Bayramoglu Gulay, Tekinay Turgay, Ozalp V Cengiz, Arica M Yakup
Biochemical Processing and Biomaterial Research Laboratory, Gazi University, 06500 Teknikokullar, Ankara, Turkey; Department of Chemistry, Faculty of Sciences, Gazi University, 06500 Teknikokullar, Ankara, Turkey.
Gazi University, Life Sciences Application and Research Center, Golbasi, 06830 Ankara, Turkey.
J Chromatogr B Analyt Technol Biomed Life Sci. 2015 May 15;990:84-95. doi: 10.1016/j.jchromb.2015.03.030. Epub 2015 Apr 2.
Lysozyme is an important polypetide used in medical and food applications. We report a novel magnetic strong cation exchange beads for efficient purification of lysozyme from chicken egg white. Magnetic chitosan (MCHT) beads were synthesized via phase inversion method, and then grafted with poly(glycidyl methacrylate) (p(GMA)) via the surface-initiated atom transfer radical polymerization (SI-ATRP). Epoxy groups of the grafted polymer, were modified into strong cation-exchange groups (i.e., sulfonate groups) in the presence of sodium sulfite. The MCTH and MCTH-g-p(GMA)-SO3H beads were characterized by ATR-FTIR, SEM, and VSM. The sulphonate groups content of the modified MCTH-g-p(GMA)-4 beads was found to be 0.53mmolg(-1) of beads by the potentiometric titration method. The MCTH-g-p(GMA)-SO3H beads were first used as an ion-exchange support for adsorption of lysozyme from aqueous solution. The influence of different experimental parameters such as pH, contact time, and temperature on the adsorption process was evaluated. The maximum adsorption capacity was found to be 208.7mgg(-1) beads. Adsorption of lysozyme on the MCTH-g-p(GMA)-SO3H beads fitted to Langmuir isotherm model and followed the pseudo second-order kinetic. More than 93% of the adsorbed lysozyme was desorbed using Na2CO3 solution (pH 11.0). The purity of the lysozyme was checked by HPLC and SDS gel electrophoresis. In addition, the MCTH-g-p(GMA)-SO3H beads prepared in this work showed promising potential for separation of various anionic molecules.
溶菌酶是一种用于医学和食品应用的重要多肽。我们报道了一种新型磁性强阳离子交换珠，用于从鸡蛋白中高效纯化溶菌酶。通过相转化法合成了磁性壳聚糖（MCHT）珠，然后通过表面引发原子转移自由基聚合（SI-ATRP）接枝聚（甲基丙烯酸缩水甘油酯）（p（GMA））。接枝聚合物的环氧基团在亚硫酸钠存在下被改性为强阳离子交换基团（即磺酸基团）。通过ATR-FTIR、SEM和VSM对MCTH和MCTH-g-p（GMA）-SO3H珠进行了表征。通过电位滴定法发现改性的MCTH-g-p（GMA）-4珠的磺酸基团含量为0.53mmol g（-1）珠。MCTH-g-p（GMA）-SO3H珠首先用作离子交换载体，用于从水溶液中吸附溶菌酶。评估了不同实验参数如pH、接触时间和温度对吸附过程的影响。发现最大吸附容量为208.7mg g（-1）珠。溶菌酶在MCTH-g-p（GMA）-SO3H珠上的吸附符合朗缪尔等温线模型，并遵循准二级动力学。使用Na2CO3溶液（pH 11.0）解吸了超过93%的吸附溶菌酶。通过HPLC和SDS凝胶电泳检查了溶菌酶的纯度。此外，本工作制备的MCTH-g-p（GMA）-SO3H珠在分离各种阴离子分子方面显示出有前景的潜力。
