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New cationic exchanger support for reversible immobilization of proteins

机译：新型阳离子交换剂支持蛋白质的可逆固定化

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New tailor-made cationic exchange resins have been prepared by covalently binding aspartic-dextran polymers (e.g. MW 15 000-20 000) to porous supports (aminated agarose and Sepabeads). More than 80% of the proteins contained in crude extracts from Escherichia coli and Acetobacter turbidans have been strongly adsorbed on these porous materials at pH 5. This interaction was stronger than in conventional carboxymethyl cellulose (e.g., at pH 7 and 25 degreesC, all proteins previously adsorbed at pH 5 were released from carboxymethyl cellulose, whereas no protein was released from the new supports under similar conditions). Ionic exchange properties of such composites were strongly dependent on the size of the aspartic-dextran polymers as well as on the exact conditions of the covalent coating of the solids with the polymer (optimal conditions: 100 mg aspartic-dextran 20 000/(mL of support); room temperature). Finally, some industrially relevant enzymes (Kluyueromices lactis, Aspergillus oryzae, and Thermus sp. beta-galactosidases, Candida antarctica B lipase, and bovine pancreas trypsin and chymotrypsin) have been immobilized on these supports with very high activity recovery and immobilization rates. After enzyme inactivation, the enzyme can be fully desorbed from the support and the support could be reused for several cycles.
机译：新的量身定制的阳离子交换树脂是通过将天冬-葡聚糖聚合物（例如MW 15000-20000）与多孔载体（胺化的琼脂糖和Sepabeads）共价结合而制备的。在pH 5时，大肠杆菌和浑浊醋杆菌粗提物中所含蛋白质的80％以上已被强烈吸附在这些多孔材料上。这种相互作用比常规的羧甲基纤维素要强（例如，在pH 7和25℃时，所有蛋白质先前在pH 5下吸附的碳会从羧甲基纤维素中释放出来，而在相似的条件下，新载体不会释放蛋白质。此类复合材料的离子交换性能在很大程度上取决于天冬氨酸-葡聚糖聚合物的尺寸以及固体与聚合物共价包被的确切条件（最佳条件：100 mg天冬氨酸-葡聚糖20 000 /（mL支持）;室温）。最后，一些工业相关的酶（乳酸克鲁维酵母，米曲霉和Thermus sp。β-半乳糖苷酶，南极假丝酵母B脂肪酶，牛胰胰蛋白酶和胰凝乳蛋白酶）已被固定在这些载体上，具有很高的活性回收率和固定率。酶灭活后，酶可以从载体上完全解吸，并且载体可以重复使用数个周期。

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