Active surfaces engineered by immobilizing protein-polymer nanoreactors for selectively detecting sugar alcohols

Abstract

We introduce active surfaces generated by immobilizing protein-polymer nanoreactors on a solid support for sensitive sugar alcohols detection. First, such selective nanoreactors were engineered in solution by simultaneous encapsulation of specific enzymes in copolymer polymersomes, and insertion of membrane proteins for selective conduct of sugar alcohols. Despite the artificial surroundings, and the thickness of the copolymer membrane, functionality of reconstituted Escherichia coli glycerol facilitator (GlpF) was preserved, and allowed selective diffusion of sugar alcohols to the inner cavity of the polymersome, where encapsulated ribitol dehydrogenase (RDH) enzymes served as biosensing entities. Ribitol, selected as a model sugar alcohol, was detected quantitatively by the RDH-nanoreactors with GlpF-mediated permeability in a concentration range of 1.5-9 mM. To obtain "active surfaces" for detecting sugar alcohols, the nanoreactors optimized in solution were then immobilized on a solid support: aldehyde groups exposed at the compartment external surface reacted via an aldehyde-amino reaction with glass surfaces chemically modified with amino groups. The nanoreactors preserved their architecture and activity after immobilization on the glass surface, and represent active biosensing surfaces for selective detection of sugar alcohols, with high sensitivity.

Adaptive Polymersome Nanoreactors

“…Penicillin acylase from E. coli was encapsulated inside the polymersomes to create nanoreactors which could convert 7‐aminodesacetoxycephalosporanic acid (7‐ADCA) and phenylglycine methyl ester (PGME) into cephalexin (semisynthetic cephalosporin antibiotic), capable of inhibiting bacterial growth. Additionally, they demonstrated the immobilization of polymersome nanoreactors on a solid support to generate “active surfaces”, which were utilized as efficient biosensors for sugar alcohols (Figure b) . E. coli glycerol facilitator (GlpF) was selected to transport sugar alcohols through the polymer membranes.…”

“…Additionally, they demonstrated the immobilization of polymersome nanoreactors on a solid support to generate "active surfaces", which were utilized as efficient biosensors for sugar alcohols (Figure 2b). [15] E. coli glycerol facilitator (GlpF) was selected to transport sugar alcohols through the polymer membranes. Ribitol dehydrogenase (RDH), which is able to catalyze a variety of sugar alcohol reactions, was encapsulated in PDMS-b-PMOXA polymersomes.…”

Nanostructured Surfaces through Immobilization of Self‐Assembled Polymer Architectures Using Thiol–Ene Chemistry

“…Moreover, their applicability as drug delivery platforms and reaction compartments has been shown in numerous studies . Stimuli‐responsive nanostructured surfaces become accessible through immobilization of adaptive or biohybrid self‐assembled architectures . The formation of polymeric nanoarchitectures via self‐assembly has been realized through different techniques, and few studies present routes to anchor polymer vesicles or polymer micelles on solid support.…”

“…A stronger interaction can be achieved using biotin‐streptavidin and host–guest interactions, however, this typically requires postmodification of the polymers or the assemblies as an additional reaction step. Direct covalent bonding has been achieved through imines and copper‐mediated click chemistry . Yet, a method that is applicable to different types of polymer nanoarchitectures and employs reaction conditions that can be tolerated by sensitive biomolecules will allow to proceed toward more advanced functional surface structures that possess additional biological activity.…”

'Active Surfaces' as Possible Functional Systems in Detection and Chemical (Bio) Reactivity

“…We were successful to introduce polymer compartments with selective permeability by inserting specific membrane proteins allowing a controlled diffusion through the synthetic membrane, and immobilized them on solid support to engineer functional surfaces with the desired functionality. [34] In this respect, we immobilized protein-polymer nanoreactors for sensitive detection of sugar alcohols by simultaneously encapsulating speinterface, films were oriented with the PEG domain in the water subphase and the PDMAEMA domain facing toward air. Laccase was successfully immobilized on the membrane either via immersion of solid-supported polymer films in enzyme solutions, or transfer of mixed ABC-enzyme films on silica slides (Fig.…”