Boivin P, Kobos R K, Papa S L, Scouten W H
Department of Chemistry, Baylor University, Waco, Texas 76798.
Biotechnol Appl Biochem. 1991 Oct;14(2):155-69.
Perflex has been introduced by E. I. du Pont de Nemours and Co., Inc., as a new fluorocarbon-based technology for protein immobilization. Due to the hydrophobic character of the support, however, significant loss of enzymatic activity may occur upon immobilization of certain enzymes, which appears to be due to a large conformational change of the protein ("inversion"). Pretreatment of the Perflex support with a neutral fluorosurfactant lessened the surface hydrophobicity, thus decreasing the hydrophobic interaction between the support and the protein. Modification of enzymes with a high number of fluorocarbon residues, which forms a hydrophobic "envelope" around the protein, also appears to prevent enzyme inactivation upon immobilization on Perflex support. Moreover, preactivation of the support with either perfluorooctylpropylisocyanate or reactive poly(fluoroalkyl) sugar reagents greatly improves the enzyme particle activity by increasing the amount of immobilized enzyme. Fluorosurfactant treatment of the support activated with perfluorooctylpropylisocyanate improves the retention of activity for sensitive enzymes such as alpha-chymotrypsin and increases the wetability and ease of handling of the Perflex particles.
E. I. 杜邦公司推出了Perflex，这是一种基于氟碳化合物的新型蛋白质固定化技术。然而，由于载体的疏水性，某些酶固定化时可能会出现显著的酶活性损失，这似乎是由于蛋白质发生了较大的构象变化（“反转”）。用中性含氟表面活性剂对Perflex载体进行预处理可降低表面疏水性，从而减少载体与蛋白质之间的疏水相互作用。用大量含氟碳残基修饰酶，这些残基会在蛋白质周围形成疏水“包膜”，这似乎也能防止酶固定在Perflex载体上时失活。此外，用全氟辛基丙基异氰酸酯或反应性聚（氟烷基）糖试剂对载体进行预活化，可通过增加固定化酶的量大大提高酶颗粒活性。用全氟辛基丙基异氰酸酯活化的载体经含氟表面活性剂处理后，可提高诸如α-胰凝乳蛋白酶等敏感酶的活性保留率，并增加Perflex颗粒的润湿性和易操作性。
