Cross-linked enzyme aggregates (CLEAs): A novel and versatile method for enzyme immobilization (a review)
Cross-linked enzyme aggregates (CLEAs): A novel and versatile method for enzyme immobilization (a review)
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DOI：
10.1080/10242420500183378
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发表时间：
2005-01
影响因子：
1.8
通讯作者：
R. Sheldon;R. Schoevaart;L. V. Langen
中科院分区：
文献类型：
--
作者：
R. Sheldon;R. Schoevaart;L. V. Langen
获得酶最佳性能的关键往往是设计一种有效的固定化方法。本文综述了一种新型的、通用的、有效的酶固定化方法，即交联酶聚集体（CLEA）。该方法非常简单-包括从水性缓冲液中沉淀酶，然后交联酶分子的物理聚集体-并且易于快速优化。它适用于多种酶，包括辅因子依赖性氧化还原酶和裂解酶，并提供稳定的、可回收的催化剂，具有高活性保留，有时高于其衍生的游离酶的活性。酶不需要高纯度。实际上，该方法基本上是纯化和固定在一个步骤中的组合。该技术也适用于制备含有两种或更多种酶的combi-CLEA，用于一锅法、多步合成。例如，使用醇腈酶/腈水解酶combi-CLEA以高产率和对映体纯度从苯甲醛一锅法合成（S）-扁桃酸。
The key to obtaining optimum performance of an enzyme is often a question of devising an effective method for its immobilization. This review describes a novel, versatile and effective methodology for enzyme immobilization, namely, as cross-linked enzyme aggregates (CLEAs). The method is exquisitely simple – involving precipitation of the enzyme from aqueous buffer followed by cross-linking of the resulting physical aggregates of enzyme molecules – and amenable to rapid optimization. It is applicable to a wide variety of enzymes, including cofactor-dependent oxidoreductases and lyases, and affords stable, recyclable catalysts with high retention of activity, sometimes higher than that of the free enzyme it was derived from. The enzyme does not need to be of high purity. Indeed, the methodology is essentially a combination of purification and immobilization in one step. The technique is also applicable to the preparation of combi-CLEAs, containing two or more enzymes, for use in one-pot, multi-step syntheses. For example, an oxynitrilase/nitrilase combi-CLEA was used for the one-pot synthesis of (S)-mandelic acid from benzaldehyde, in high yield and enantiomeric purity.