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酶原在固液界面的吸附与活化。I. 胰凝乳蛋白酶原在烷基氨基修饰的二氧化硅衍生物上的吸附与活化

Adsorption and activation of zymogens at solid-liquid interfaces. I. Chymotrypsinogen on alkylamino modified silica derivatives.

作者信息

Yen H R, Oscarsson S, Ulbrich K, Kopecek J

机构信息

Department of Bioengineering, University of Utah, Salt Lake City 84112.

出版信息

J Biomed Mater Res. 1994 Feb;28(2):247-57. doi: 10.1002/jbm.820280216.

Abstract

Silica beads were modified with alkylamino groups of different lengths (C2, C4, C6, C8, and C10) and hydrophobicity. The relationship between surface structure and adsorption of chymotrypsinogen followed by its activation with trypsin at the solid-liquid interface was studied. From the adsorption isotherms, it follows that underivatized silica adsorbed chymotrypsinogen (CTG) well. The adsorption of CTG on alkylamino modified silicas appeared to correlate with the hydrophobicity of the latter. The longer the alkyl chains were, the higher was the amount of adsorbed CTG. The activation of adsorbed CTG with trypsin at the solid-liquid interface was a slower process when compared with the activation conducted in solution. Parallel experiments were performed with chymotrypsin (CT). The adsorption behavior was similar to that of CTG. The activity of adsorbed CT was inversely proportional to the hydrophobicity of the beads. These results correlated well with the desorption of CT after repeated washings. Repeated addition of substrate (Gly-Gly-Phe-NAp) to the CT covered beads resulted in the CT desorption. The higher the hydrophobicity of the beads was, the lower was the desorption of CT.

摘要

用不同长度(C2、C4、C6、C8和C10)和疏水性的烷基氨基对硅胶珠进行了改性。研究了在固液界面上胰凝乳蛋白酶原的吸附及其随后用胰蛋白酶激活过程中表面结构与吸附之间的关系。从吸附等温线可知,未衍生化的硅胶对胰凝乳蛋白酶原(CTG)有良好的吸附作用。CTG在烷基氨基改性硅胶上的吸附似乎与后者的疏水性相关。烷基链越长,CTG的吸附量越高。与在溶液中进行的激活相比,在固液界面上用胰蛋白酶激活吸附的CTG是一个较慢的过程。用胰凝乳蛋白酶(CT)进行了平行实验。吸附行为与CTG相似。吸附的CT的活性与珠子的疏水性成反比。这些结果与反复洗涤后CT的解吸情况密切相关。向覆盖有CT的珠子中反复添加底物(甘氨酰 - 甘氨酰 - 苯丙氨酸 - N - 乙酰苯胺)导致CT解吸。珠子的疏水性越高,CT的解吸越低。

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