497-72-3Relevant academic research and scientific papers
Directing group-controlled regioselectivity in an enzymatic C-H bond oxygenation
Negretti, Solymar,Narayan, Alison R. H.,Chiou, Karoline C.,Kells, Petrea M.,Stachowski, Jessica L.,Hansen, Douglas A.,Podust, Larissa M.,Montgomery, John,Sherman, David H.
supporting information, p. 4901 - 4904 (2014/04/17)
Highly regioselective remote hydroxylation of a natural product scaffold is demonstrated by exploiting the anchoring mechanism of the biosynthetic P450 monooxygenase PikCD50N-RhFRED. Previous studies have revealed structural and biochemical evidence for the role of a salt bridge between the desosamine N,N-dimethylamino functionality of the natural substrate YC-17 and carboxylate residues within the active site of the enzyme, and selectivity in subsequent C-H bond functionalization. In the present study, a substrate-engineering approach was conducted that involves replacing desosamine with varied synthetic N,N-dimethylamino anchoring groups. We then determined their ability to mediate enzymatic total turnover numbers approaching or exceeding that of the natural sugar, while enabling ready introduction and removal of these amino anchoring groups from the substrate. The data establish that the size, stereochemistry, and rigidity of the anchoring group influence the regioselectivity of enzymatic hydroxylation. The natural anchoring group desosamine affords a 1:1 mixture of regioisomers, while synthetic anchors shift YC-17 analogue C-10/C-12 hydroxylation from 20:1 to 1:4. The work demonstrates the utility of substrate engineering as an orthogonal approach to protein engineering for modulation of regioselective C-H functionalization in biocatalysis.
SELECTIVE OXIDATION OF C-H BONDS OF MODIFIED SUBSTRATES BY P450 MONOOXYGENASE
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Page/Page column 14, (2012/07/31)
The present invention provides regio- and stereoselective oxidation of unactivated C—H bonds using an engineered mutant cytochrome P450 monooxygenase and an engineered substrate.
Total synthesis of methymycin
Oh, Hong-Se,Xuan, Richeng,Kang, Han-Young
experimental part, p. 4458 - 4463 (2009/12/25)
Methynolide and 10-epi-methynolide were synthesized from the necessary segments, which were prepared by the addition of Grignard reagents to the corresponding α-alkoxyketones utilizing 1,2-stereochemical selection based on Cram chelation control. Ring-clo
Engineering and analysis of a self-sufficient biosynthetic cytochrome P450 PikC fused to the RhFRED reductase domain
Li, Shengying,Podust, Larissa M.,Sherman, David H.
, p. 12940 - 12941 (2008/09/16)
Cytochrome P450 enzymes mediate important oxidative processes in biological systems including regio- and stereospecific hydroxylation and epoxidation reactions. The inherent requirement of these biomolecules for separate redox partner(s) significantly limits their application in biotechnology. To address this challenge, naturally occurring and/or bioengineered self-sufficient P450 systems with covalently fused redox partners have been utilized to harness their catalytic power. In this study, we describe the first in vitro characterization of a bacterial biosynthetic cytochrome P450 PikC fused to a heterologous reductase domain RhFRED that demonstrates single-component self-sufficiency. This novel fusion system not only produces a more active and effective biocatalyst but also suggests a general design for a universal reductase to generate diverse self-sufficient fusions for functional identification or industrial applications of biosynthetic P450s. Copyright
Macrolide biosynthesis: A single cytochrome P450, PicK, is responsible for the hydroxylations that generate methymycin, neomethymycin, and picromycin in Streptomyces venezuelae
Graziani, Edmund I.,Cane, David E.,Betlach, Melanie C.,Kealey, James T.,McDaniel, Robert
, p. 3117 - 3120 (2007/10/03)
The final step in the biosynthesis of methymycin, neomethymycin, and picromycin is an hydroxylation, shown to be carded out by the cytochrome P- 450 monooxygenase, PicK. Direct comparison of the relative k(cat)/K(m) values for the two substrates, YC-17 and narbomycin, showed a threefold rate preference of pick for narbomycin.
