Matter‐tag: A universal immobilization platform for enzymes on polymers, metals, and silicon‐based materials

Publication year: 2020
Authors: Dedisch S. 12, Wiens A. 2, Davari M.D. 2, Söder D. 1, Rodriguez‐Emmenegger C. 13, Jakob F. 12, Schwaneberg U.*12
Affiliations:
1 - DWILeibniz Institute for Interactive Materials, Aachen, Germany
2 - Lehrstuhl für Biotechnologie, RWTH Aachen University, Aachen, Germany
3 - Institute of Technical and Macromolecular Chemistry, RWTH Aachen University, Aachen, Germany
Published in: Biotechnology and Bioengineering, 2020, Vol. 117, Issue1, p. 49-61
doi: 10.1002/bit.27181
Enzymeimmobilization is extensively studied to improve enzymeproperties in catalysis and analytical applications. Here, we introduce a simple and versatile enzymeimmobilization platform based on adhesion-promoting peptides, namely Matter-tags. Matter-tags immobilize enzymes in an oriented way as a dense monolayer. The immobilizationplatform was established with three adhesion-promoting peptides; Cecropin A (CecA), liquid chromatography peak I (LCI), and Tachystatin A2 (TA2), that were genetically fused to enhanced green fluorescentprotein and to two industrially important enzymes: aphytase (from Yersinia mollaretii) and a cellulase (Cel A2 from a metagenomic library). Here, we report a universal and simple Matter-tagbased immobilizationplatform for enzymes on various materials including polymers (polystyrene, polypropylene and polyethyleneterephthalate), metals (stainless steel and gold), and silicon-based materials (siliconwafer). The Matter-tagbased enzyme immobilization is performed at ambient temperature within minutes (<10min) in anaqueous solution harboring the phytase or cellulase by immersing the targeted material. The peptide LCI was identified as universal adhesion promoter; LCI immobilized both enzymes on all investigated materials. The attachment of phytase-LCI onto gold was characterized with surface plasmon resonance spectroscopy obtaining a dissociation constant value (KD) of 2.9·10−M and a maximal surface coverage of 504ng/cm.

MP-SPR keywords: adhesion-promoting peptides, adsorption, binding affinity (KD), enzyme, peptide, surface coverage