新型离子液体亲和萃取鸡蛋清中的溶菌酶

建立并优化利用辛巴蓝(Cibacron Blue F-3GA,CB)修饰C₄MIM]Cl的新型离子液体C₄MIM]₃CB特异性萃取分离鸡蛋清中溶菌酶的方法,并采用AutoDock Vina分析C₄MIM]₃CB与蛋白的结合能,研究溶菌酶与C₄MIM]₃CB相结合的能力。通过亲和萃取和反萃取并进一步脱盐、浓缩、冷冻干燥,得到溶菌酶产品。结果表明,通过对影响因素的研究,优化出最佳的萃取分离条件:蛋清粉溶液pH为7,新型离子液体中C₄MIM]₃CB]浓度为8 mg/mL,反应震荡时间为15 min,反萃取KCl溶液pH为7.0,浓度为1.5 mol/L。最终250 mg蛋清粉可得12.3 mg的溶菌酶产品,纯度为97.56%,酶活力为35000 U/mg,纯化倍数达到37.39,酶活回收率89.74%,且与其他蛋白达到完全分离。该方法具有选择性高、操作简便、产物纯度和活性高等优点。

Affinity Extraction of Lysozyme from Egg White by Novel Ionic Liquid

The method was established and optimized of extracting lysozyme from egg white using the novel ionic liquidC₄MIM]₃CB which was obtained by modifyingC₄MIM]Cl with Cibacron Blue F-3GA,the binding energy betweenC₄MIM]₃CB and protein was analyzed by AutoDock Vina to study the binding ability of lysozyme toC₄MIM]₃CB. Lysozyme products were obtained by affinity extraction and reverse extraction,desalination,concentration and freeze-drying. Results showed that,by studying of the influenced factor,the best extraction and separation conditions for above process were concluded as followed:the optimum pH for egg white powder solution was 7.0 and theC₄MIM]₃CB] containing ionic liquid was at a concentration of 8 mg/mL,the shaking time was 15 min and conditions for back-extracted KCl solution was 1.5 mol/L with pH7.0. Finally,12.3 mg lysozyme with 97.56% purity and 35000 U/mg activity could be obtained by 250 mg egg white power. The purified multiple was 37.39 and the enzyme recovery rate was 89.74%. The method had the advantages of high selectivity,simple operation,high purity and high activity.