Sürdürülebilir Kalkınma Amaçları
Yayınlar
- MakaleAçık Erişim2014Production of pectin lyase from Geobacillus pallidus p26, purification, characterization and fruit juice application
Çelik, Safinur Yildirim; Demir, Nazan; Demir, Yasar; Adiguzel, Ahmet; Gulluce, Medine
A bacterial strain was isolated from Pasinler hot spring, Erzurum, Turkey. The purified thermophilic isolate was identified as Geobacillus pallidus P26 and used to produce extracellular pectin lyase (EC 4.2.2.10). Pectin lyase enzyme was purified 34 fold by using DEAE-cellulose anion exchange column chromatography and characterized. Molecular weight of the enzyme was determined as 56 kDa by using Sephadex G-100 gel filtration chromatography. Purification of enzyme was verified by SDS-PAGE. The pH- and temperature optima of enzyme were determined (pH 9.0 and 60 degrees C, respectively). Pectin lyase was mostly stable at 50 degrees C: for 24 hours. Its' activity ducruitscd to 50 % for 24 h at 60 degrees C. K-M and V-max were calculated as 24.8 mg/mL and 2.28 mu mol/L min, respectively. Purified pectin lyase was inhibited by Fe3+, Zn2+, Cu2+, Ca2+, Co2+ and Hg2+ but not by Mg2+. The purified pectin lyase enzyme was used for getting fruits juices. It was found that yields of fruits juices increased when they were compared with control.
AtıfWoS9OpenAlex9
İncele - MakaleYalnız künye2014Scopus SJRQ2Purification 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)
Demir, Nazan; Nadaroglu, Hayrunnisa; Ozkan, Asuman; Tasgin, Esen; Isik, Ceyhun; Demir, Yasar
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.
AtıfScopus2OpenAlex3
İncele - MakaleYalnız künye2014Scopus SJRQ4Production of pectin lyase from xanthomonas campestris, purification, characterization and fruit juice application
Celik, Safinur Yildirim; Demir, Yasar; Demir, Nazan; Güllüce, Medine
Extracellular pectin lyase (EC 4.2.2.10) was produced by Xanthomonas campestris in solid state fermentation. Solid state fermentation was carried out using 250 mL Erlenmeyer flasks containing 5 g wheat bran, 1% citrus pectin and 10 mL salt mixture that composed of 0.14% (NH4)2SO4 0.02% MgCl2 and 0.02% K2HPO4. Pectin lyase enzyme was purified 43 fold by using DEAE-cellulose anion exchange column chromatography and characterized. Molecular weight of the enzyme was determined as 77.5 kDa by using Sephadex G-100 gel filtration chromatography. Purification of enzyme was controlled by SDS-PAGE. The optimum pH and temperature of enzyme was determined as pH 9.0 and 50°C, respectively. Pectin lyase was mostly stable 40°C for 24 hours. KM and Vmax were calculated respectively as 0.69 mg/mL and 3.84 |μmol/L∗min. Purified pectin lyase was inhibited by Fe+, Zn2+, Ca2+, Co2+ and Hg2+ except for Cu2+ and Mg2+ The purified pectin lyase enzyme was used for getting fruits juices. It was determined that yields of fruits juices improved when it was compared with control.
- MakaleYalnız künye2014Scopus SJRQ1Purification and characterization of an alkaline pectin lyase produced by a newly isolated brevibacillus borstelensis (p35) and its applications in fruit juice and oil extraction
Demir, Nazan; Nadaroglu, Hayrunnisa; Demir, Yasar; Isik, Ceyhun; Taskin, Esen; Adiguzel, Ahmet; Gulluce, Medine
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.
AtıfScopus36WoS31OpenAlex35İncele - MakaleYalnız künye2012Scopus SJRQ4Expression of cytosolic and noncytosolic carbonic anhydrase enzymes from bovine brain membrane
Demir, Nazan; Demir, Yasar; Coskun, Fatma; Tasgin, Esen
Carbonic anhydrase is an enzyme that takes responsibility in inhalation function but, until today, carbonic anhydrase is not examined if it is present in the bovine brain membrane or not. The enzyme carbonic anhydrase was purified and separately characterized according to the bonding forms in 4 steps such as outer peripheral, cytosolic, inner peripheral and integral. Affinity chromatography was used for purification of the enzyme in all four steps. The affinity column was prepared with sepharose-4B-L-tyrosine-sulphanilamide. Purified carbonic anhydrase was obtained at each step. Enzyme activity was measured by CO2 hydratase activity and esterase activity methods. Optimum pH and optimum temperature were defined for purified enzymes at each step. Morover molecular weight and purity were detected by gel filtration and SDS-PAGE electrophorose. In addition, the enzyme's Km and nmax values were found with the Lineweaver- Burk method. The obtained results are discussed in comparison with other mammalian carbonic anhydrases. Carbonic anhydrase was shown to be exist in bovine brain membrane and this enzyme was optimized.
AtıfScopus1İncele - MakaleYalnız künye2009Scopus SJRQ3Purification and properties of carbonic anhydrase from bone marrow
Tasgin, Esen; Nadaroglu, Hayrunnisa; Demir, Yasar; Demir, Nazan
In this work, the carbonic anhydrase was purified from bovine bone marrow and investigated its kinetic properties. Carbonic anhydrase was purified from bovine bone marrow using affinity chromatography by sepharose 4B-L-tyrosine sulphanilamide. During purification steps, the activity of enzyme was measured using p-nitrophenyl acetate at pH: 7.4. Optimum pH and optimum temperature values for bovine bone marrow carbonic anhydrase were determined and then K m and Vmax values for the same substrate were obtained by means of Linewearver-Burk graphics. The purification degree for bovine bone marrow was calculated. The Vmax, and Km values at optimum pH and at 20 °C for the substrate (p-nitrophenyl acetate) were 120.418 uμmol/L min and 2.409 x 103 mM, respectively. The K1 and I50 values for sulfanilamide, KSCN, NaN3 and acetazolamide were determined in bovine bone marrow carbonic anhydrase.
AtıfScopus7İncele