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Periplasmic Screening for Artificial Metalloenzymes

Jeschek, Markus and Panke, Sven and Ward, Thomas R.. (2016) Periplasmic Screening for Artificial Metalloenzymes. Methods in enzymology, 580. pp. 539-556.

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Abstract

Artificial metalloenzymes represent an attractive means of combining state-of-the-art transition metal catalysis with the benefits of natural enzymes. Despite the tremendous recent progress in this field, current efforts towards the directed evolution of these hybrid biocatalysts mainly rely on the laborious, individual purification of protein variants rendering the throughput and hence the outcome of these campaigns feeble. We have recently developed a periplasmic screening platform for the directed evolution of an artificial metathase based on the streptavidin-biotin technology in the periplasm of the Gram-negative bacterium Escherichia coli . This periplasmic compartmentalization strategy comprises a number of compelling advantages, in particular with respect to artificial metalloenzymes, which lead to a drastic increase in the throughput of screening campaigns and additionally are of unique value for future in vivo applications. Therefore, we highlight here the benefits of this strategy and intend to propose a generalized guideline for the development of novel transition metal-based biocatalysts by directed evolution in order to extend the natural enzymatic repertoire.
Faculties and Departments:05 Faculty of Science > Departement Chemie
05 Faculty of Science > Departement Chemie > Chemie > Bioanorganische Chemie (Ward)
UniBasel Contributors:Ward, Thomas R. R.
Item Type:Article, refereed
Article Subtype:Research Article
Publisher:Elsevier
ISSN:0076-6879
Note:Publication type according to Uni Basel Research Database: Journal article
Language:English
Identification Number:
Last Modified:22 May 2019 08:30
Deposited On:22 May 2019 08:30

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