title: An artificial metalloenzyme with the kinetics of native enzymes.
abstract: Natural enzymes contain highly evolved active sites that lead to fast rates and high selectivities. Although artificial metalloenzymes have been developed that catalyze abiological transformations with high stereoselectivity, the activities of these artificial enzymes are much lower than those of natural enzymes. Here, we report a reconstituted artificial metalloenzyme containing an iridium porphyrin that exhibits kinetic parameters similar to those of natural enzymes. In particular, variants of the P450 enzyme CYP119 containing iridium in place of iron catalyze insertions of carbenes into C-H bonds with up to 98% enantiomeric excess, 35,000 turnovers, and 2550 hours-1turnover frequency. This activity leads to intramolecular carbene insertions into unactivated C-H bonds and intermolecular carbene insertions into C-H bonds. These results lift the restrictions on merging chemical catalysis and biocatalysis to create highly active, productive, and selective metalloenzymes for abiological reactions.
authors.affiliation: Department of Chemistry, University of California, Berkeley, CA 94720, USA.|Chemical Sciences Division, Lawrence Berkeley National Laboratory, 1 Cyclotron Road, Berkeley, CA 94720, USA.
authors.forename: P
authors.initials: P
authors.lastname: Dydio
authors.affiliation: Department of Chemistry, University of California, Berkeley, CA 94720, USA.|Chemical Sciences Division, Lawrence Berkeley National Laboratory, 1 Cyclotron Road, Berkeley, CA 94720, USA.
authors.forename: H M
authors.initials: HM
authors.lastname: Key
authors.affiliation: Department of Chemistry, University of California, Berkeley, CA 94720, USA.
authors.forename: A
authors.initials: A
authors.lastname: Nazarenko
authors.affiliation: Department of Chemistry, University of California, Berkeley, CA 94720, USA.
authors.forename: J Y-E
authors.initials: JY
authors.lastname: Rha
authors.affiliation: Department of Chemistry, University of California, Berkeley, CA 94720, USA.
authors.forename: V
authors.initials: V
authors.lastname: Seyedkazemi
authors.affiliation: Department of Chemical and Biomolecular Engineering, University of California, Berkeley, CA 94720, USA.|Molecular Biophysics and Integrated Bioimaging Division, Lawrence Berkeley National Laboratory, 1 Cyclotron Road, Berkeley, CA 94720, USA.
authors.forename: D S
authors.initials: DS
authors.lastname: Clark
authors.affiliation: Department of Chemistry, University of California, Berkeley, CA 94720, USA. jhartwig@berkeley.edu.|Chemical Sciences Division, Lawrence Berkeley National Laboratory, 1 Cyclotron Road, Berkeley, CA 94720, USA.
authors.forename: J F
authors.initials: JF
authors.lastname: Hartwig
mesh_terms: D055162:Biocatalysis; D000072496:Cytochrome P450 Family 19 / Q000737:chemistry* / Q000235:genetics; D007495:Iridium / Q000737:chemistry; D007700:Kinetics; D008667:Metalloproteins / Q000737:chemistry* / Q000235:genetics; D008697:Methane / Q000031:analogs & derivatives / Q000737:chemistry; D009154:Mutation; D011166:Porphyrins / Q000737:chemistry; D011487:Protein Conformation; D013237:Stereoisomerism
chemical_list: D008667:Metalloproteins; D011166:Porphyrins; C030011:carbene; D007495:Iridium; D000072496:Cytochrome P450 Family 19; D008697:Methane