Departamento de Biocatálisis, Instituto de Catálisis-CSIC, Campus UAM-CSIC Madrid, Spain.
Departamento de Biocatálisis, Instituto de Catálisis-CSIC, Campus UAM-CSIC Madrid, Spain; Transformation and Food Product Elaboration Laboratory, Nutrition and Food, Technology Institute (INATAA), University of Brothers Mentouri Constantine 1, Algeria.
J Biotechnol. 2021 Mar 10;329:128-142. doi: 10.1016/j.jbiotec.2021.02.005. Epub 2021 Feb 15.
Trypsin, chymotrypsin, penicillin G acylase and ficin extract have been stabilized by immobilization on glyoxyl agarose, adding different aliphatic compounds bearing a primary amine group during the immobilization: ethyl amine, butyl amine, hexyl amine (at concentrations ranging from 0 to 20 mM) and octyl amine (from 0 to 10 mM) to analyze their effects on the immobilized enzyme stability. As expected, the presence of amines reduced the intensity of the enzyme-support multipoint covalent attachment, and therefore the enzyme stability. However, it is clear that this effect is higher using octyl amine for all enzymes (in some cases the enzyme immobilized in the presence of 10 mM octyl amine was almost inactivated while the reference kept over 50 % of the initial activity). This way, it seems that the most important effect of the presence of aminated compounds came from the generation of steric hindrances to the enzyme/support multi-reaction promoted by the ammines that are interacting with the aldehyde groups. In some instances, just 1 mM of aminated compounds is enough to greatly decrease enzyme stability. The results suggested that, if the composition of the enzyme extract is unknown, to eliminate small aminated compounds may be necessary to maximize the enzyme-support reaction.
胰蛋白酶、糜蛋白酶、青霉素 G 酰化酶和无花果蛋白酶提取物已通过固定在乙二醛琼脂糖上得到稳定，在固定化过程中添加了不同带有伯胺基团的脂肪族化合物：乙胺、丁胺、己胺（浓度范围从 0 到 20mM）和辛胺（0 到 10mM），以分析它们对固定化酶稳定性的影响。正如预期的那样，胺的存在降低了酶-载体多点共价附着的强度，从而降低了酶的稳定性。然而，很明显，对于所有酶来说，使用辛胺的效果更高（在某些情况下，在存在 10mM 辛胺的情况下固定化的酶几乎失活，而参考酶保持超过初始活性的 50%）。这样，似乎带氨基化合物的存在所产生的最重要影响来自于与醛基相互作用的氨对酶/载体的多反应的空间位阻的产生。在某些情况下，仅仅 1mM 的带氨基化合物就足以大大降低酶的稳定性。结果表明，如果不知道酶提取物的成分，为了最大限度地提高酶-载体反应，可能需要去除小的带氨基化合物。
