Cofactor displacing non-standard amino acid incorporation

Non-standard amino acid incorporation has proven to be a valuable tool in expanding the chemistry available to enzymes. These exciting new side chains open a plethora of new reaction modalities; however, said side chains tend to be sterically challenging. Most previous incorporation approaches have relied upon relatively porous, or cavity filled enzymes to accommodate said steric bulk. Our work has focused on expanding the set of available scaffolds for non-standard amino acid incorporation by turning to cofactor binding enzymes. Cofactors share many steric and hydrophobic properties with chemically interesting side chains. Filling the binding pocket for these cofactors with non-standard amino acids will therefore permit a host of new protein scaffolds to utilize novel reaction mechanisms.