Robust and Versatile Host Protein for the Design and Evaluation of Artificial Metal Centers

Johannes Fischer, Dominik Renn, Felix Quitterer, Anand Radhakrishnan, Meina Liu, Arwa Makki, Seema Arun Ghoprade, Magnus Rueping, Stefan T. Arold, Michael Groll, Jörg Eppinger

Research output: Contribution to journalArticlepeer-review

9 Scopus citations


Artificial metalloenzymes (ArMs) have high potential in biotechnological applications as they combine the versatility of transition-metal catalysis with the substrate selectivity of enzymes. An ideal host protein should allow high-yield recombinant expression, display thermal and solvent stability to withstand harsh reaction conditions, lack nonspecific metal-binding residues, and contain a suitable cavity to accommodate the artificial metal site. Moreover, to allow its rational functionalization, the host should provide an intrinsic reporter for metal binding and structural changes, which should be readily amendable to high-resolution structural characterization. Herein, we present the design, characterization, and de novo functionalization of a fluorescent ArM scaffold, named mTFP*, that achieves these characteristics. Fluorescence measurements allowed direct assessment of the scaffold's structural integrity. Protein X-ray structures and transition metal Förster resonance energy transfer (tmFRET) studies validated the engineered metal coordination sites and provided insights into metal binding dynamics at the atomic level. The implemented active metal centers resulted in ArMs with efficient Diels-Alderase and Friedel-Crafts alkylase activities.
Original languageEnglish (US)
Pages (from-to)11371-11380
Number of pages10
JournalACS Catalysis
Issue number12
StatePublished - Nov 12 2019


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