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

Abstract

Enzyme immobilization is extensively studied to improve enzyme properties in catalysis and analytical applications. Here, we introduce a simple and versatile enzyme immobilization platform based on adhesion‐promoting peptides, namely Matter‐tags. Matter‐tags immobilize enzymes in an oriented way as a dense monolayer. The immobilization platform 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 fluorescent protein and to two industrially important enzymes: a phytase (from Yersinia mollaretii) and a cellulase (CelA2 from a metagenomic library). Here, we report a universal and simple Matter‐tag–based immobilization platform for enzymes on various materials including polymers (polystyrene, polypropylene, and polyethylene terephthalate), metals (stainless steel and gold), and silicon‐based materials (silicon wafer). The Matter‐tag–based enzyme immobilization is performed at ambient temperature within minutes (<10 min) in an aqueous 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 2.9·10−8 M and a maximal surface coverage of 504 ng/cm².

Countercurrently Operated Reactive Extractor with an Additively Manufactured Enzyme Carrier Structure

“…Immobilisates of this type can be used for the synthesis of MVP from FA s in a buffer-heptane two-phase system. 23 The universal applicability of one anchor to different materials with similar properties has been shown by Dedisch et al 24 . Manufacturing POCS out of PET with fused deposition modelling (FDM) is limited by the resolution and effects like string formation between unit cells during the manufacturing process are affecting the optimized flow of heptane.…”

A peptide‐based coating toolbox to enable click chemistry on polymers, metals, and silicon through sortagging

“…In deviation from the previous protocol, the reaction time was increased to 18 h to ensure maximum conversion of amine compounds. Subsequently, , 2020Meurer et al, 2017;Rübsam et al, 2017).…”

“…APs can be used in a resource-and energy-efficient manner for highly dense and specific surface functionalization at ambient temperatures in water and represent an attractive alternative to plasma treatment, UV irradiation, and employment of strong acids/bases (Dedisch et al, 2020;Goddard & Hotchkiss, 2007;Li et al, 2004;Rübsam et al, 2017;Vesel & Mozetic, 2017). The AP liquid chromatography peak I (LCI, 47 amino acids) from Bacillus subtilis was identified as a universal material binding peptide for functionalization of polymer surfaces (e.g., polypropylene [PP] and polyethylene terephthalate [PET]; Dedisch et al, 2020;Rübsam et al, 2017), metals (e.g., stainless steel and gold; Dedisch et al, 2020), silicon-based materials (Dedisch et al, 2020), and natural surfaces (e.g., teeth (Dedisch et al, 2019), hair (Dedisch et al, 2019), and plant leaves (Meurer et al, 2017)). LCI shows significantly superior binding strength to the latter surfaces when compared to proteins such as enhanced green fluorescent protein (EGFP), phytase, cellulase, and bovine serum albumin (Dedisch et al, 2019(Dedisch et al, , 2020Meurer et al, 2017;Rübsam et al, 2017).…”

“…Anchor peptides (APs) were applied as adhesion promotors to immobilize functional moieties such as enzymes (Zernia et al, 2016), bioactive peptides (Liu et al, 2016), antigens (de Juan‐Franco et al, 2013), containers for drug delivery (Apitius et al, 2019), synthetic polymers (Dedisch et al, 2019; Meurer et al, 2017), and organometallic catalysts (Grimm et al, 2019) through noncovalent interactions. APs can be used in a resource‐ and energy‐efficient manner for highly dense and specific surface functionalization at ambient temperatures in water and represent an attractive alternative to plasma treatment, UV irradiation, and employment of strong acids/bases (Dedisch et al, 2020; Goddard & Hotchkiss, 2007; Li et al, 2004; Rübsam et al, 2017; Vesel & Mozetic, 2017). The AP liquid chromatography peak I (LCI, 47 amino acids) from Bacillus subtilis was identified as a universal material binding peptide for functionalization of polymer surfaces (e.g., polypropylene [PP] and polyethylene terephthalate [PET]; Dedisch et al, 2020; Rübsam et al, 2017), metals (e.g., stainless steel and gold; Dedisch et al, 2020), silicon‐based materials (Dedisch et al, 2020), and natural surfaces (e.g., teeth (Dedisch et al, 2019), hair (Dedisch et al, 2019), and plant leaves (Meurer et al, 2017)).…”

Tunable Polymeric Scaffolds for Enzyme Immobilization

“…Furthermore, natural or artificial epitopes or tags are strategically inserted in the protein in order to promote a favorable orientation, which usually enhances the catalytic performance of the enzyme. Several examples of peptide-tags that are artificially introduced in recombinant proteins can be found in the literature [e.g., cellulose-binding domains (CBDs) ( Dai et al, 2017 ), SNAP Tag ( Fang et al, 2019 ), matter-tag ( Dedisch et al, 2020 ), solid-binding peptides ( Care et al, 2017 ), or FLAG tag ( Vishwanath et al, 1997 )]. The most used polymers bear affinity tags such as nitrilotriacetic acid (NTA) functionalization or avidin/streptavidin motifs, which are used to tether His-tagged or biotinylated proteins, respectively.…”