Biochemical and Structural Characterization of MycCI, a Versatile P450 Biocatalyst from the Mycinamicin Biosynthetic Pathway

Matthew D. DeMars, Fang Sheng, Sung Ryeol Park, Andrew N. Lowell, Larissa M. Podust, and David H. Sherman

ACS Chemical Biology,
2016, 11 (9), 2642–2654; DOI: 10.1021/acschembio.6b00479


Center members have built a team of researchers around the development and application of biocatalysts capable of selective C–H functionalization. This report from the Sherman and Podust groups discloses the characterization of MycCl, a versatile P450 enzyme. P450 monooxygenases (P450s) are some of nature’s most ubiquitous and versatile enzymes for performing oxidative metabolic transformations. Their unmatched ability to selectively functionalize inert C-H bonds has led to their increasing employment in academic and industrial settings for the production of fine and commodity chemicals.

In the present study, we investigated the reactivity of the P450 MycCI from the mycinamicin biosynthetic pathway toward a variety of macrocyclic compounds and discovered that the enzyme exhibits appreciable activity on several 16-membered ring macrolactones independent of their glycosylation state. These results were corroborated by performing equilibrium substrate binding experiments, steady-state kinetics studies, and x-ray crystallographic analysis of MycCI bound to its native substrate mycinamicin VIII. We also characterized TylHI, a homologous P450 from the tylosin pathway, and showed that its substrate scope is severely restricted compared to MycCI.

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