Başar Nilgün, Uzun Lokman, Güner Ali, Denizli Adil
Department of Chemistry, Hacettepe University, Ankara, Turkey.
Int J Biol Macromol. 2007 Aug 1;41(3):234-42. doi: 10.1016/j.ijbiomac.2007.02.009. Epub 2007 Mar 2.
Magnetic poly(2-hydroxyethyl methacrylate) mPHEMA beads carrying Cibacron Blue F3GA were prepared by suspension polymerization of HEMA in the presence of Fe3O4 nano-powder. Average size of spherical beads was 80-120 microm. The beads had a specific surface area of 56.0m(2)/g. The characteristic functional groups of dye-attached mPHEMA beads were analyzed by Fourier transform infrared spectrometer (FTIR) and Raman spectrometer. mPHEMA with a swelling ratio of 68% and carrying 28.5 micromol CibacronBlueF3GA/g were used for the purification of lysozyme. Adsorption studies were performed under different conditions in a magnetically stabilized fluidized bed (i.e., pH, protein concentration, flow-rate, temperature, and ionic strength). Lysozyme adsorption capacity of mPHEMA and mPHEMA/Cibacron Blue F3GA beads were 0.8 mg/g and 342 mg/g, respectively. It was observed that after 20 adsorption-desorption cycle, mPHEMA beads can be used without significant loss in lysozyme adsorption capacity. Purification of lysozyme from egg white was also investigated. Purification of lysozyme was monitored by determining the lysozyme activity using Micrococcus lysodeikticus as substrate. The purity of the desorbed lysozyme was about 87.4% with recovery about 79.6%. The specific activity of the desorbed lysozyme was high as 41.586 U/mg.
通过在Fe3O4纳米粉末存在下使甲基丙烯酸2-羟乙酯（HEMA）进行悬浮聚合，制备了负载汽巴克隆蓝F3GA的磁性聚甲基丙烯酸2-羟乙酯（mPHEMA）微珠。球形微珠的平均尺寸为80 - 120微米。这些微珠的比表面积为56.0平方米/克。通过傅里叶变换红外光谱仪（FTIR）和拉曼光谱仪对负载染料的mPHEMA微珠的特征官能团进行了分析。溶胀率为68%且负载28.5微摩尔汽巴克隆蓝F3GA/克的mPHEMA用于溶菌酶的纯化。在磁稳定流化床中于不同条件下（即pH、蛋白质浓度、流速、温度和离子强度）进行吸附研究。mPHEMA和mPHEMA/汽巴克隆蓝F3GA微珠对溶菌酶的吸附容量分别为0.8毫克/克和342毫克/克。观察到经过20次吸附 - 解吸循环后，mPHEMA微珠在溶菌酶吸附容量方面可继续使用而无显著损失。还研究了从蛋清中纯化溶菌酶的情况。以溶壁微球菌为底物通过测定溶菌酶活性来监测溶菌酶的纯化过程。解吸得到的溶菌酶纯度约为87.4%，回收率约为79.6%。解吸得到的溶菌酶的比活性高达41.586 U/毫克。
