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首页> 外文期刊>Biochemical Engineering Journal >Immobilization of Bacillus circulans β-galactosidase and its application in the synthesis of galacto-oligosaccharides under repeated-batch operation
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Immobilization of Bacillus circulans β-galactosidase and its application in the synthesis of galacto-oligosaccharides under repeated-batch operation

机译：反复分批操作的环状芽孢杆菌β-半乳糖苷酶的固定化及其在合成低聚半乳糖中的应用

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摘要

A highly active and stable derivate of immobilized Bacillus circulans β-galactosidase was prepared for the synthesis of galacto-oligosaccharides (COS) under repeated-batch operation. B. circulans fi-galactosidase was immobilized on monofunctional glyoxyl agarose and three heterofunctional supports: amino-, carboxy-, and chelate-glyoxyl agarose. Glyoxyl agarose was the support with highest immobilization yield and stability being selected for the optimization of immobilization conditions and application in GOS synthesis. A central composite rotatable design was conducted to optimize contacted protein and immobilization time, using maximum catalytic potential as the objective function. Optimal conditions of immobilization were 28.9 mg/g and 36.4 h of contact, resulting in a biocatalyst with 595 IU/g and a half-life 89-fold higher than soluble enzyme. Immobilization process did not alter the synthetic capacity of (3-galactosidase, obtaining the same GOS yield and product profile than the free enzyme. GOS yield and productivity remained unchanged along 10 repeated batches, with values of 39% (w/w) and 5.7 g GOS/g of biocatalystbatch. Total product obtained after 10 batches of reaction was 56.5 g GOS/g of biocatalyst (1956 g GOS/g protein). Cumulative productivity in terms of mass of contacted protein was higher for the immobilized enzyme than for its soluble counterpart from the second batch of synthesis onwards.
机译：制备了高活性和稳定的环化芽孢杆菌β-半乳糖苷酶衍生物，用于在分批操作下合成半乳糖寡糖（COS）。圆环芽孢杆菌的半乳糖苷酶固定在单功能乙醛酸琼脂糖和三种杂功能支持物上：氨基，羧基和螯合乙醛酸琼脂糖。乙醛琼脂糖是固定化产率和稳定性最高的载体，其被选择用于固定化条件的优化和在GOS合成中的应用。使用最大的催化潜力作为目标函数，进行了中央复合材料可旋转设计，以优化接触的蛋白质和固定时间。固定化的最佳条件是28.9 mg / g和36.4 h的接触，导致生物催化剂的含量为595 IU / g，半衰期比可溶性酶高89倍。固定化过程没有改变（3-半乳糖苷酶）的合成能力，获得了与游离酶相同的GOS产量和产物谱。在10个重复批次中，GOS产量和生产率保持不变，分别为39％（w / w）和5.7 gGOS / g生物催化剂批处理10批反应后获得的总产物为56.5 g GOS / g生物催化剂（1956 g GOS / g蛋白质）。固定化酶的接触蛋白质质量累计生产率高于其固定化酶从第二批合成开始是可溶的。

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