Immobilization of Proteins on Highly Activated Glyoxyl Supports: Dramatic Increase of the Enzyme Stability via Multipoint Immobilization on Pre-existing Carriers

Title:Immobilization of Proteins on Highly Activated Glyoxyl Supports: Dramatic Increase of the Enzyme Stability via Multipoint Immobilization on Pre-existing Carriers

Volume: 19 Issue: 17

Author(s): Gloria Fernández-Lorente, Fernando Lopez-Gallego, Juan M Bolivar, Javier Rocha-Martin, Sonia Moreno-Perez and Jose M. Guisan

Affiliation:

Keywords: Immobilization-stabilization of enzymes, enzyme-support multipoint covalent attachment, glyoxyl-agarose supports.

Abstract: 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.

Cite this article as:

Fernández-Lorente Gloria, Lopez-Gallego Fernando, Bolivar M Juan, Rocha-Martin Javier, Moreno-Perez Sonia and Guisan M. Jose, Immobilization of Proteins on Highly Activated Glyoxyl Supports: Dramatic Increase of the Enzyme Stability via Multipoint Immobilization on Pre-existing Carriers, Current Organic Chemistry 2015; 19 (17) . https://dx.doi.org/10.2174/138527281917150806125708