title: Engineering Systems with Spatially Separated Enzymes via Dual-Stimuli-Sensitive Properties of Microgels.
abstract: 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.
authors.affiliation: Department of Chemistry, M.V. Lomonosov Moscow State University , 119991 Moscow, Russia.
authors.forename: Larisa V
authors.initials: LV
authors.lastname: Sigolaeva
authors.affiliation: Institute of Physical Chemistry II, RWTH Aachen University , 52056 Aachen, Germany.
authors.forename: Olga
authors.initials: O
authors.lastname: Mergel
authors.affiliation: Department of Chemistry, M.V. Lomonosov Moscow State University , 119991 Moscow, Russia.
authors.forename: Evgeniy G
authors.initials: EG
authors.lastname: Evtushenko
authors.affiliation: Department of Chemistry, M.V. Lomonosov Moscow State University , 119991 Moscow, Russia.
authors.forename: Snezhana Yu
authors.initials: SY
authors.lastname: Gladyr
authors.affiliation: Institute of Physical Chemistry II, RWTH Aachen University , 52056 Aachen, Germany.
authors.forename: Arjan P H
authors.initials: AP
authors.lastname: Gelissen
authors.affiliation: Department of Chemistry, M.V. Lomonosov Moscow State University , 119991 Moscow, Russia.
authors.forename: Dmitry V
authors.initials: DV
authors.lastname: Pergushov
authors.affiliation: Department of Chemistry, M.V. Lomonosov Moscow State University , 119991 Moscow, Russia.
authors.forename: Ilya N
authors.initials: IN
authors.lastname: Kurochkin
authors.affiliation: Institute of Physical Chemistry II, RWTH Aachen University , 52056 Aachen, Germany.
authors.forename: Felix A
authors.initials: FA
authors.lastname: Plamper
authors.affiliation: Institute of Physical Chemistry II, RWTH Aachen University , 52056 Aachen, Germany.
authors.forename: Walter
authors.initials: W
authors.lastname: Richtering
mesh_terms: D000178:Acrylamides / Q000737:chemistry; D000180:Acrylic Resins / Q000737:chemistry; D000327:Adsorption; D000818:Animals; D016466:CHO Cells; D003412:Cricetulus; D011108:Polymers / Q000737:chemistry*; D013499:Surface Properties
chemical_list: D000178:Acrylamides; D000180:Acrylic Resins; D011108:Polymers; C052970:poly-N-isopropylacrylamide; C067295:N-isopropylacrylamide