Engineering Orthogonal Methyltransferases to Create Alternative Bioalkylation Pathways

Herbert, Abigail J.; Shepherd, Sarah A.; Cronin, Victoria A.; Bennett, Matthew R.; Sung, Rehana; Micklefield, Jason

doi:10.1002/ange.202004963

Cited by 24 publications

(3 citation statements)

References 44 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…72, 79-81 For example, RnCOMT accepts carboxy-S-adenosyl-l-methionine (cxSAM) to enable carboxymethylation of 3,4-dihydroxybenzaldehyde with roughly 4:3 m-/p-selectivity. 82,83 The Y200L variant of this enzyme displayed improved m-/p-selectivity (64a' 64% and 65a' 3%), and similar selectivity trends were observed for 4-nitrocatechol. In a more recent study, an engineered halide methyltransferase (HMT) was evolved to accept ethyliodide as the alkyl donor to enable ethylation reactions.…”

Section: Alcohol Alkylationmentioning

confidence: 57%

Non-Native Site-Selective Enzyme Catalysis

Mondal

Snodgrass

Gomez

et al. 2023

Preprint

View full text Add to dashboard Cite

The ability to a site-selectively modify equivalent functional groups in a molecule has the potential to streamline syntheses and increase product yields by lowering step counts. Enzymes catalyze site-selective transformations throughout primary and secondary metabolism, but leveraging this capability for non-native substrates and reactions requires a detailed understanding of the potential and limitations of enzyme catalysis and how these bounds can be extended by protein engineering. In this review, we discuss representative examples of site-selective enzyme catalysis involving functional group manipulation and C-H bond functionalization. We include illustrative examples of native catalysis, but our focus is on cases involving non-native substrates and reactions often using engineered enzymes. We then discuss the use of these enzymes for chemoenzymatic transformations and target-oriented synthesis and conclude with a survey of tools and techniques that could expand the scope of non-native site-selective enzyme catalysis.

show abstract

Section: Alcohol Alkylationmentioning

confidence: 57%

Non-Native Site-Selective Enzyme Catalysis

Mondal

Snodgrass

Gomez

et al. 2023

Preprint

View full text Add to dashboard Cite

show abstract

“…In addition to the possibility of broadening the diversity of radical SAM MT products by transferring alternative alkyl chains, this might be interesting in terms of using SAM analogues for mechanistic studies as has been shown before [36,37,84,85] . Together with the increasing number of available enzyme structures, it might be possible to design enzyme variants with a broader or altered substrate range, as has been shown for MATs and conventional MTs [88–91] …”

Section: Resultsmentioning

confidence: 99%

Biomimetic S‐Adenosylmethionine Regeneration Starting from Multiple Byproducts Enables Biocatalytic Alkylation with Radical SAM Enzymes**

et al. 2023

View full text Add to dashboard Cite

S‐Adenosylmethionine (SAM) is an enzyme cofactor involved in methylation, aminopropyl transfer, and radical reactions. This versatility renders SAM‐dependent enzymes of great interest in biocatalysis. The usage of SAM analogues adds to this diversity. However, high cost and instability of the cofactor impedes the investigation and usage of these enzymes. While SAM regeneration protocols from the methyltransferase (MT) byproduct S‐adenosylhomocysteine are available, aminopropyl transferases and radical SAM enzymes are not covered. Here, we report a set of efficient one‐pot systems to supply or regenerate SAM and SAM analogues for all three enzyme classes. The systems’ flexibility is showcased by the transfer of an ethyl group with a cobalamin‐dependent radical SAM MT using S‐adenosylethionine as a cofactor. This shows the potential of SAM (analogue) supply and regeneration for the application of diverse chemistry, as well as for mechanistic studies using cofactor analogues.

show abstract

“…Enzymes are attractive alternatives for C-H functionalization because of their inherent chemoselectivity, sustainability, and potential to be optimized via protein engineering for tuning activity and stereoselectivity. [23][24][25][26][27][28] While natural enzymes capable of forging new carbon-carbon bonds via C(sp 3 )-H functionalization are rare and largely limited to specific substrates (e.g., methyl transfer reactions catalyzed by S-adenosylmethionine dependent enzymes) [29][30][31][32][33] , recent advances in protein engineering have expanded the repertoire of enzyme-catalyzed C-H functionalization via carbene transfer chemistry (Figure 2d-e). 23,34 Traditionally, C-H functionalization via metal-carbenoid insertion has been pursued through small molecule organometallic catalysts, including complexes with rhodium, [35][36][37] iridium, 38,39 and other metals.…”

Section: Introductionmentioning

confidence: 99%

Enantioselective Single and Dual a-C–H Bond Functionalization of Cyclic Amines via Enzymatic Carbene Transfer

Ren

Couture

Liu

et al. 2022

Preprint

View full text Add to dashboard Cite

Cyclic amines are ubiquitous structural motifs found in pharmaceuticals and biologically active natural products, making methods for their elaboration via direct C–H functionalization of considerable synthetic value. Herein, we report the development of an iron-based biocatalytic strategy for enantioselective a-C–H functionalization of pyrrolidines via a carbene transfer reaction with diazoacetone. Currently unreported for organometallic catalysts, this transformation can be accomplished in high yields, high catalytic activity and high stereoselectivity (up to 99:1 e.r. and 20,350 TON) using engineered variants of cytochrome P450 CYP119 from Sulfolobus solfataricus. This methodology was further extended to enable enantioselective a-C–H functionalization in the presence of ethyl diazoacetate as carbene donor (up to 89:11 e.r. and 8,920 TON), and the two strategies were combined to achieve a one-pot as well as a tandem dual C–H functionalization of the cyclic amine substrate with enzyme-controlled diastereo- and enantiodivergent selectivity. This biocatalytic approach is amenable to gram-scale synthesis and can be applied to drug scaffolds for late-stage C–H functionalization. This work provides an efficient and tunable method for direct asymmetric a-C–H functionalization of saturated N-heterocycles which should offer new opportunities for the synthesis, discovery, and optimization of bioactive molecules.

show abstract

Engineering Orthogonal Methyltransferases to Create Alternative Bioalkylation Pathways

Cited by 24 publications

References 44 publications

Non-Native Site-Selective Enzyme Catalysis

Non-Native Site-Selective Enzyme Catalysis

Biomimetic S‐Adenosylmethionine Regeneration Starting from Multiple Byproducts Enables Biocatalytic Alkylation with Radical SAM Enzymes**

Enantioselective Single and Dual a-C–H Bond Functionalization of Cyclic Amines via Enzymatic Carbene Transfer

Contact Info

Product

Resources

About