2WHW

Selective oxidation of carbolide C-H bonds by engineered macrolide P450 monooxygenase

Summary for 2WHW

• Entry DOI: 10.2210/pdb2whw/pdb
• Related: 2BVJ 2C6H 2C7X 2CA0 2CD8 2VZ7 2VZM 2WI9
• Descriptor: CYTOCHROME P450 MONOOXYGENASE, PROTOPORPHYRIN IX CONTAINING FE, CYCLOTRIDECYL 3,4,6-TRIDEOXY-3-(DIMETHYLAMINO)-BETA-D-XYLO-HEXOPYRANOSIDE, ... (5 entities in total)
• Functional Keywords: antibiotic biosynthesis, cyp107l1, cytochrome p450, heme, iron, macrolide monooxygenase, metal-binding, monooxygenase, oxidoreductase, oxidoreductase antibiotic biosynthesis, pikc
• Biological source: STREPTOMYCES VENEZUELAE
• Total number of polymer chains: 2
• Total formula weight: 98499.49

Authors

Li, S.,Chaulagain, M.R.,Knauff, A.R.,Podust, L.M.,Montgomery, J.,Sherman, D.H. (deposition date: 2009-05-07, release date: 2009-10-27, Last modification date: 2023-12-13)

Primary citation

Li, S.,Chaulagain, M.R.,Knauff, A.R.,Podust, L.M.,Montgomery, J.,Sherman, D.H.
Selective Oxidation of Carbolide C-H Bonds by an Engineered Macrolide P450 Mono-Oxygenase.
Proc.Natl.Acad.Sci.USA, 106:18463-, 2009

PubMed Abstract: Regio- and stereoselective oxidation of an unactivated C-H bond remains a central challenge in organic chemistry. Considerable effort has been devoted to identifying transition metal complexes, biological catalysts, or simplified mimics, but limited success has been achieved. Cytochrome P450 mono-oxygenases are involved in diverse types of regio- and stereoselective oxidations, and represent a promising biocatalyst to address this challenge. The application of this class of enzymes is particularly significant if their substrate spectra can be broadened, selectivity controlled, and reactions catalyzed in the absence of expensive heterologous redox partners. In this study, we engineered a macrolide biosynthetic P450 mono-oxygenase PikC (PikC(D50N)-RhFRED) with remarkable substrate flexibility, significantly increased activity compared to wild-type enzyme, and self-sufficiency. By harnessing its unique desosamine-anchoring functionality via a heretofore under-explored "substrate engineering" strategy, we demonstrated the ability of PikC to hydroxylate a series of carbocyclic rings linked to the desosamine glycoside via an acetal linkage (referred to as "carbolides") in a regioselective manner. Complementary analysis of a number of high-resolution enzyme-substrate cocrystal structures provided significant insights into the function of the aminosugar-derived anchoring group for control of reaction site selectivity. Moreover, unexpected biological activity of a select number of these carbolide systems revealed their potential as a previously unrecorded class of antibiotics.

PubMed: 19833867
DOI: 10.1073/PNAS.0907203106

Experimental method

X-RAY DIFFRACTION (2.2 Å)