Genetic Engineering of an Artificial Metalloenzyme for Transfer Hydrogenation 
of a Self-Immolative Substrate in Escherichia coil's Periplasm

Zhao, Jingming; Rebelein, Johannes G.; Mallin, Hendrik; Trindler, Christian; Pellizzoni, Michela M.; Ward, Thomas R.

doi:10.1021/jacs.8b07189

Datensatz

DATENSATZ AKTIONENEXPORT

Zur Ablage hinzufügen

Lokale TagsFreigabegeschichteDetailsÜbersicht

Freigegeben

Zeitschriftenartikel

Genetic Engineering of an Artificial Metalloenzyme for Transfer Hydrogenation of a Self-Immolative Substrate in Escherichia coil's Periplasm

MPG-Autoren

Es sind keine MPG-Autoren in der Publikation vorhanden

Externe Ressourcen

Es sind keine externen Ressourcen hinterlegt

Volltexte (beschränkter Zugriff)

Für Ihren IP-Bereich sind aktuell keine Volltexte freigegeben.

Volltexte (frei zugänglich)

Es sind keine frei zugänglichen Volltexte in PuRe verfügbar

Ergänzendes Material (frei zugänglich)

Es sind keine frei zugänglichen Ergänzenden Materialien verfügbar

Zitation

Zhao, J., Rebelein, J. G., Mallin, H., Trindler, C., Pellizzoni, M. M., & Ward, T. R. (2018). Genetic Engineering of an Artificial Metalloenzyme for Transfer Hydrogenation of a Self-Immolative Substrate in Escherichia coil's Periplasm. JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 140(41), 13171-13175. doi:10.1021/jacs.8b07189.

Zitierlink: https://hdl.handle.net/21.11116/0000-0009-EC82-A

Zusammenfassung

Artificial metalloenzymes (ArMs), which combine an abiotic metal cofactor with a protein scaffold, catalyze various synthetically useful transformations. To complement the natural enzymes' repertoire, effective optimization protocols to improve ArM's performance are required. Here we report on our efforts to optimize the activity of an artificial transfer hydrogenase (ATHase) using Escherichia coli whole cells. For this purpose, we rely on a self-immolative quinolinium substrate which, upon reduction, releases fluorescent umbelliferone, thus allowing efficient screening. Introduction of a loop in the immediate proximity of the Ir-cofactor afforded an ArM with up to 5-fold increase in transfer hydrogenation activity compared to the wild-type ATHase using purified mutants.