School of Biological Sciences, University of Canterbury, Christchurch, New Zealand.
Biotechnol Prog. 2010 Jan-Feb;26(1):93-100. doi: 10.1002/btpr.309.
Amyloid fibrils are a misfolded state, formed by many proteins when subjected to denaturing conditions. Their constituent amino acids make them ideally suited as a readily functionalized nanoscaffold for enzyme immobilization and their strength, stability, and nanometer size are attractive features for exploitation in the creation of new bionanomaterials. We report successful functionalization of amyloid fibrils by conjugation to glucose oxidase (GOD) using glutaraldehyde. GOD retained activity upon attachment and successful cross-linking was determined using electrophoresis, centrifugation, sucrose gradient centrifugation, and TEM. The resulting functionalized enzyme scaffold was then incorporated into a model poly(vinyl alcohol) (PVOH) film, to create a new bionanomaterial. The antibacterial effect of the functionalized film was then tested on E. coli, the growth of which was inhibited, demonstrating the incorporation of GOD antibacterial activity into the PVOH film. The incorporation of the GOD-functionalized amyloid fibrils into PVOH provides an excellent 'proof of concept' model for the creation of a new bionanomaterial using a functionalized amyloid fibril scaffold.
淀粉样纤维是一种错误折叠的状态，由许多蛋白质在变性条件下形成。其组成氨基酸使它们成为酶固定化的理想功能化纳米支架，其强度、稳定性和纳米尺寸是开发新型生物纳米材料的有吸引力的特征。我们报告了通过使用戊二醛将葡萄糖氧化酶（GOD）缀合到淀粉样纤维上来实现成功的功能化。在附着时保留了 GOD 的活性，并通过电泳、离心、蔗糖梯度离心和 TEM 确定了成功的交联。然后将所得功能化酶支架掺入模型聚乙烯醇（PVOH）膜中，以创建新的生物纳米材料。然后在大肠杆菌上测试功能化膜的抑菌效果，其生长受到抑制，表明 GOD 的抑菌活性已掺入 PVOH 膜中。将 GOD 功能化的淀粉样纤维掺入 PVOH 中为使用功能化淀粉样纤维支架创建新的生物纳米材料提供了极好的“概念验证”模型。
