Use of reversible denaturation for adsorptive immobilization of urease
Use of reversible denaturation for adsorptive immobilization of urease
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DOI：
10.1385/abab:94:3:265
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发表时间：
2001-06-01
影响因子：
3
通讯作者：
Nemat-Gorgani, M
中科院分区：
文献类型：
--
作者：
Azari, F;Hosseinkhani, S;Nemat-Gorgani, M
选择尿素酶作为模型多聚体蛋白，研究可逆变性用于固定到疏水支持物的效用。在研究的各种程序中，酸变性提供了最高程度的固定化和酶活性，同时降低了Km（表观）。通过使用8-苯胺基-1-萘磺酸盐作为疏水报告探针的荧光研究证实了由酸性pH环境诱导的蛋白质分子中疏水簇的暴露。酶在低pH值下的催化潜力显着提高，显着的热和pH稳定性增强固定化。此外，固定化制剂被成功地用于连续催化转化。基于本文中提出的结果和最近的一份报告，涉及一个相对更简单的单体蛋白质，它建议，可逆变性可能是一般实用程序的蛋白质，通常不吸附在疏水支持物上的固定化。
Urease was chosen as a model multimeric protein to investigate the utility of reversible denaturation for immobilization to a hydrophobic support. Of the various procedures investigated, acidic denaturation provided the highest degree of immobilization and enzymatic activity with lowering of K-m (apparent). Exposure of hydrophobic clusters in the protein molecule induced by the acidic pH environment was confirmed by fluorescence studies using 8-anilino-1-naphtalene-sulfonate as a hydrophobic-reporter probe. The catalytic potential of the enzyme at low pH values was dramatically improved with significant heat and pH stability enhancement on immobilization. Furthermore, the immobilized preparation was used successfully in continuous catalytic transformations. Based on the results presented in this article and a recent report involving a relatively more simple monomeric protein, it is suggested that reversible denaturation may be of general utility for immobilization of proteins, which are not normally adsorbed on hydrophobic supports.