6GMI

Genetic Engineering of an Artificial Metalloenzyme for Transfer Hydrogenation of a Self-Immolative Substrate in E. coli's Periplasm.

Summary for 6GMI

• Entry DOI: 10.2210/pdb6gmi/pdb
• Descriptor: Streptavidin, {N-(4-{[2-(amino-kappaN)ethyl]sulfamoyl-kappaN}phenyl)-5-[(3aS,4S,6aR)-2-oxohexahydro-1H-thieno[3,4-d]imidazol-4-yl]pentanamide}(chloro)[(1,2,3,4,5-eta)-1,2,3,4,5-pentamethylcyclopentadienyl]iridium(III), IRIDIUM (III) ION, ... (4 entities in total)
• Functional Keywords: biotin-binding protein, artificial transfer hydrogenase, beta barrel, streptavidin
• Biological source: Streptomyces avidinii
• Total number of polymer chains: 1
• Total formula weight: 20200.62

Authors

Rebelein, J.G. (deposition date: 2018-05-26, release date: 2018-10-10, Last modification date: 2024-01-17)

Primary citation

Zhao, J.,Rebelein, J.G.,Mallin, H.,Trindler, C.,Pellizzoni, M.M.,Ward, T.R.
Genetic Engineering of an Artificial Metalloenzyme for Transfer Hydrogenation of a Self-Immolative Substrate in Escherichia coli's Periplasm.
J. Am. Chem. Soc., 140:13171-13175, 2018

PubMed Abstract: 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.

PubMed: 30272972
DOI: 10.1021/jacs.8b07189

Experimental method

X-RAY DIFFRACTION (1.6 Å)