Generation of New Enzymes via Covalent Modification of Existing Proteins

Diels–Alder Ligation of Peptides and Proteins

“…sequential combination of two or more bioconjugation steps which permits greater control over the conjugation process by using of heterobifunctional cross-linkers; [13] introduction of an unique cysteine into a particular position of the protein structure by site-directed mutagenesis, allowing the selective modification of this residue via thiolreactive reagents; [14] moderate selective N-terminus derivatization by carrying out the amine-acylation reactions under slightly acidic conditions. [15] The combination of biosynthetic methods for protein production and subsequent bioconjugation can not always provide the tools for all kinds of protein derivatization.…”

Protein scaffold of a designed metalloenzyme enhances the chemoselectivity in sulfoxidation of thioanisole

“…Understanding how proteins confer such reactivity and selectivity is important not only to providing deeper insight in biological functions, but also to its application in chemical transformations. [1][2][3][4][5][6][7][8][9][10][11] Toward this goal, much work has focused on the study of native metalloenzymes, such as cytochrome P-450s, a metalloenzyme with high chemoselectivity in the oxidation of C-H bonds. [12][13][14] These studies indicate that the protein scaffold is capable of creating the proper environment to modulate the reactive pathways of active intermediates so as to inhibit side reactions such as over oxidization.…”

Tailoring the Active Site of Chemzymes by Using a Chemogenetic‐Optimization Procedure: Towards Substrate‐Specific Artificial Hydrogenases Based on the Biotin–Avidin Technology

“…[3][4][5][6][7][8][9][10][11][12][13] With the hope of alleviating some of the inherent limitations of both enzymatic and organometallic catalysis, two approaches have recently witnessed a revival: 1) organocatalysis [14][15][16][17][18][19] and 2) artificial metalloenzymes based on either covalent [20,21] or supramolecular anchoring [22] of a catalytic moiety in a macromolecular host. [23][24][25][26][27][28][29][30] Inspired by the early works of Whitesides and Wilson, [22] we recently reported artificial metalloenzymes based on the biotin-avidin technology. [31][32][33][34][35] Herein, we report our efforts to produce substrate-specific and S-selective artificial metalloenzymes based on the biotin-avidin technology for the hydrogenation of a-acetamidodehydroamino acids.…”