Department of Functional Protein Systems and Biotechnology, Fraunhofer Institute for Applied Polymer Research (IAP) , Geiselbergstraße 69, 14476 Potsdam-Golm, Germany.
Polymer Materials and Polymer Technologies, University of Potsdam , 14476, Potsdam-Golm, Germany.
Bioconjug Chem. 2018 Jan 17;29(1):104-116. doi: 10.1021/acs.bioconjchem.7b00645. Epub 2017 Dec 13.
2-Deoxy-d-ribose-5-phosphate aldolase (DERA) is a biocatalyst that is capable of converting acetaldehyde and a second aldehyde as acceptor into enantiomerically pure mono- and diyhydroxyaldehydes, which are important structural motifs in a number of pharmaceutically active compounds. However, substrate as well as product inhibition requires a more-sophisticated process design for the synthesis of these motifs. One way to do so is to the couple aldehyde conversion with transport processes, which, in turn, would require an immobilization of the enzyme within a thin film that can be deposited on a membrane support. Consequently, we developed a fabrication process for such films that is based on the formation of DERA-poly(N-isopropylacrylamide) conjugates that are subsequently allowed to self-assemble at an air-water interface to yield the respective film. In this contribution, we discuss the conjugation conditions, investigate the interfacial properties of the conjugates, and, finally, demonstrate a successful film formation under the preservation of enzymatic activity.
2-脱氧-D-核糖-5-磷酸醛缩酶(DERA)是一种生物催化剂,能够将乙醛和第二种醛作为受体转化为对映体纯的单羟醛和二羟醛,这些都是许多具有药用活性的化合物中的重要结构基序。然而,底物和产物抑制需要更复杂的工艺设计来合成这些基序。一种方法是将醛转化与传输过程相结合,这反过来又需要将酶固定在可以沉积在膜支撑物上的薄膜内。因此,我们开发了一种基于 DERA-聚(N-异丙基丙烯酰胺)缀合物形成的制造此类薄膜的方法,随后允许其在气-水界面处自组装以得到相应的薄膜。在本研究中,我们讨论了缀合条件,研究了缀合物的界面性质,并最终证明了在保持酶活性的情况下成功形成了薄膜。