Zhou Xiao-Ming, Entwistle Aiman, Zhang Hong, Jackson Antony P, Mason Thomas O, Shimanovich Ulyana, Knowles Tuomas P J, Smith Andrew T, Sawyer Elizabeth B, Perrett Sarah
National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences 15 Datun Road, Chaoyang District, Beijing 100101 (China) ; Department of Chemistry, University of Cambridge Lensfield Road, Cambridge CB2 1EW (UK) ; University of the Chinese Academy of Sciences 19 A Yuquanlu, Shijingshan District, Beijing 100049 (China).
National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences 15 Datun Road, Chaoyang District, Beijing 100101 (China).
ChemCatChem. 2014 Jul;6(7):1961-1968. doi: 10.1002/cctc.201402125. Epub 2014 Jun 4.
Enzyme immobilization is an important strategy to enhance the stability and recoverability of enzymes and to facilitate the separation of enzymes from reaction products. However, enzyme purification followed by separate chemical steps to allow immobilization on a solid support reduces the efficiency and yield of the active enzyme. Here we describe polypeptide constructs that self-assemble spontaneously into nanofibrils with fused active enzyme subunits displayed on the amyloid fibril surface. We measured the steady-state kinetic parameters for the appended enzymes in situ within fibrils and compare these with the identical protein constructs in solution. Finally, we demonstrated that the fibrils can be recycled and reused in functional assays both in conventional batch processes and in a continuous-flow microreactor.
酶固定化是提高酶的稳定性和可回收性以及促进酶与反应产物分离的重要策略。然而，先进行酶纯化，然后通过单独的化学步骤将其固定在固体载体上，会降低活性酶的效率和产量。在此，我们描述了一种多肽构建体，它能自发地自组装成纳米纤维，融合的活性酶亚基展示在淀粉样纤维表面。我们原位测量了纤维内附加酶的稳态动力学参数，并将其与溶液中相同的蛋白质构建体进行比较。最后，我们证明了这些纤维在传统分批过程和连续流微反应器中的功能测定中都可以循环再利用。
