In screening for marine bacterial polysaccharide lytic enzymes, the most potent isolate was identified as Bacillus cereus NRC-20. This strain showed high alginase, dextrinase, pectinase and carboxymethylecellulase productivity when production medium was adjusted at pH 6.0 and contains (g/l): 3.0 sodium alginate, 3.0 malt extract, 30.0 jatropha cake and inoculated with 8% (v/v) of a 24 h old cell suspension and incubated at 30°C for 48 h on a rotary shaker at 200 rpm. These enzymes mixture were precipitated by 60% acetone and purified by using sephadex G-100, whereas SDS-PAGE suggested a molecular weight of approximately 17, 20, 89 and 279 KDa, respectively. The optimum partially purified enzymes activity when enzyme protein concentration was 27.4 mg/ml and substrate concentration 5.0 g/l shown after 90 min of incubation at 30°C and pH 5 and stable at 60°C for 60 min. The enzymes activities were enhanced in the presence of 0.02 M Na + and K + ions and completely inactivated by Mg +2 , Cu +2 , Co +2 and Pb +2 The enzymes mixture affected the matrix integrity of different microbial biofilms artificially grown on stainless steel sheets of Bacillus subtilis NRRL B-4219, Staphylococcus aureus ATCC 29213, Pseudomonas aeruginosa ATCC 27953 and Escherichia coli ATCC 25922.
A novel Cr(VI)-resistant haloalkaliphilic bacterial strain NRC-R, identified as Salipaludibacillus agaradhaerens, was isolated from hypersaline soda lakes and characterized for its Cr(VI) bioreduction efficiency. Strain NRC-R grew well and effectively reduced Cr(VI) under a wide range of sodium chloride, pH, shaking velocity and temperature, showing maximum Cr(VI) reduction at 8% NaCl, pH 10, 150 rpm and 35 °C, respectively. Strain NRC-R was able to grow and reduce Cr(VI) effectively in the presence of different heavy metals and oxyanions (Pb 2+ , Zn 2+ , Co 2+ , Mn 2+ , Ni 2+ , Mo 2+ , HPO 4 − , NO 3 − , SO 4 2− and HCO 3 − ). Furthermore, Fe 3+ and Cu 2+ significantly enhanced the Cr(VI) removal by about 1.5 fold. Strain NRC-R could reduce Cr(VI) using a variety of electron donors, exhibiting a maximum reduction in the presence of NADH and fructose. The bioremoval of Cr(VI) using strain NRC-R was due to direct enzymatic reduction and the chromate reductase activity was mainly detected in the bacterial cell membrane. Under the optimized conditions, strain NRC-R showed a remarkable Cr(VI) bioreduction with highest reduction rate of 240 uM/h. Cr(VI) concentrations of up to 3 mM (888.5 mg/L) and 4 mM (1177 mg/L) were completely reduced within 16 h and 32 h, respectively. TEM and SEM-EDX analyses confirmed the biosorption of chromium species into the cells. To the best of our knowledge, this is the first report about Cr(VI) reduction by S. agaradhaerens. In conclusion, S. agaradhaerens NRC-R was a highly efficient Cr(VI) reducing haloalkaliphilic bacterium that has a significant potential in the bioremediation of Cr(VI)-contaminated environments.
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