[ Objective] The research aimed to get the optimized separation and purification conditions of the hirudin produced from Bacillus subtilis DB403 （pUBH5）. [Method] Through the systemic pretreatment, preliminary chrom...[ Objective] The research aimed to get the optimized separation and purification conditions of the hirudin produced from Bacillus subtilis DB403 （pUBH5）. [Method] Through the systemic pretreatment, preliminary chromatography and fine chromatography. [Result]The optimized separation and purification conditions were that： Supernatant was treated by trichloroacetic acid, then by ultrafiltration desalt and anion exchange chromatography. Strong anion Q F. F. was better than weak anion DEAE F.F. The proper balanced solution was Tris-HCI （ pH 8.0）. The proper conductivity was 6 ms/cm. The maximum applied sample was 240 ATU/ml to matrix of strong anion Q F. F. This optimized procedure was magnified in strong anion exchange HiPrep 16/10Q with the 90% recovery and 70.2% purity. The purification of gel filtration of Sephacryl S-100 to hirudin was not relative to flow rate within certain scope. The application size of sample was 10 ml. The purity checked by HPLC was 95.1%, and the recovery was 93%, and the band of SDS-PAGE was single. [ Conclusion] The research provided the reference of the further industrialization separation and purification of hiruin.展开更多
A new isoenzyme of tobacco peroxidase(TOP) I was purified from tobacco (K326) by using acetone powder, ammonium sulfate precipitation and column chromatography on DEAE-52 cellulose, Sephadex G-75 and DEAE-Sephadex A-5...A new isoenzyme of tobacco peroxidase(TOP) I was purified from tobacco (K326) by using acetone powder, ammonium sulfate precipitation and column chromatography on DEAE-52 cellulose, Sephadex G-75 and DEAE-Sephadex A-50. It is an iron-protein containing haemachrome, whose molecular weight is 21888.5 and the isoelectric point is 3.5. The optimum pH value and temperature of this enzyme is 6.0 and 45℃respectively. The enzyme is stable in the pH range from 3.0 to 10.0 and has a favorable thermostability.展开更多
In this study, the separation and purification of Cordyceps sinensis polysaccharides by AB-8 and D-280 macro porous ad- sorption resin were researched, the adsorption characters when AB-8 macro porous adsorption resin...In this study, the separation and purification of Cordyceps sinensis polysaccharides by AB-8 and D-280 macro porous ad- sorption resin were researched, the adsorption characters when AB-8 macro porous adsorption resin processed Cordyceps sinensis leachate were determined, and the eluent were bleaching treated by D-280 macro porous adsorption resin. The results showed that the concentration of 2.0mg/mL, pH = 6 at a flow rate of 0.8mL/min. The yielding amount of polysaccharides is 246.4mg, and after decolouring treatment, the final yield is 207.1 mg.展开更多
基金Supported by 863 Program of China(2006AA03Z0453)NaturalScience Research Program of Higher Education of Jiangsu Province(09KJB230001)+1 种基金973 Program of China(2009CB724700)AndSchool Foundation of Jiangsu University(08JDG009)~~
文摘[ Objective] The research aimed to get the optimized separation and purification conditions of the hirudin produced from Bacillus subtilis DB403 （pUBH5）. [Method] Through the systemic pretreatment, preliminary chromatography and fine chromatography. [Result]The optimized separation and purification conditions were that： Supernatant was treated by trichloroacetic acid, then by ultrafiltration desalt and anion exchange chromatography. Strong anion Q F. F. was better than weak anion DEAE F.F. The proper balanced solution was Tris-HCI （ pH 8.0）. The proper conductivity was 6 ms/cm. The maximum applied sample was 240 ATU/ml to matrix of strong anion Q F. F. This optimized procedure was magnified in strong anion exchange HiPrep 16/10Q with the 90% recovery and 70.2% purity. The purification of gel filtration of Sephacryl S-100 to hirudin was not relative to flow rate within certain scope. The application size of sample was 10 ml. The purity checked by HPLC was 95.1%, and the recovery was 93%, and the band of SDS-PAGE was single. [ Conclusion] The research provided the reference of the further industrialization separation and purification of hiruin.
文摘A new isoenzyme of tobacco peroxidase(TOP) I was purified from tobacco (K326) by using acetone powder, ammonium sulfate precipitation and column chromatography on DEAE-52 cellulose, Sephadex G-75 and DEAE-Sephadex A-50. It is an iron-protein containing haemachrome, whose molecular weight is 21888.5 and the isoelectric point is 3.5. The optimum pH value and temperature of this enzyme is 6.0 and 45℃respectively. The enzyme is stable in the pH range from 3.0 to 10.0 and has a favorable thermostability.
文摘In this study, the separation and purification of Cordyceps sinensis polysaccharides by AB-8 and D-280 macro porous ad- sorption resin were researched, the adsorption characters when AB-8 macro porous adsorption resin processed Cordyceps sinensis leachate were determined, and the eluent were bleaching treated by D-280 macro porous adsorption resin. The results showed that the concentration of 2.0mg/mL, pH = 6 at a flow rate of 0.8mL/min. The yielding amount of polysaccharides is 246.4mg, and after decolouring treatment, the final yield is 207.1 mg.
