Immobilization of 2-Deoxy-d-ribose-5-phosphate Aldolase in Polymeric Thin Films via the Langmuir-Schaefer Technique

Stefan Reinicke; Huw C Rees; Pieter Espeel; Nane Vanparijs; Carolin Bisterfeld; Markus Dick; Ruben R Rosencrantz; Gerald Brezesinski; Bruno G de Geest; Filip E Du Prez; Jörg Pietruszka; Alexander Böker

doi:10.1021/acsami.6b13632

Immobilization of 2-Deoxy-d-ribose-5-phosphate Aldolase in Polymeric Thin Films via the Langmuir-Schaefer Technique

ACS Appl Mater Interfaces. 2017 Mar 8;9(9):8317-8326. doi: 10.1021/acsami.6b13632. Epub 2017 Feb 21.

Authors

Stefan Reinicke¹, Huw C Rees², Pieter Espeel³, Nane Vanparijs⁴, Carolin Bisterfeld⁵, Markus Dick⁵, Ruben R Rosencrantz^{1

6}, Gerald Brezesinski⁷, Bruno G de Geest⁴, Filip E Du Prez³, Jörg Pietruszka^{5

8}, Alexander Böker^{1

6}

Affiliations

¹ Department of Functional Protein Systems and Biotechnology, Fraunhofer Institute of Applied Polymer Research (IAP) , Geiselbergstraße 69, 14476, Potsdam-Golm, Germany.
² Department of Chemistry, University of Chicago , Chicago, Illinois 60637, United States.
³ Department of Organic and Macromolecular Chemistry, Polymer Chemistry Research Group, Ghent University , Krijgslaan 281 S4-bis, 9000 Ghent, Belgium.
⁴ Department of Pharmaceutics, Ghent University , Ottergemsesteenweg 460, 9000 Ghent, Belgium.
⁵ Institut of Bioorganic Chemistry, Heinrich Heine University of Düsseldorf at Forschungszentrum Jülich , Stetternicher Forst, D-52426 Jülich, Germany.
⁶ Polymer Materials and Polymer Technologies, University of Potsdam , 14476, Potsdam-Golm, Germany.
⁷ Max Planck Institute of Colloids and Interfaces , Science Park Potsdam-Golm, Am Mühlenberg 1, 14476 Potsdam, Germany.
⁸ IBG-1: Biotechnology, Forschungszentrum Jülich GmbH , 52425 Jülich, Germany.

PMID: 28186396
DOI: 10.1021/acsami.6b13632

Abstract

A synthetic protocol for the fabrication of ultrathin polymeric films containing the enzyme 2-deoxy-d-ribose-5-phosphate aldolase from Escherichia coli (DERA_EC) 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.

Keywords: 2-deoxy-d-ribose-5-phosphate aldolase; Langmuir−Schaefer; enzyme immobilization; poly(N-isopropylacrylamide); polymeric thin film; thiolactone.