Immobilization of Water-Soluble HRP within Poly-<i>N</i>-isopropylacrylamide Microgel Particles for Use in Organic Media

Gawlitza, Kornelia; Georgieva, Radostina; Tavraz, Neslihan N.; Keller, Janos; Klitzing, Regine von

doi:10.1021/la403598s

Cited by 37 publications

(35 citation statements)

References 51 publications

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“…In this study, laccase could be activated by a factor of 5,000. HRP could be activated by some 3,000‐fold upon immobilization in a p‐NIPAM microgel in isopropanol (Gawlitza et al, ), which is close to the here achieved 5,000‐fold activation achieved in by solubilization with POx‐IDA in chloroform. CT could be activated by a factor of up to 1,000 when solubilized by AOT in organic solvents (Paradkar & Dordick, ), while the activation of this enzyme immobilized on nanoporous silica glass in hexane reached a 110‐fold activity (P. Wang, Dai, Waezsada, Tsao, & Davison, ).…”

Section: Discussionsupporting

confidence: 83%

“…However, they also work in organic solvents but are dramatically less active in their free form (Klibanov, ). The most common way to activate enzymes in organic media is to immobilize them on solid supports (Gawlitza, Georgieva, Tavraz, Keller, & von Klitzing, ; Mateo, Palomo, Fernandez‐Lorente, Guisan, & Fernandez‐Lafuente, ; Ruiz, Malavé, Felby, & Griebenow, ; Stepankova et al, ). This way the enzyme is accessible due to the large interface between the support and the solvent and at the same time stabilized.…”

Section: Introductionmentioning

confidence: 99%

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Poly(2‐oxazoline)s terminated with 2,2′‐imino diacetic acid form noncovalent polymer–enzyme conjugates that are highly active in organic solvents

Hijazi

Spiekermann

Krumm

et al. 2018

Biotech & Bioengineering

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A great limitation for the usability of free enzymes in organic solvents is their insolubility in these media. Some surfactants are capable of solubilizing enzymes in such media, but they are hard to remove. Covalent modification of enzymes with polymers has led to polymer-enzyme conjugates (PECs) that are soluble in organic solvents, but the process is quite elaborate. Poly(2-oxazoline)s (POx) with the end group 2,2′-imino diacetic acid were shown to form reversible, nano-sized noncovalent aggregates with enzymes. These PECs give clear solutions in organic solvents. The enzymes lysozyme, horseradish peroxidase (HRP), laccase, α-chymotrypsin (CT), catalase, and alcohol dehydrogenase could be solubilized in chloroform and toluene at concentrations of up to 2 mg protein/ml. Laccase, HRP, and CT were shown to survive the transfer into the organic medium and back to water in their active form. The distribution coefficient of the proteins between water and the organic solvent was shown to be dependent on the nature of the POx backbone. All three biocatalysts exhibit greatly enhanced activity in the respective organic solvent. K E Y W O R D Sbiocatalysis, enzyme, noncovalent polymer-enzyme conjugate, organic solvent, poly(2-oxazoline)

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Section: Discussionsupporting

confidence: 83%

Section: Introductionmentioning

confidence: 99%

Poly(2‐oxazoline)s terminated with 2,2′‐imino diacetic acid form noncovalent polymer–enzyme conjugates that are highly active in organic solvents

Hijazi

Spiekermann

Krumm

et al. 2018

Biotech & Bioengineering

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“…Gawlitza et al reported that the particles still exhibit a water content of 46 % in the interior after transferring the particle from water to isopropanol. Their interpretation is the formation of a water cage within the particles, which is formed during the solvent exchange [34]. Wang et al reported a similar behavior [21].…”

Section: Shift In the Particle Size Minimum From Bulk To Adsorbed Parmentioning

confidence: 94%

The impact of the cononsolvency effect on poly (N-isopropylacrylamide) based microgels at interfaces

2014

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The paper addresses the effect of solid interfaces on the cononsolvency effect for poly(N -iso propylacrylamide) based microgels containing different contents of the co-monomer allyl acetic acid (AAA). The cononsolvency effect is studied by dynamic light scattering (DLS) in solution and with atomic force microscopy (AFM) at surfaces against different mixtures of water and organic solvent (ethanol, iso-propanol, and tetrahydrofuran). For the studies at interfaces, the microgels are spin coated on silicon wafers that are precoated with poly(allylamine hydrochloride) (PAH). The minimum in particle volume due to cononsolvency shows a pronounced shift from 10-20 % of organic solvent to 40-50 % after deposition at the Si/PAH wafer. The strong shift indicates an increase of water to organic solvent ratio within the gel at the surface with respect to the bulk solution. In order to understand the increase of water to organic solvent ratio, shrinking/reswelling AFM experiments for different spincoating conditions and under ambient conditions are carried out. Spin coating from water instead from different solvent mixtures has no effect on the cononsolvency. In ambient conditions, the cononsolvency effect disappears

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“…Furthermore, microgel suspensions were studied as host media for proteins. In this approach, the network is swollen with the native buffer solution and protects the protein against denaturation and unfolding by a solvent [23,24,26].…”

Section: Introductionmentioning

confidence: 99%

Responsive Microgels at Surfaces and Interfaces

Wellert

Richter

Hellweg

et al. 2014

Zeitschrift Für Physikalische Chemie

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Stimuli responsive surface structures attract increasing attention due to a large variety of envisioned applications. The controlled organization of poly(N-isopropyl acrylamid), PNIPAM microgel particles at solid surfaces inspired numerous research activities. In this review article, we briefly discuss the swelling/deswelling properties of adsorbed microgel particles in comparison to the behavior in the bulk phase. The presence of the solid interface highly influences and changes their behavior with respect to the properties in solution. Furthermore, the confinement on a solid substrate allows the direct and in-situ investigation of the mechanical properties of the microgel particles. Additionally, we briefly review the research on microgel particles at liquid interfaces. At these interfaces new interesting effects occur. Moreover, we discuss some interesting work on potential applications. In this context, microgel particles are often used as an active component for responsive coatings of various functionality envisioning applications, e.g. in medicine, biotechnology, and nanooptics.Zusammenfassung: Responsive Oberflächenmaterialien finden aufgrund der damit verbundenen potentiellen technischen Anwendungsmöglichkeiten große Beachtung in der aktuellen Forschung. Die Möglichkeit, kolloidale Gelpartikel aus poly(N-Isopropylacrylamid), PNIPAM an festen Oberflächen kontrolliert anzuordnen, inspirierte vielfältige Forschungsakivitäten. In diesem Übersichtsartikel diskutieren wir das Quellverhalten adsorbierter Mikrogelpartikel im Vergleich zu

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Immobilization of Water-Soluble HRP within Poly-N-isopropylacrylamide Microgel Particles for Use in Organic Media

Cited by 37 publications

References 51 publications

Poly(2‐oxazoline)s terminated with 2,2′‐imino diacetic acid form noncovalent polymer–enzyme conjugates that are highly active in organic solvents

Poly(2‐oxazoline)s terminated with 2,2′‐imino diacetic acid form noncovalent polymer–enzyme conjugates that are highly active in organic solvents

The impact of the cononsolvency effect on poly (N-isopropylacrylamide) based microgels at interfaces

Responsive Microgels at Surfaces and Interfaces

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