Cytochrome P450-catalyzed insertion of carbenoids into N–H bonds

Wang, Z. Jane; Peck, Nicole E.; Renata, Hans; Arnold, Frances H.

doi:10.1039/c3sc52535j

Cited by 184 publications

(206 citation statements)

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“…Engineered heme proteins such as cytochrome P450 enzymes, myoglobin, and cyctochrome C have emerged as excellent catalysts for new‐to‐nature reactions,1, 2 including carbene‐transfer reactions such as cyclopropanations,3, 4, 5, 6, 7 olefinations, and8, 9 N−H,10, 11 Si−H,12 and B−H insertion reactions 13. These enzymes have proven amenable to optimization by both genetic methods and co‐factor replacement 14, 15, 16, 17, 18, 19, 20.…”

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confidence: 99%

An Artificial Heme Enzyme for Cyclopropanation Reactions

et al. 2018

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An artificial heme enzyme was created through self‐assembly from hemin and the lactococcal multidrug resistance regulator (LmrR). The crystal structure shows the heme bound inside the hydrophobic pore of the protein, where it appears inaccessible for substrates. However, good catalytic activity and moderate enantioselectivity was observed in an abiological cyclopropanation reaction. We propose that the dynamic nature of the structure of the LmrR protein is key to the observed activity. This was supported by molecular dynamics simulations, which showed transient formation of opened conformations that allow the binding of substrates and the formation of pre‐catalytic structures.

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confidence: 99%

An Artificial Heme Enzyme for Cyclopropanation Reactions

et al. 2018

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“…Frances Arnold's group at Caltech (Pasadena, CA) recently demonstrated that altered cofactor sites in enzymes can catalyze new reactions, notably the cyclopropanation of alkene via carbenes on the heme-containing cytochrome P450-BM3 scaffold from B. megaterium, never before observed in biocatalysis (15). Mutation of the conserved axial cysteine to serine enabled further novel catalysis, C-H amination (88) and N-H insertion (89). Lastly, a cytochrome P450 oxidase was discovered that catalyzed nitration of tryptophan derivatives (90).…”

Section: Wwwannualreviewsorg • Biocatalysis: a Status Reportmentioning

confidence: 96%

“…Cytochrome P450-BM3 variants were used to create secondary amines from a primary amine and a carbenoid, more specifically ethyldiazoacetate (89). The reaction is chemoselective: no double insertion to a tertiary amine was observed (Figure 11c) …”

Section: Wwwannualreviewsorg • Biocatalysis: a Status Reportmentioning

confidence: 99%

Biocatalysis: A Status Report

Bommarius

2015

Annu. Rev. Chem. Biomol. Eng.

139

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This review describes the status of the fields of biocatalysts and enzymes, as well as existing drawbacks, and recent advances in the areas deemed to represent drawbacks. Although biocatalysts are often highly active and extremely selective, there are still drawbacks associated with biocatalysis as a generally applicable technique: the lack of designability of biocatalysts; their limits of stability; and the insufficient number of well-characterized, ready-to-use biocatalysts. There has been significant progress on the following fronts: (a) novel protein engineering tools, both experimental and computational, have significantly enhanced the toolbox for biocatalyst development. (b) The deactivation of biocatalysts under various stresses can be described quantitatively via rational models. There are several cases of spectacular leaps of stabilization after accumulating all stabilizing mutations found in earlier rounds. The concept that stabilization against one type of stress commonly also stabilizes against other types of stress is now experimentally considerably better founded than a few years ago.

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“…As mall library of structurally distinct P450 BM3 mutants allowed the selective cyclopropanation of ad iverse set of styrenes.T his library was also effective for the insertion of carbenes into N À Hb onds. [17] Preliminary mechanistic studies suggest that Fe II is the resting state of the heme cofactor. This feature presents achallenge when attempting to reduce heme from Fe III to Fe II under physiological conditions,a sc ytochrome P450 enzymes have an elaborate gating mechanism to prevent electron transfer in the absence of substrate.Given the low affinity of the substrates for the proteins active site,t he desired conformational change is not possible.…”

Section: Cytochrome P450smentioning

confidence: 99%

Genetic Optimization of Metalloenzymes: Enhancing Enzymes for Non‐Natural Reactions

2016

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Artificial metalloenzymes have received increasing attention over the last decade as a possible solution to unaddressed challenges in synthetic organic chemistry. Whereas traditional transition-metal catalysts typically only take advantage of the first coordination sphere to control reactivity and selectivity, artificial metalloenzymes can modulate both the first and second coordination spheres. This difference can manifest itself in reactivity profiles that can be truly unique to artificial metalloenzymes. This Review summarizes attempts to modulate the second coordination sphere of artificial metalloenzymes by using genetic modifications of the protein sequence. In doing so, successful attempts and creative solutions to address the challenges encountered are highlighted.

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Cytochrome P450-catalyzed insertion of carbenoids into N–H bonds

Cited by 184 publications

References 40 publications

An Artificial Heme Enzyme for Cyclopropanation Reactions

An Artificial Heme Enzyme for Cyclopropanation Reactions

Biocatalysis: A Status Report

Genetic Optimization of Metalloenzymes: Enhancing Enzymes for Non‐Natural Reactions

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