Microgels formed by enzyme-mediated polymerization in reverse micelles with tunable activity and high stability

Bao, Song; Wu, Dong; Su, Teng; Wu, Qing; Wang, Qigang

doi:10.1039/c5ra02162f

Cited by 7 publications

(3 citation statements)

References 36 publications

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“…Wang and co-workers immobilized HRP in a self-catalyzed poly(N,N-dimethylacrylamide) gel in a water-in-oil (O/W) emulsion. 135 The oil phase was prepared using the surfactants tween 20 and span 80 in octane and acetylacetone. An aqueous solution of HRP, N,N-dimethylacrylamide 6 and 7 were added slowly to the oil phase to form the water-in-oil emulsion.…”

Section: Synthetic Polymersmentioning

confidence: 99%

Enzyme entrapment, biocatalyst immobilization without covalent attachment

2021

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Section: Synthetic Polymersmentioning

confidence: 99%

Enzyme entrapment, biocatalyst immobilization without covalent attachment

2021

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“…Polymer particles are often processed by coating their surface, mixing different monomers to prepare composite materials, and/or enclosing the beneficial materials (i.e., catalysts and bioagents) inside, depending on the intended applications. In general, polymer particles can be prepared by several methods including: (i) inverse emulsion polymerization, − (ii) Pickering emulsion polymerization, , (iii) solvent evaporation polymerization, , and (iv) mechanical stirring in a multi-phase system . A common feature of the above methods is the requirement of a surfactant/detergent substance to stabilize the multi-phase interface, where the preparation conditions must be optimized for each polymer material; however, to maintain both the stability of the dispersed phase and a good reactivity at the interface, the applied surfactant/detergent must remain in the obtained polymer particles.…”

Section: Introductionmentioning

confidence: 99%

Micro Sponge Balls: Preparation and Characterization of Sponge-like Cryogel Particles of Poly(2-hydroxyethyl methacrylate) via the Inverse Leidenfrost Effect

Takase

Shiomori

Okamoto

et al. 2022

ACS Appl. Polym. Mater.

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Here, we report a method to prepare cryogel particles with sponge-like mechanical properties, including high porosity and high elasticity. The preparation process of the cryogel particles of poly(2-hydroxythyl methacrylate) can be summarized in the two following steps: preparation of frozen droplets using the inverse Leidenfrost effect, followed by cryo-gelation by frozen polymerization. First, a polymer precursor was dropwise added into bulk liquid nitrogen (−196 °C). Then, frozen droplets were created by the inverse Leidenfrost effect, which were subsequently polymerized in liquid paraffine (−15 °C). After thawing and drying, the cryogel particles were obtained. The monolithic super-macroporous structure was observed by scanning electron microscopy (SEM). The mechanical properties of the cryogel particles were studied via compression−swelling tests. At maximum compression, the particles achieved 94.3% degree of deformation; remarkably, they returned to their original shape under the swelling state. The strategy proposed herein, which combines the inverse Leidenfrost effect with a cryopolymerization technique, could be applied to prepare various polymer particles without employing surfactants.

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“…In respect to enzyme immobilization in microgels, the classes of oxidoreductases and hydrolases have predominantly been reported. ,− An interesting approach for covalent immobilization of P450 BM3 in microgels was reported through enzymatic ligation by employing transpeptidase sortase A (Sa-SrtA) from Staphylococcus aureus . In the case of noncovalent immobilization, P450 BM3 was first immobilized to (PNIPAAm) microgels with N -vinylimidazole (Vim) as comonomer through electrostatic interactions .…”

Section: Introductionmentioning

confidence: 99%

MicroGelzymes: pH-Independent Immobilization of Cytochrome P450 BM3 in Microgels

et al. 2020

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Microgels are an emerging class of “ideal” enzyme carriers because of their chemical and process stability, biocompatibility, and high enzyme loading capability. In this work, we synthesized a new type of permanently positively charged poly(N-vinylcaprolactam) (PVCL) microgel with 1-vinyl-3-methylimidazolium (quaternization of nitrogen by methylation of N-vinylimidazole moieties) as a comonomer (PVCL/VimQ) through precipitation polymerization. The PVCL/VimQ microgels were characterized with respect to their size, charge, swelling degree, and temperature responsiveness in aqueous solutions. P450 monooxygenases are usually challenging to immobilize, and often, high activity losses occur after the immobilization (in the case of P450 BM3 from Bacillus megaterium up to 100% loss of activity). The electrostatic immobilization of P450 BM3 in permanently positively charged PVCL/VimQ microgels was achieved without the loss of catalytic activity at the pH optimum of P450 BM3 (pH 8; ∼9.4 nmol 7-hydroxy-3-carboxy coumarin ethyl ester/min for free and immobilized P450 BM3); the resulting P450-microgel systems were termed P450 MicroGelzymes (P450 μ-Gelzymes). In addition, P450 μ-Gelzymes offer the possibility of reversible ionic strength-triggered release and re-entrapment of the biocatalyst in processes (e.g., for catalyst reuse). Finally, a characterization of the potential of P450 μ-Gelzymes to provide resistance against cosolvents (acetonitrile, dimethyl sulfoxide, and 2-propanol) was performed to evaluate the biocatalytic application potential.

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Microgels formed by enzyme-mediated polymerization in reverse micelles with tunable activity and high stability

Abstract: This communication describes the preparation of microgels via enzyme-triggered inverse emulsion polymerization, which provides an effective method for immobilizing enzymes with tunable catalytic performance and high stability.

Cited by 7 publications

References 36 publications

Enzyme entrapment, biocatalyst immobilization without covalent attachment

Enzyme entrapment, biocatalyst immobilization without covalent attachment

Micro Sponge Balls: Preparation and Characterization of Sponge-like Cryogel Particles of Poly(2-hydroxyethyl methacrylate) via the Inverse Leidenfrost Effect

MicroGelzymes: pH-Independent Immobilization of Cytochrome P450 BM3 in Microgels

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