Department of Functional Protein Systems and Biotechnology, Fraunhofer Institute of Applied Polymer Research (IAP) , Geiselbergstraße 69, 14476, Potsdam-Golm, Germany.
Department of Chemistry, University of Chicago , Chicago, Illinois 60637, United States.
ACS Appl Mater Interfaces. 2017 Mar 8;9(9):8317-8326. doi: 10.1021/acsami.6b13632. Epub 2017 Feb 21.
A synthetic protocol for the fabrication of ultrathin polymeric films containing the enzyme 2-deoxy-d-ribose-5-phosphate aldolase from Escherichia coli (DERA) is presented. Ultrathin enzymatically active films are useful for applications in which only small quantities of active material are needed and at the same time quick response and contact times without diffusion limitation are wanted. We show how DERA as an exemplary enzyme can be immobilized in a thin polymer layer at the air-water interface and transferred to a suitable support by the Langmuir-Schaefer technique under full conservation of enzymatic activity. The polymer in use is a poly(N-isopropylacrylamide-co-N-2-thiolactone acrylamide) (P(NIPAAm-co-TlaAm)) statistical copolymer in which the thiolactone units serve a multitude of purposes including hydrophobization of the polymer, covalent binding of the enzyme and the support and finally cross-linking of the polymer matrix. The application of this type of polymer keeps the whole approach simple as additional cocomponents such as cross-linkers are avoided.
提出了一种用于制备含有来自大肠杆菌的酶 2-脱氧-D-核糖-5-磷酸醛缩酶（DERA）的超薄聚合物膜的合成方案。超薄的酶活性膜在仅需要少量活性物质的应用中非常有用，同时还需要快速响应和接触时间而没有扩散限制。我们展示了如何将 DERA 作为一个典型的酶在空气-水界面的薄聚合物层中固定，并通过 Langmuir-Schäfer 技术在完全保持酶活性的情况下将其转移到合适的载体上。所使用的聚合物是一种聚（N-异丙基丙烯酰胺-co-N-2-硫代内酰胺丙烯酰胺）（P（NIPAAm-co-TlaAm））统计共聚物，其中硫代内酯单元具有多种用途，包括聚合物的疏水化、酶和载体的共价结合以及聚合物基质的交联。这种类型的聚合物的应用使整个方法保持简单，因为避免了额外的共成分，如交联剂。
