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Microbial polysaccharides, due to their unique physiochemical properties, have found application in the food industry, cosmetics, pharmacy and medicine. In the environment, microbes can use polysaccharides to alleviate the adverse effects of heavy metals in their close proximity. This adaptive property shows interesting potential for bioremediation. Herein, the effects of the exopolysaccharides (EPS) levan, produced by the bacterium Bacillus licheniformis NS032 and pullulan, produced by the fungus Aureobasidium pullulans CH-1 in the presence of copper (Cu2+) have been investigated for the first time on antioxidant enzyme activity, respiration and Cu2+ bioaccumulation of Daphnia magna as well as the bioluminescence of Vibrio fischeri. Both EPS decreased toxicity of Cu2+ in the acute test with D. magna. The activity of catalase (CAT) was significantly diminished after acute exposure to Cu2+ in comparison to treatments with Cu2+ and EPS, while in the prolonged acute exposure the CAT activity did not show statistically significant (P ≤ 0.05) differences between treatments with and without the EPS. According to ICP-MS results, during prolonged acute exposure of neonates, the bioaccumulation of Cu2+ in treatments without the EPS was 52.03 μg/g of biomass (wet), while in treatments with EPS, the bioaccumulation was lower by one order of magnitude. The respiration of neonates during acute exposure to Cu2+ with or without the EPS was monitored using the MicroOxymax respirometer, and the results show the EPS can positively effect the respiration. In the case of bacterial bioluminescence, the toxicity of Cu2+ decreased in treatments with EPS (30 min EC10) from 3.54 mg/L to 140.61 mg/L (levan) and 45.00 mg/L (pullulan). This study demonstrates protective effect of EPS against Cu2+ toxicity on D. magna and V. fischeri, and opens the door for further investigation of potential application of levan and pullulan in bioremediation of heavy metals and mitigation of their adverse effects in the aquatic environment.
copper, levan, Vibrio fischeri, pullulan, Daphnia magna, respirometer
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The structure of pullulan, the extracellular α-D-glucan elaborated by the yeast-like fungus Aureobasidium pullulans, may be described as a linear a-D-glucan consisting of maltotriosyl repeat units connected terminally by (1→6)-α-D-glucosidic bonds. Occasionally some of maltotriosyl residues are replaced by higher oligosaccharide units, most frequently with maltotetraosyl residues. Using the susceptibility of pullulan CH-1 (obtained from strain CH-1 of Aureobasidium pullulans) to hydrolysis catalysed by porcine α-amylase, the polysaccharide was cleaved and the fragments obtained fractionated by gel-permeation chromatography. The heterogenous size of the fragments indicates that there is no apparent regular distribution of tetrasaccharide units in the pullulan chain. Enzymatic digestion of pullulan CH-1 using pullulanase, followed by gel-permeation chromatography of the resulting digest confirmed these results as did preparative paper chromatography and CI mass spectrometry of the separated components, i.e., that maltotetraosyl units (about 7 %) are building units of pullulan CH-1.
polysaccharide, pullulan, Aureobasidium pullulans, amylolysis, pullulanolysis
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