Yong‐Zheng Chen

A three-component reaction of N -substituted hydrazines, aldehydes, and carboxylic acids has been achieved for the synthesis of N -acylhydrazones via the preferential formation of imines and the selective monoacylation of N -substituted hydrazones. The reaction has a wide substrate scope and functional group tolerance. The synthetic utility has been illustrated by the facile construction of valuable bioactive molecules and late-stage functionalization of pharmaceuticals.

Expanding enzyme catalytic diversity is essential to unlocking the full potential of biocatalysis for green chemistry. Halohydrin dehalogenases are promiscuous biocatalysts that typically mediate the intramolecular dehalogenative cyclization of haloalcohols to form cyclic ethers, as well as the ring-opening of these O-heterocycles with various anionic nucleophiles. Here we report that engineered variants of the halohydrin dehalogenase HheC instead catalyze an intermolecular dehalogenative hydroxylation of ε-haloalcohols with remote enantiocontrol. Mechanistic investigations establish that an Asp-Arg dyad serves as the key catalytic motif responsible for this unconventional dehalogenation pathway. Through directed evolution, we further improve both the remote stereoselectivity and catalytic efficiency of the enzyme and systematically profile its substrate scope. The biocatalytic process is also successfully scaled to preparative and gram scales for the synthesis of chiral ε-haloalcohols and ε-diols. Our work uncovers an unusual dehalogenation mechanism and a practical platform for enantiocontrolled synthesis of chiral ε-substituted alcohols. Halohydrin dehalogenases are promiscuous biocatalysts that typically mediate the intramolecular dehalogenative cyclization of haloalcohols to form cyclic ethers, as well as the ring-opening of these O-heterocycles with various anionic nucleophiles. Here, the authors report that engineered variants of the halohydrin dehalogenase HheC instead catalyze an intermolecular dehalogenative hydroxylation of εhaloalcohols with remote enantiocontrol.

PDLIM2, a PDZ-LIM domain-containing protein expressed highest in the lung and immune cells, serves as a unique tumor suppressor and immune modulator, mainly by turning off the activation of the master transcription factors NF-κB and STAT3. While its role in cancer is established, the involvement of PDLIM2 in viral infection remains unclear. Here we analyzed public gene expression data of blood leukocytes, bronchoalveolar lavage cells, and lung tissues from uninfected healthy humans and those infected with the respiratory virus severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) or influenza. We found that PDLIM2 expression was repressed by viral infection, and notably, this repression correlated with the severity of infectious diseases. Consistently, the expression level of PDLIM2 was negatively associated with NF-κB and STAT3 activity across a diverse range of cell types, such as macrophages, monocytes, neutrophils, T cells, alveolar type 1 and 2 epithelial cells, airway epithelial cells, and fibroblasts. Accordingly, cells with low PDLIM2 expression exhibited aberrant activation of signaling pathways essential for cellular functions and immune responses. These findings highlight PDLIM2 repression as a common mechanism underlying human viral infectious diseases and suggest PDLIM2 as a potential biomarker and therapeutic target for disease prognosis, prevention, and treatment.

-nucleophilic ring-opening reaction of spiro-vinylcyclopropyl oxindoles introducing an azido or a substituted amino group into the C3 position has been developed. Using palladium (0) as the catalyst, the racemic spiro-vinylcyclopropyl oxindoles can not only be converted to their chiral diastereoisomers via a stereoisomerization process but also generate a series of azido-substituted oxindole derivatives through the nucleophilic azide addition or be transformed to tertiary amine-substituted oxindoles through a simple ring-opening process. Notably, the azide products can serve as an important synthetic intermediate to successfully convert to other 3-substituted oxindole derivatives and the spiro[furan-3,3'-oxindoles], which exhibit good antitumor activity in vitro, thus demonstrating their potential applications in medicinal chemistry.

This study innovates pharmaceutical engineering education with an AI-driven CDMO framework, addressing traditional curricular gaps via targeted solutions. CDMO-oriented teaching cases enhance students' domain expertise, practical skills, and STEM competition competitiveness-evidenced by higher-tier awards and recognized pedagogical effectiveness. This work provides a replicable paradigm for nurturing interdisciplinary talents meeting modern pharmaceutical industry needs.

With the expansion of unmanned aerial vehicles (UAVs) into complex three-dimensional (3D) terrains for reconnaissance, rescue, and related missions, traditional path planning methods struggle to meet multi-constraint and multi-objective requirements. Existing swarm intelligence algorithms, limited by the "no free lunch" theorem, also face challenges when the standard Information Acquisition Optimizer (IAO) is applied to such tasks, including low exploration efficiency in high-dimensional search spaces, rapid loss of population diversity, and improper boundary handling. To address these issues, this study proposes a Multi-Strategy Enhanced Information Acquisition Optimizer (MEIAO). First, a Levy Flight-based information collection strategy is introduced to leverage its combination of short-range local searches and long-distance jumps, thereby broadening global exploration. Second, an adaptive differential evolution operator is designed to dynamically balance exploration and exploitation via a variable mutation factor, while crossover and greedy selection mechanisms help maintain population diversity. Third, a globally guided boundary handling strategy adjusts out-of-bound dimensions to feasible regions, preventing the generation of low-quality paths. Performance was evaluated on the CEC2017 (dim = 30/50/100) and CEC2022 (dim = 10/20) benchmark suites by comparing MEIAO with eight algorithms, including VPPSO and IAO. Based on the mean, standard deviation, Friedman mean rank, and Wilcoxon rank-sum tests, MEIAO demonstrated superior performance in local exploitation of unimodal functions, global exploration of multimodal functions, and complex adaptation on composite functions while exhibiting stronger robustness. Finally, MEIAO was applied to 3D mountainous UAV path planning, where a cost model considering path length, altitude standard deviation, and turning smoothness was established. The experimental results show that MEIAO achieved an average path cost of 253.9190, a 25.7% reduction compared to IAO (341.9324), with the lowest standard deviation (60.6960) among all algorithms. The generated paths were smoother, collision-free, and achieved faster convergence, offering an efficient and reliable solution for UAV operations in complex environments.

Enantioconvergent catalysis efficiently converts racemic substrates into single enantiomer products, overcoming the 50% yield limitation of kinetic resolutions while avoiding stereoisomer separation challenges. Despite advances in enantioconvergent chemo- and biocatalysis, expanding the substrate scope and enhancing catalytic versatility remain key challenges for the broader implementation of this transformative synthetic strategy. Herein, we present a unique biocatalytic platform for the enantioconvergent synthesis of vicinal diols using racemic epoxides or β-haloalcohols as starting materials. This methodology exploits engineered halohydrin dehalogenases to catalyze nitrite-mediated epoxide ring-opening with complementary regio- and enantioselectivity, enabling complete stereoconvergence through dual O-attack pathways. A variety of epoxides and β-haloalcohols are enantioconvergently converted into the corresponding enantioenriched vicinal diols on a preparative scale, achieving isolated yields of up to 89% and optical purities as high as a 96:4 enantiomeric ratio. Additionally, the enantioconvergent mechanism is elucidated through structure determination of the engineered enzyme, complemented by molecular dynamics and quantum mechanics/molecular mechanics calculations. Our study not only expands the enzymatic toolkit for creating chiral molecules but also opens up a methodology for enantioconvergent synthesis.

Expanding biocatalytic reactions to access previously inaccessible chemical transformations is of significant demand. Halohydrin dehalogenases represent a class of remarkably versatile biocatalysts that have been successfully explored in the asymmetric synthesis and transformation of epoxides as well as oxetanes. In this study, we extended their catalytic capabilities to establish a biocatalytic approach for the kinetic resolution of δ-haloalcohols. This approach enables the highly efficient and enantiocomplementary synthesis of diverse chiral δ-haloalcohols and tetrahydrofurans with good isolated yields (up to 50%) and high optical purities (up to >99% ee) on a preparative scale. These molecules are of significant interest in both synthetic and pharmaceutical chemistry. Remarkably, this biocatalytic method tolerates high substrate concentration up to 185 g/L, underscoring its synthetic potential for industrial applications. Furthermore, we propose a potential reaction mechanism based on a comprehensive analysis of various mutants and docking studies. Biocatalysis offers exceptional selectivity in asymmetric synthesis, yet the range of catalytic reactions remains limited. Here, the authors expand the capabilities of halohydrin dehalogenases to achieve the kinetic resolution of δ-haloalcohols, enabling efficient synthesis of chiral δ-haloalcohols and tetrahydrofurans with high optical purity.

This systematic analysis suggests setting a 2 mm and longer as clear surgical margin for TORS. Margins>2 mm are associated with superior postoperative PFS rate and prolonged PFS time in HPV-positive oropharyngeal carcinoma patients.

Axially chiral allenes serve as versatile building blocks in organic synthesis and are important motifs in natural products. While enantioselective syntheses of disubstituted allenes have been well-established, catalytic construction of trisubstituted and tetrasubstituted allenes with high enantiopurity remains a formidable challenge. In this study, we present a biocatalytic strategy for the stereodivergent synthesis of axially chiral tri- and tetrasubstituted allenols via engineered P450pyr monooxygenase-catalyzed desymmetric hydroxylation of C(sp 3 )–H bonds. Through structure-guided semirational mutagenesis, enantiocomplementary P450pyr variants were developed to deliver both ( R )- and ( S )-configured allenols in a highly regio-, chemo-, and enantioselective manner (up to 99% ee ). Molecular docking studies elucidated the structural basis for the enhanced and inverted stereocontrol in the engineered mutants, correlating active-site interactions with stereochemical outcomes. This enzymatic platform represents the first example of P450-catalyzed hydroxylation for constructing axially chiral tri- and tetrasubstituted allenes that are otherwise challenging to access.

antitumor activities of the highly valuable and modifiable synthetic products.

Background: PDLIM2, a PDZ-LIM domain-containing protein expressed highest in the lung and immune cells, serves as a unique tumor suppressor and immune modulator, mainly by turning off the activation of the master transcription factors NF-κB and STAT3. While its role in cancer is established, the involvement of PDLIM2 in viral infection remains unclear. Results: Here, we analyzed public gene expression data of blood leukocytes, bronchoalveolar lavage cells, and lung tissues from uninfected healthy humans and those infected with the respiratory virus SARS-CoV-2 or influenza. We found that PDLIM2 expression was repressed by viral infection, and notably, this repression correlated with the severity of infectious diseases. Consistently, the expression level of PDLIM2 was negatively associated with NF-κB and STAT3 activity across a diverse range of cell types, such as macrophages, monocytes, neutrophils, T cells, alveolar type 1 and 2 epithelial cells, airway epithelial cells, and fibroblasts. Accordingly, cells with low PDLIM2 expression exhibited aberrant activation of signaling pathways essential for cellular functions and immune responses. Conclusions: These findings highlight PDLIM2 repression as a common mechanism underlying human viral infectious diseases and suggest PDLIM2 as a potential biomarker and therapeutic target for disease prognosis, prevention, and treatment.

Axially chiral alkylidene cycloalkanes are important scaffolds found in various bioactive molecules, yet their synthetic methods remain underdeveloped. In particular, the construction of alkylidene cycloalkanes bearing both axial and central chiralities poses a significant challenge. In this work, we present an efficient and green strategy for synthesizing axially chiral alkylidene cyclopentanones and cyclopentanols through a remotely controlled kinetic resolution (KR) approach. Directed evolution of an alcohol dehydrogenase (TbSADH) from Thermoethanolicus brockii yielded a variant (TbSADH-I86A/W110R/Y267L) capable of catalyzing the stereoselective reduction, affording a diverse range of axially and centrally chiral alkylidene cyclopentanols with high enantio- and diastereoselectivity, as well as excellent selectivity factors (ee up to 99%, dr up to >99:1, and s up to >100). Additionally, we demonstrated the stereodivergent synthesis of alcohols featuring both axial and central chirality via chemoenzymatic methods. To elucidate the molecular recognition mechanisms, molecular docking studies were conducted, revealing that the altered active pocket geometry of the mutant enzyme enhances both activity and enantioselectivity. This methodology not only provides a novel biocatalytic platform for accessing nonatropisomeric axially chiral structures with dual chirality elements but also underscores the potential of enzymatic strategies as powerful tools for remote stereocontrol in synthetic organic chemistry.

Abstract Using visible light to drive NADH regeneration is an economically viable and environmentally sustainable technique. However, it necessitates a metal hydride (MH, [CpRh(bpy)(H 2 O)] 2+ ) as a synergist, and the high cost of the Rh noble metal significantly impedes the development and application of in‐situ NADH regeneration. Therefore, in this study, single‐atom Rh was immobilized onto the CdS QDs@SiO 2 combination via a consecutive ball‐milling technique in combination with ionic layer adsorption and substitution. Subsequently, an enhanced photo‐metal synergistic catalysis system for the recyclable regeneration of NADH was developed. In this composite, the single‐atom Rh serves two main functions: It acts as an electrical medium and a metal catalyst, which regulates the activity and selectivity of the regenerated NADH. This study has successfully addressed the key scientific issues regarding the low electron transport rate and the recycling of the Rh noble metal during catalysis. Results confirm that single‐atom Rh is successfully immobilized onto the CdS QDs@SiO 2 combination (Rh‐CdS@SiO 2 ) and exhibits a faster electron transport and enhanced selectivity. Under blue light (LED, 420 nm) irradiation, the Rh‐CdS@SiO 2 photo‐metal catalyst shows a 25‐fold increase in recyclable operability and achieves a 68% regeneration yield of NADH in just 4 min. Moreover, (S)‐(+)‐4‐phenyl‐2‐butanol can be obtained with the regenerated NADH as the coenzyme of P450 enzyme catalysis.

Abstract A simple photoinduced radical borylation, phosphonation, sulfurization and selenization of various aryl iodides to construct the structurally diverse C‐B/P/S/Se bonds has been developed. Using HCO 2 Na as the reductant, a wide range of aryl iodides including the substituted iodobenzenes, heteroaromatic iodides, aromatic iodides derived from the pharmaceutically active molecules could be utilized to react with the bis(pinacolato)diboron, triethyl phosphite, (RS) 2 or (RSe) 2 to obtain a variety of aryl borates, aryl phosphates, aryl sulfides and aryl selenides in general moderate to good yields.The advantage of this protocol is also highlighted by its no extra metal/photocatalyst, gram‐scale synthesis and the further functional transformations of the products.

Skeletal editing can rapidly forge valuable derivatives from parent molecule by late-stage simple transformations. In their Communication (e202501966), Wen-Yong Han et al. developed a Rh2(esp)2-catalyzed indole-quinoline transmutation using an α-diazotrifluoroethyl sulfonium salt, wherein the formal trifluoromethyl Rh-carbynoid (CF3C+ = Rh), a trifluoromethyl cationic carbyne (CF3C+:) equivalent, was involved.

Abstract Skeleton editing is an emerging and powerful tool in organic chemistry because it can simplify synthetic procedures towards complex molecules. Herein, we present an approach for indole‐quinoline transmutation through rhodium‐catalyzed single‐carbon insertion into the C2( sp 2 )─C3( sp 2 ) bond of an indole with an α‐diazotrifluoroethyl sulfonium salt that we developed. This protocol involves a formal trifluoromethyl rhodium carbynoid (CF 3 C + = Rh) resembling a trifluoromethyl cationic carbyne (CF 3 C + :), allowing facile access to an array of quinolines in moderate to high yields. Potential applications in the late‐stage skeletal editing of pharmaceutical and natural product derivatives, preparation of adapalene analogs, scaled‐up synthesis, and transformations of products are highlighted. Finally, a computational study was conducted to support the envisioned mechanism.

Abstract Skeleton editing is an emerging and powerful tool in organic chemistry because it can simplify synthetic procedures towards complex molecules. Herein, we present an approach for indole‐quinoline transmutation through rhodium‐catalyzed single‐carbon insertion into the C2( sp 2 )─C3( sp 2 ) bond of an indole with an α‐diazotrifluoroethyl sulfonium salt that we developed. This protocol involves a formal trifluoromethyl rhodium carbynoid (CF 3 C + = Rh) resembling a trifluoromethyl cationic carbyne (CF 3 C + :), allowing facile access to an array of quinolines in moderate to high yields. Potential applications in the late‐stage skeletal editing of pharmaceutical and natural product derivatives, preparation of adapalene analogs, scaled‐up synthesis, and transformations of products are highlighted. Finally, a computational study was conducted to support the envisioned mechanism.

Although biocatalysis offers complementary or alternative approaches to traditional synthetic methods, the limited range of available enzymatic reactions currently poses challenges in synthesizing a diverse array of desired compounds. Consequently, there is a significant demand for developing novel biocatalytic processes to enable reactions that were previously unattainable. Herein, we report the discovery and subsequent protein engineering of a unique halohydrin dehalogenase to develop a biocatalytic platform for enantioselective formation and ring-opening of oxetanes. This biocatalytic platform, exhibiting high efficiency, excellent enantioselectivity, and broad scopes, facilitates the preparative-scale synthesis of chiral oxetanes and a variety of chiral γ-substituted alcohols. Additionally, both the enantioselective oxetane formation and ring-opening processes are proven scalable for large-scale transformations at high substrate concentrations, and can be integrated efficiently in a one-pot, one-catalyst cascade system. This work expands the enzymatic toolbox for non-natural reactions and will promote further exploration of the catalytic repertoire of halohydrin dehalogenases in synthetic and pharmaceutical chemistry. Oxetane is a four-membered, oxygen-containing heterocyclic compound of importance in medicinal chemistry and drug development. Here, the authors report the discovery and subsequent protein engineering of a halohydrin dehalogenase, and develop a biocatalytic platform for enantioselective formation and ring-opening of oxetanes.

Abstract Chiral sulfoxides are essential in fields of pharmaceutical industry and asymmetric synthesis, requiring high enantioselectivity for their diverse applications. Biocatalytic methods, utilizing oxidases (e. g., BVMO, P450MO) and reductases (e. g., Msr, DMSO reductase), have proven to be efficient and sustainable alternatives to chemical synthesis. This review highlights progress made over the past decade in enzymatic approaches, including asymmetric oxidation, kinetic resolution, and cascade catalysis, with contributions from enzyme engineering to enhance activity, selectivity, and substrate scope, underscoring the potential of biocatalysis as a green and practical solution for producing enantiopure sulfoxides.

N-acyl/sulfonyl-α-phosphonated 1,2,3,4-tetrahydroiso-quinolines (THIQs) are significant structural motifs in organic synthesis and drug discovery. However, the one-pot approach enabling direct difunctionalization of THIQs remains challenging. Herein we report a photomediated one-pot multicomponent cascade reaction to access N-acyl/sulfonyl-α-phosphonated THIQs via twice acyl/sulfonyl iminium. This method features air as an oxidant, high atom and step economy, and mild conditions.

Enantiopure 3-hydroxyoxindoles are one class of basic functional molecules that hold particular interest in medicinal chemistry and drug discovery due to their diverse pharmacological properties. While many chemical methods have been developed for producing these molecules, there remains a continuous demand for more efficient and greener approaches. Herein, we present a novel dual-enzyme biocatalytic platform for the enantiocomplementary synthesis of chiral 3-hydroxy-3-hydroxymethyloxindoles, compounds that have not previously been synthesized stereoselectively. This biocatalytic platform involves the halohydrin dehalogenase-catalyzed kinetic resolution of racemic spiro-epoxyoxindoles with nitrite, paired with the epoxide hydrolase-catalyzed enantiospecific hydrolysis of the residual enantiopure spiro-epoxyoxindoles. Both enzymatic processes demonstrate high catalytic selectivity and efficiency, enabling the preparative synthesis of various ( R )- and ( S )-3-hydroxy-3-hydroxymethyloxindoles with high yields (up to 50%) and optical purities (up to >99% ee ). In addition, useful transformations of the chiral products were conducted to further showcase the scalability and applicability of the biocatalytic platform.

The work reports an improved method for the efficient, stereoselective and industrially feasible synthesis of the key intermediate for ( S )-indoxacarb, which features purification by filtration, using aqueous TBHP as an oxidant and mild conditions.

A photoinduced, tunable fluoroalkylation- or sulfonylation/cyclization of β -4′-methindolylstyrene to access 4-sulfonyl-, 4-fluoroalkyl- or 2-fluoroalkyl-tetrahydrobenzo[ cd ]-indoles via EDA complexes is described.

An artificial biocascade platform enables the enantiocomplementary synthesis of chiral 3-hydroxy-γ-butyrolactone from bio-based 1,3-dichloropropanol.

A convenient electrochemical oxidative cascade cyclization of alkenes equipped with pendant alcohols with general nucleophiles was developed. Using readily available diarylmethanimine and carboxylic acids as nucleophilic sources, a broad range of internal alkene and terminal alkene substrates could produce RCO 2 - and Ar 2 CN-functionalized O -heterocycles in moderate to high yields without the requirement for external oxidants and metals. These resulting products can subsequently be hydrolyzed to yield valuable NH 2 - and OH-functionalized tetrahydrofurans and tetrahydropyranes under mild conditions. Importantly, the efficient conversion of secondary alcohol products to amines with complete inversion of configuration enhances the methodology, enabling the construction of 2-aryl-3-amino tetrahydrofuran with high and complementary diastereoselectivity.

High Resolution Image Download MS PowerPoint Slide While enzymatic epoxidation of activated olefins by P450s has been well-established, chemo- and enantioselective epoxidation of unactivated olefins remains a formidable challenge, mainly due to the presence of competing hydroxylation of allylic C–H bonds. In addition, P450 monooxygenase-catalyzed epoxidation of olefins generally provides S -configured products with high enantiopurity, and examples of P450 enzymes demonstrating high R -enantioselectivity in epoxidation reactions remain rare. Herein, we report a chemoselective and enantiodivergent epoxidation of unactivated alkenes using engineered P450DA monooxygenases. The P450DA variants, obtained through structure-guided directed evolution based on the X-ray of P450DA-WT and P450DA-M3, switch the reactivity from the native hydroxylation of the allylic C–H bonds to epoxidation of C═C bonds and exhibit superior chemoselectivity (up to 99% epoxidation selectivity) and enantioselectivity (up to >99:1 er), delivering a wide variety of versatile and enantioenriched epoxides. Notably, an enantiodivergent synthesis was achieved simply by employing different P450DA variants, leading to both enantiomers of the epoxide products. Various transformations of the products were carried out, illustrating the synthetic utilities of the methods. Furthermore, molecular dockings and molecular dynamics simulations reveal the origin of high epoxidation selectivity and complementary stereoselectivity of the mutants.

A facile copper-catalyzed [3 + 2] cycloaddition of N -2,2,2-trifluoroethylisatin ketimines with various electron-deficient alkenes to access structurally polyfunctionalized spiro-pyrrolidine–oxindole motifs has been developed. Under the catalytic system, the N -2,2,2-trifluoroethylisatin ketimines could be utilized to react with a series of exocyclic alkenes, including 2-acylamino acrylates, 3-methylene-β-lactams, and sterically hindered cycloalkenes represented by cyclobutenone, to obtain a variety of densely functionalized spiro-pyrrolidine frameworks bearing an α-amino acid ester, β-lactam, and cyclobutanone, respectively, in generally good yields with excellent diastereo- and enantioselectivities.

N -Acyl/sulfonyl-α-functionalized 1,2,3,4-tetrahydroisoquinolines (THIQs) are significant structural motifs in organic synthesis and drug discovery. However, the one-pot approach enabling direct difunctionalization of THIQs remains challenging. Herein we report a photomediated one-pot three-component strategy to access N -acyl/sulfonyl-α-functionalized THIQs. This method features the use of oxygen (from air) as the green oxidant, high atom and step economy, and decent structural diversity. The synthetic applicability of the method was further demonstrated via the facile construction of valuable bioactive molecules. Mechanistic studies indicated that oxidation with singlet oxygen and the acceptor-less dehydrogenation were involved in the photoredox process.

Precisely controlling the product selectivity of a reaction is an important objective in organic synthesis. α-Ketoamides are vital intermediates in chemical transformations and privileged motifs in numerous drugs, natural products, and biologically active molecules. The selective synthesis of α-ketoamides from feedstock chemicals in a safe and operationally simple manner under mild conditions is a long-standing catalysis challenge. Herein, an unprecedented TBD-switched Pd-catalyzed double isocyanide insertion reaction for assembling ketoamides in aqueous DMSO from (hetero)aryl halides and pseudohalides under mild conditions is reported. The effectiveness and utility of this protocol are demonstrated by its diverse substrate scope (93 examples), the ability to late-stage modify pharmaceuticals, scalability to large-scale synthesis, and the synthesis of pharmaceutically active molecules. Mechanistic studies indicate that TBD is a key ligand that modulates the Pd-catalyzed double isocyanide insertion process, thereby selectively providing the desired α-ketoamides in a unique manner. In addition, the imidoylpalladium(II) complex and α-ketoimine amide are successfully isolated and determined by X-ray analysis, confirming that they are probable intermediates in the catalytic pathway.

Regioselective and enantioselective hydroxylation of propargylic C-H bonds are useful reactions but often lack appropriate catalysts. Here a green and efficient asymmetric hydroxylation of primary and secondary C-H bonds at propargylic positions has been established. A series of optically active propargylic alcohols were prepared with high regio- and enantioselectivity (up to 99% ee) under mild reaction conditions by using P450tol, while the C≡C bonds in the molecule remained unreacted. This protocol provides a green and practical method for constructing enantiomerically chiral propargylic alcohols. In addition, we also demonstrated that the biohydroxylation strategy was able to scaled up to 2.25 mmol scale with the production of chiral propargyl alcohol 2a at a yield of 196 mg with 96% ee, which's an important synthetic intermediate of antifungal drug Ravuconazole.