Activated formylglycine-generating enzymes and methods of producing and using the same
AssigneeR.P. Scherer Technologies, LLC
InventorsDavid Rabuka, Gregory W. deHart, Patrick Holder, Jeanne Baker
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Abstract
The present disclosure provides activated formylglycine-generating enzymes (FGE), methods of producing activated FGE, and their use in methods of producing a protein comprising a formylglycine (FGly) residue. The methods of producing activated FGE, as well as methods of use of activated FGE in producing FGly-containing proteins, include both cell-based and cell-free methods. Compositions and kits that find use, e.g., in practicing the methods of the present disclosure are also provided.
Claims16
A method of producing a protein comprising a formylglycine residue, the method comprising:
- expressing a combining an activated formylglycine-generating enzyme (FGE) and a protein comprising an FGE recognition site in a cell-free reaction mixture comprising Cu2+ to activate the FGE, under conditions in which the activated FGE converts a cysteine residue or a serine residue of the FGE recognition site to a formylglycine residue, to produce a protein comprising a formylglycine residue.
Dependencies are inferred from the claim text.