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    Purification and characterization of an alkaline pectin lyase produced by a newly isolated brevibacillus borstelensis (p35) and its applications in fruit juice and oil extraction
    (Springer Verlag, 2014) Demir N.; Nadaroglu H.; Demir Y.; Isik C.; Taskin E.; Adiguzel A.; Gulluce M.
    An alkaline pectin lyase (Pnl) (EC 4.2.2.10) secreted by Brevibacillus borstelensis P35 (genBank number: FJ417406) was purifed using ammonium sulfate fractionation, anion exchange chromatography on DEaE-cellulose and gel fltration chromatography on Sephadex g-150. The pH and temperature optima of the enzyme were found to be 8.0 and 60 °C. The enzyme does not loose activity up to 60 °C if exposed for 1 h. The values of Kmand Vmax of the enzyme were 0.625 mg/ml and 126.32 s-1, respectively. The molecular weight was found to be 36 ± 01 kDa. The presence of 10 mM concentration of Ca2+, Cu2+, Mn2+, Mg2+, Zn2+, Hg2+, Fe2+ and EDTa, L-cystein, ascorbic acid signifcantly enhanced the Pnl of the purifed enzyme. In the course of the laboratory trials, it was demonstrated that Pnl from B. borstelensis (P35) could be successfully applied to the production and clarif-cation of fruit juice and oil extraction. © Springer-Verlag Berlin Heidelberg 2014
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    Purification of paraoxonase enzyme from the sera of patients with behcet’s disease and analyzing the effects of the drugs containing imuran (azathioprine), prednisolone (methylprednisolone) and colchium (colchicine)
    (Bentham Science Publishers B.V., 2014) Demir N.; Nadaroglu H.; Ozkan A.; Tasgin E.; Isik C.; Demir Y.
    In this study, serum samples from 50 patients with the diagnosis of Behcet’s disease and 20 healthy volunteers were analyzed. The study consists of three parts. In the first part, paraoxonase (PON) activities were determined in the serum samples of 50 patients with Behcet’s disease and 20 healthy people. In the second part, equal volumes of serum samples from 50 patients were pooled and PON enzymes were purified by using Sepharose-4B-L-tyrosine tyrosine-1-naphtylamine affinity column. Optimum temperature, optimum pH, Vmax and Km values of the pure enzymes were determined. The same purification procedure was also performed in the serum samples of 20 healthy people. Electrophoretic mobility was observed (via SDS-PAGE) in the PON enzymes that were purified from the serum samples of patients with Behcet’s disease and healthy people. In the third part, in vitro effects of drugs containing azathioprine, methylprednisolone and colchicine that have already been used for the treatment of Behcet’s disease were tested on the PON enzymes of the patients with Behcet’s disease and control group. IC50 values and Ki constant values were measured and inhibition types were determined for the drugs containing azathioprine, methylprednisolone and colchicine that have already been used for the treatment of the Behcet’s disease and demonstrate in vitro inhibition effects. ©2014 Bentham Science Publishers.