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Investigation of Horseradish Peroxidase Kinetics in an “Organelle-Like” Environment

Baumann, Patric and Spulber, Mariana and Fischer, Ozana and Car, Anja and Meier, Wolfgang. (2017) Investigation of Horseradish Peroxidase Kinetics in an “Organelle-Like” Environment. Small, 13 (17). p. 1603943.

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Official URL: http://edoc.unibas.ch/55590/

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Abstract

In order to mimic cell organelles, artificial nanoreactors have been investigated based on polymeric vesicles with reconstituted channel proteins (outer membrane protein F) and coencapsulated enzymes horseradish peroxidase (HRP) along with a crowding agent (Ficoll or polyethylene glycol) inside the cavity. Importantly, the presence of macromolecules has a strong impact on the enzyme kinetics, but no influence on the integrity of vesicles up to certain concentrations. This particular design allows for the first time the determination of HRP kinetics inside nanoreactors with crowded milieu. The values of the Michaelis Menten constant (K-m) measured for HRP in a confined space (encapsulated in nanoreactors) in the absence of macromolecules are approximate to 50% lower than in free conditions, and the presence of a crowding agent results in a further pronounced decrease. These results clearly suggest that activities of enzymes in confined spaces can be tuned by varying the concentrations of crowding compounds. The present investigation represents an advance in nanoreactor design by considering the influence of environmental factors on enzymatic performance, and it demonstrates that both encapsulation and the presence of a crowding environment increase the enzyme-substrate affinity.
Faculties and Departments:05 Faculty of Science > Departement Chemie
05 Faculty of Science > Departement Chemie > Chemie > Makromolekulare Chemie (Meier)
UniBasel Contributors:Meier, Wolfgang P.
Item Type:Article, refereed
Article Subtype:Research Article
Publisher:Wiley
ISSN:1613-6810
e-ISSN:1613-6829
Note:Publication type according to Uni Basel Research Database: Journal article
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Last Modified:10 Jul 2017 07:49
Deposited On:10 Jul 2017 07:49

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