A5052685379- Cyclopropane Reaction Mechanisms
- Catalytic C–H Functionalization Methods
- Enzyme Catalysis and Immobilization
- Asymmetric Hydrogenation and Catalysis
- Synthesis and Catalytic Reactions
- CO2 Reduction Techniques and Catalysts
- Spectroscopy Techniques in Biomedical and Chemical Research
- Computational Drug Discovery Methods
- Click Chemistry and Applications
- Metalloenzymes and iron-sulfur proteins
- Advanced biosensing and bioanalysis techniques
- Photoacoustic and Ultrasonic Imaging
- Advanced Fluorescence Microscopy Techniques
- Image Processing Techniques and Applications
- Machine Learning in Materials Science
- Plant biochemistry and biosynthesis
- Carbon dioxide utilization in catalysis
- Protein Structure and Dynamics
- Electrocatalysts for Energy Conversion
- Innovative Microfluidic and Catalytic Techniques Innovation
- Molecular Sensors and Ion Detection
- Biotin and Related Studies
- Oxidative Organic Chemistry Reactions
- Catalysts for Methane Reforming
- Luminescence and Fluorescent Materials
University of Basel
2022-2025
ETH Zurich
2024
The University of Tokyo
2016-2024
Bunkyo University
2020
Tailored enzymes are crucial for the transition to a sustainable bioeconomy. However, enzyme engineering is laborious and failure-prone due its reliance on serendipity. The efficiency success rates of campaigns may be improved by applying machine learning map sequence-activity landscape based small experimental data sets. Yet, it often proves challenging reliably model large sequence spaces while keeping effort tractable. To address this challenge, we present an integrated pipeline combining...
Enantioselective C–H amidation offers attractive means to assemble C–N bonds synthesize high-added value, nitrogen-containing molecules. In recent decades, complementary enzymatic and homogeneous-catalytic strategies for have been reported. Herein, we report on an artificial metalloenzyme (ArM) resulting from anchoring a biotinylated Ir-complex within streptavidin (Sav). The ArM catalyzes the enantioselective of unactivated C(sp3)–H bonds. Chemogenetic optimization Ir cofactor Sav led...
Super-resolution vibrational microscopy is promising to increase the degree of multiplexing nanometer-scale biological imaging because narrower spectral linewidth molecular vibration compared fluorescence. However, current techniques super-resolution suffer from various limitations including need for cell fixation, high power loading, or complicated detection schemes. Here, we present reversible saturable optical Raman transitions (RESORT) microscopy, which overcomes these by using...
Design of experiments (DoE) plays an important role in optimizing the catalytic performance chemical reactions. The most commonly used DoE relies on response surface methodology (RSM) to model variable space experimental conditions with fewest number experiments. However, RSM leads exponential increase required as variables increases. Herein we describe a Bayesian optimization algorithm (BOA) optimize continuous parameters (e.g., temperature, reaction time, reactant and enzyme...
Carboxypeptidases (CPs) are a family of hydrolases that cleave one or more amino acids from the C-terminal peptides proteins. However, methodology to monitor activities CPs is poorly developed. Here, we present first versatile design strategy obtain activatable fluorescent probes for by utilizing intramolecular spirocyclization rhodamine translate "aliphatic carboxamide aliphatic carboxylate" structural conversion catalyzed into dynamic fluorescence activation. Based on this novel strategy,...
The selective functionalization of sp3 C–H bonds is a versatile tool for the diversification organic compounds. Combining attractive features homogeneous and enzymatic catalysts, artificial metalloenzymes offer an ideal means to selectively modify these inert motifs. Herein, we report on copper(I) heteroscorpionate complex embedded within streptavidin that catalyzes intramolecular insertion carbene into bonds. Target residues genetic optimization metalloenzyme were identified by quantum...
Iron-sulfur clusters have been reported to catalyze various redox transformations, including the multielectron reduction of CO2 hydrocarbons. Herein, we report design and assembly an artificial [Fe4S4]-containing Fischer-Tropschase relying on biotin-streptavidin technology. For this purpose, synthesized a bis-biotinylated [Fe4S4] cofactor with marked aqueous stability incorporated it in streptavidin. The effect second coordination sphere provided by protein environment was scrutinized cyclic...
Artificial metalloenzymes (ArMs) enable the integration of abiotic cofactors within a native protein scaffold, allowing for non-natural catalytic activities. Previous ArMs, however, have primarily relied on single cofactor systems, limiting them to only one function. Here we present an approach construct ArMs embedding two based biotin-streptavidin technology. By incorporating multiple into four binding sites streptavidin, engineered programmable tandem transformations including...
Artificial metalloenzymes (ArMs) enable the integration of abiotic cofactors within a native protein scaffold, allowing for non‐natural catalytic activities. Previous ArMs, however, have primarily relied on single cofactor systems, limiting them to only one function. Here we present an approach construct ArMs embedding two based biotin‐streptavidin technology. By incorporating multiple into four binding sites streptavidin, engineered programmable tandem transformations including...
Combinatorial use of an antibody-reporter enzyme conjugate and a fluorescence probe activated by the is powerful strategy for fluorescence-guided cancer surgery. However, conventional probes typical reporter enzymes are insufficiently bioorthogonal, resulting in high background signals non-target tissues. We screened library HMRef (rhodol derivative)-based bearing various sugar moieties, discovered that HMRef-β-D-fucose bioorthogonal mammalian systems, but metagenomic glycosidase, Td2F2....
Abstract Senescent cells promote cancer development and progression through chronic inflammation caused by a senescence‐associated secretory phenotype (SASP). Although various senotherapeutic strategies targeting senescent have been developed for the prevention treatment of cancers, technology in vivo detection evaluation cell accumulation has not yet established. Here, we identified activatable fluorescent probes dipeptidylpeptidase‐4 (DPP4) as an effective probe detecting enzymatic...
Spontaneously blinking fluorophores are powerful tools for live-cell super-resolution imaging under physiological conditions.
9-cyanopyronin is a promising scaffold that exploits resonance Raman enhancement to enable sensitive, highly multiplexed biological imaging. Here, we developed cyano-Hydrol Green (CN-HG) derivatives as scaffolds expand the color palette of 9-cyanopyronins. CN-HG exhibit sufficiently long wavelength absorption produce strong for near-infrared (NIR) excitation, and their nitrile peaks are shifted lower frequency than those The fluorescence strongly quenched due lack 10th atom, unlike pyronin...
Fluorogenic probes are essential tools for real-time visualization of dynamic intracellular processes in living cells, but so far, their design has been largely dependent on trial-and-error methods. Here we propose a quantum chemical calculation-based method rational prediction the fluorescence properties hydroxymethyl rhodamine (HMR)-based fluorogenic probes. Our computational analysis intramolecular spirocyclization reaction, which switches HMR derivatives, reveals that consideration...
Abstract Tailored enzymes hold great potential to accelerate the transition a sustainable bioeconomy. Yet, enzyme engineering remains challenging as it relies largely on serendipity and is, therefore, highly laborious prone failure. The efficiency success rates of campaigns may be improved substantially by applying machine learning construct comprehensive representation sequence-activity landscape from small sets experimental data. However, often proves reliably model large protein sequence...
Artificial (transfer) hydrogenases have been developed for organic synthesis, but they rely on precious metals. Native use Earth-abundant metals, these cannot be applied synthesis due, in part, to their substrate specificity. Herein, we report the design and development of manganese transfer based biotin-streptavidin technology. By incorporating bio-mimetic Mn(I) complexes into binding cavity streptavidin, through chemo-genetic optimization, obtained artificial enzymes that hydrogenate...
Abstract Single‐molecule enzyme activity assay is a platform that enables the analysis of activities at single proteoform level. The limitation targetable enzymes major drawback assay, but general reported to study single‐molecule esterases based on coupled using thioesters as substrate analogues. realized by developing highly water‐soluble thiol‐reacting probes phosphonate‐substituted boron dipyrromethene (BODIPY). system detection cholinesterase in blood samples level, and it shown...
Relying on ubiquitous alkenes, carboamination reactions enable the difunctionalization of double bond by concurrent formation a C–N and C–C single bond. In past years, several groups have reported elegant strategies for alkenes relying homogeneous catalysts or enzymes. Herein, we report an artificial metalloenzyme enantioselective dihydrofuran. Genetic optimization, combined with Bayesian optimization catalytic performance, afforded disubstituted tetrahydrofuran product in up to 22 TON 85%...
By anchoring a metal cofactor within host protein, so-called artificial metalloenzymes can be generated. Such hybrid catalysts combine the versatility of transition metals in catalyzing new-to-nature reactions with power genetic-engineering to evolve proteins. With aim gaining better control over second coordination-sphere interactions between streptavidin host-protein (Sav) and biotinylated cofactor, we engineered hydrophobic dimerization domain, borrowed from superoxide dismutase C (SOD),...
Abstract Relying on ubiquitous alkenes, carboamination reactions enable the difunctionalization of double bond by concurrent formation a C−N and C−C single bond. In past years, several groups have reported elegant strategies for alkenes relying homogeneous catalysts or enzymes. Herein, we report an artificial metalloenzyme enantioselective dihydrofuran. Genetic optimization, combined with Bayesian optimization catalytic performance, afforded disubstituted tetrahydrofuran product in up to 22...
Thanks to recent advances in enzyme repurposing, hemoproteins have gained significant attention as versatile biocatalysts that catalyze a variety of transformations, ranging from oxidation redox-neutral reactions. To complement these achievements, we report herein on our efforts repurpose myoglobin (Mb) into an asymmetric ketoreductase, using PhSiH3 reductant. Two rounds mutagenesis afforded double mutant capable reducing with high enantioselectivity broad range prochiral aliphatic and...
While natural terpenoid cyclases generate complex structures via cationic mechanisms, alternative radical cyclization pathways are underexplored. The metal-catalyzed hydrogen-atom-transfer (M-HAT) reaction offers an attractive means for hydrofunctionalizing olefins, providing access to terpenoid-like structures. Artificial metalloenzymes offer a promising strategy introducing M-HAT reactivity into protein scaffold. Herein, we report our efforts towards engineering artificial cyclase...