Engineering Systems with Spatially Separated Enzymes via Dual-Stimuli-Sensitive Properties of Microgels

This work examines the adsorption regime and the properties of microgel/enzyme thin films deposited onto conductive graphite-based substrates. The films were formed via two-step sequential adsorption. A temperature- and pH-sensitive poly(N-isopropylacrylamide)-co-(3-(N,N-dimethylamino)propylmethacry...

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Publié dans:Langmuir : the ACS journal of surfaces and colloids. - 1985. - 31(2015), 47 vom: 01. Dez., Seite 13029-39
Auteur principal: Sigolaeva, Larisa V (Auteur)
Autres auteurs: Mergel, Olga, Evtushenko, Evgeniy G, Gladyr, Snezhana Yu, Gelissen, Arjan P H, Pergushov, Dmitry V, Kurochkin, Ilya N, Plamper, Felix A, Richtering, Walter
Format: Article en ligne
Langue:English
Publié: 2015
Accès à la collection:Langmuir : the ACS journal of surfaces and colloids
Sujets:Journal Article Research Support, Non-U.S. Gov't Acrylamides Acrylic Resins Polymers poly-N-isopropylacrylamide 25189-55-3 N-isopropylacrylamide B7GFF17L9U
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520 |a This work examines the adsorption regime and the properties of microgel/enzyme thin films deposited onto conductive graphite-based substrates. The films were formed via two-step sequential adsorption. A temperature- and pH-sensitive poly(N-isopropylacrylamide)-co-(3-(N,N-dimethylamino)propylmethacrylamide) microgel (poly(NIPAM-co-DMAPMA microgel) was adsorbed first, followed by its interaction with the enzymes, choline oxidase (ChO), butyrylcholinesterase (BChE), or mixtures thereof. By temperature-induced stimulating both (i) poly(NIPAM-co-DMAPMA) microgel adsorption at T > VPTT followed by short washing and drying and then (ii) enzyme loading at T < VPTT, we can effectively control the amount of the microgel adsorbed on a hydrophobic interface as well as the amount and the spatial localization of the enzyme interacted with the microgel film. Depending on the biomolecule size, enzyme molecules can (in the case for ChO) or cannot (in the case for BChE) penetrate into the microgel interior and be localized inside/outside the microgel particles. Different spatial localization, however, does not affect the specific enzymatic responses of ChO or BChE and does not prevent cascade enzymatic reaction involving both BChE and ChO as well. This was shown by the methods of electrochemical impedance spectroscopy (EIS), atomic force microscopy (AFM), and amperometric analysis of enzymatic responses of immobilized enzymes. Thus, a novel simple and fast strategy for physical entrapment of biomolecules by the polymeric matrix was proposed, which can be used for engineering systems with spatially separated enzymes of different types 
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650 4 |a Research Support, Non-U.S. Gov't 
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650 7 |a Acrylic Resins  |2 NLM 
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700 1 |a Mergel, Olga  |e verfasserin  |4 aut 
700 1 |a Evtushenko, Evgeniy G  |e verfasserin  |4 aut 
700 1 |a Gladyr, Snezhana Yu  |e verfasserin  |4 aut 
700 1 |a Gelissen, Arjan P H  |e verfasserin  |4 aut 
700 1 |a Pergushov, Dmitry V  |e verfasserin  |4 aut 
700 1 |a Kurochkin, Ilya N  |e verfasserin  |4 aut 
700 1 |a Plamper, Felix A  |e verfasserin  |4 aut 
700 1 |a Richtering, Walter  |e verfasserin  |4 aut 
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