An Evolved Orthogonal Enzyme/Cofactor Pair

作者：Reynolds Evan W; McHenry Matthew W; Cannac Fabien; Gober Joshua G; Snow Christopher D; Brustad Eric M

来源：Journal of the American Chemical Society, 2016, 138(38): 12451-12458.

DOI：10.1021/jacs.6b05847

We introduce a strategy that expands the functionality of hemoproteins through orthogonal enzyme/heme pairs. By exploiting the ability of a natural heme transport protein, ChuA, to promiscuously import heme derivatives, we have evolved a cytochrome P450 (P450(BM3)) that selectively incorporates a nonproteinogenic cofactor, iron deuteroporphyrin IX (Fe-DPIX), even in the presence of endogenous heme. Crystal structures show that selectivity gains are due to mutations that introduce steric clash with the heme vinyl groups while providing a complementary binding surface for the smaller Fe-DPIX cofactor. Furthermore, the evolved orthogonal enzyme/cofactor pair is active in non-natural carbenoid-mediated olefin cyclopropanation. This methodology for the generation of orthogonal enzyme/cofactor pairs promises to expand cofactor diversity in artificial metalloenzymes.

• 出版日期2016-9-28