5-EPS suggested an increase in the total area and a decrease in the Feret’s statistical diameter following exposure to IR. FT-IR spectra of these EPS revealed an increase of various functional groups. The MTT survival assay demonstrated a positive correlation between the added quantity of CV5-EPS and the viability of irradiated CV5 (p < .01). DPPH, ABTS and FRAP assays indicated that the antioxidant activities of CV5-EPS increased significantly with the irradiation dose (p < .01).
Conclusions: CV5-EPS were demonstrated as radiation-inducible and radioprotective biomolecules. This radioprotective potential of CV5-EPS could be associated with their antioxidant activities. In the future, irradiated EPS could be tested as a gel in cancer radiotherapy for minimizing the damage caused by rays to surrounding healthy tissues.