Immobilization of Proteins on Highly Activated Glyoxyl Supports: Dramatic Increase of the Enzyme Stability via Multipoint Immobilization on Pre-existing Carriers
Immobilization of Proteins on Highly Activated Glyoxyl Supports: Dramatic Increase of the Enzyme Stability via Multipoint Immobilization on Pre-existing Carriers
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DOI：
10.2174/138527281917150806125708
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发表时间：
2015-01-01
影响因子：
2.6
通讯作者：
Guisan, Jose M.
中科院分区：
文献类型：
--
作者：
Fernandez-Lorente, Gloria;Lopez-Gallego, Fernando;Guisan, Jose M.
将酶与乙氧基结合在活性高的载体上是一种通过多点共价结合在已有载体上来提高酶稳定性的极好方法。乙氧基是通过甘油基团(载体-O-CH2-CHOH-CH2OH)的高碘酸盐氧化得到的短链脂肪醛(载体-O-CH2-CHO)。乙氧基基团的独特特征如下：a.-通过其氨基对酶的固定化必须通过多点连接发生。b.-乙氧基基团在pH 10下稳定，这允许赖氨酸参与固定化过程。c.乙氧基基团在pH 10下非常稳定，这允许固定化酶和活化载体之间的长期孵育，以促进非常强烈的酶-载体多点共价连接。使用该方案，100多种工业酶高度稳定。在许多情况下，获得了比传统方法产生的固定化衍生物稳定1000倍以上的稳定性。虽然固定化酶具有很好的稳定性，但其催化活性仅为相应的可溶酶的50%~90%。工业酶的稳定是固定化方案中的关键步骤。酶被固定化，以供工业规模使用一系列反应周期。
The binding of enzymes on carriers with a high degree of activation with glyoxyl groups is an excellent method for improving enzyme stability by multipoint covalent attachment on a pre-existing carrier. Glyoxyl groups are short aliphatic aldehyde groups (Support-O-CH2-CHO) that can be obtained by periodate oxidation of glyceryl groups (Support-O-CH2-CHOH-CH2OH). The unique features of glyoxyl groups are as follows:a.-The immobilization of enzymes through their amino groups has to occur via multipoint attachment.b.-The glyoxyl groups are stable at pH 10, which allows for the participation of Lys in the immobilization process.c.-The glyoxyl groups are very stable at pH 10, which allows for a long-term incubation between the immobilized enzyme and the activated support to promote a very intense enzyme-support multipoint covalent attachment.Using this protocol, more than 100 industrial enzymes were highly stabilized. In many cases, stabilizations of greater than 1000-fold compared with immobilized derivatives generated by conventional methods were obtained. Although dramatic stabilization was achieved, the immobilized enzymes maintained only 50 to 90 % of the catalytic activity of the corresponding soluble enzyme. Stabilization of industrial enzymes is a key step in immobilization protocols. Enzymes are immobilized for use at industrial scales for a number of reaction cycles.