Chemistry Department, Adnan Menderes University, Aydın, Turkey.
Appl Biochem Biotechnol. 2013 Aug;170(8):1909-21. doi: 10.1007/s12010-013-0322-z. Epub 2013 Jun 19.
In this study, concanavalin A (Con A)-attached poly(ethylene glycol dimethacrylate) [poly(EGDMA)] cryogels were used for immobilization of Aspergillus niger inulinase. For this purposes, the monolithic cryogel column was prepared by radical cryocopolymerization of EGDMA as a monomer and N,N'-methylene bisacrylamide as a crosslinker. Then, Con A was attached by covalent binding onto amino-activated poly(EGDMA) cryogel via glutaraldehyde activation. Characterization of cryogels was performed by FTIR, EDX, and SEM studies. Poly(EGDMA) cryogels were highly porous and pore size was found to be approximately 50-100 μm. Con A-attached poly(EGDMA) cryogels was used in the adsorption of inulinase from aqueous solutions. Adsorption of inulinase on the Con A-attached poly(EGDMA) cryogel was performed in continuous system and the effects of pH, inulinase concentration, and flow rate on adsorption were investigated. The maximum amount of inulinase adsorption was calculated to be 27.85 mg/g cryogel at 1.0 mg/mL inulinase concentration and in acetate buffer at pH 4.0. Immobilized inulinase was effectively used in continuous preparation of high-fructose syrup. Inulin was converted to fructose in a continuous system and released fructose concentration was found to be 0.23 mg/mL at the end of 5 min of hydrolysis. High-fructose content of the syrup was demonstrated by thin layer chromatography.
在这项研究中，刀豆球蛋白 A（Con A）接枝聚乙二醇二甲基丙烯酸酯 [聚（EGDMA）] 水凝胶被用于固定黑曲霉菊粉酶。为此，通过 EGDMA 作为单体和 N,N'-亚甲基双丙烯酰胺作为交联剂的自由基冷冻共聚制备了整体式 cryogel 柱。然后，通过戊二醛活化，将 Con A 通过共价键结合到氨基活化的聚（EGDMA）cryogel 上。通过 FTIR、EDX 和 SEM 研究对 cryogels 进行了表征。聚（EGDMA）cryogels 具有高度多孔性，孔径约为 50-100 μm。Con A 接枝聚（EGDMA）cryogels 用于从水溶液中吸附菊粉酶。在连续体系中进行了 Con A 接枝聚（EGDMA）cryogel 对菊粉酶的吸附，研究了 pH、菊粉酶浓度和流速对吸附的影响。在 1.0 mg/mL 菊粉酶浓度和 pH 4.0 的乙酸盐缓冲液中，计算出菊粉酶的最大吸附量为 27.85 mg/g cryogel。固定化菊粉酶有效地用于连续制备高果糖浆。在连续体系中，菊粉转化为果糖，水解 5 分钟后，释放的果糖浓度达到 0.23 mg/mL。通过薄层层析证明了糖浆中的高果糖含量。
