Jul 2026· Nature Chemical Biology· 7 citations· 51 references
Medicine
TL;DR
This work evolves enantioselective transcription factors that convert enzyme-catalyzed enantiomer concentrations into programmable gene expression outputs, focusing on imine reductases, and designs a genetic circuit that enables TF generation within weeks.
Abstract
Biocatalysts are prized for their enantioselectivity, but slow chromatographic separations required to measure enantiomeric excess bottleneck their development. To overcome this limitation, we evolve enantioselective transcription factors (eTFs) that convert enzyme-catalyzed enantiomer concentrations into programmable gene expression outputs, focusing on imine reductases. Here, using a massively parallel reporter assay, we measure dose-response curves for over 300,000 transcription factor variants in response to an imine precursor and chiral amine products. We quantify the sensitivity, selectivity and dynamic range across variants generated by random, site-saturation and shuffling mutagenesis, isolating variants with exceptional specificity. High-resolution structures of evolved eTFs elucidate how steric effects enforce enantioselectivity, while charge interactions distinguish the imine from the amines. Using two eTFs, we create an ultrahigh-throughput chiral screen to evolve an imine reductase with inverted enantioselectivity. To support generalizability and speed, we design a genetic circuit that enables TF generation within weeks. Our methods enable rapid measurement of asymmetric reactions, supporting innovation in chemical manufacturing.
This review systematically evaluates how machine learning integrates multimodal datasets, including sequence, structural, and functional performance data, to advance the discovery and engineering of naturally enantioselective enzymes, and enable the de novo design of artificial enzymes for reaction-relevant applications.
Jie Gu, Yan Xu, Xiao-Ge Sun et al.· ACS Synthetic Biology· 0 citations
Chiral nanoparticles offer new opportunities for designing functional materials with tunable optical and electrochemical properties. While cysteine and cysteine-containing peptides are commonly used as chiral agents in the synthesis of such nanoparticles, the use of alternative ligands remains limited. In this study, we introduce penicillamine as a new chiral agent in the seed-mediated growth of gold nanoparticles (AuNPs). The presence of L- or D-penicillamine induces chirality during nanoparticle growth, and its extent is examined at different growth stages using SEM. Structural evidence for enantioselective growth is also provided with high-resolution STEM. Circular dichroism spectroscopy of particle suspensions and dark-field scattering measurements at the single-particle level further suggest that the chirality is encoded in the nanoparticle morphology rather than being limited to surface-bound ligands. Importantly, dark-field scattering indicates that the AuNPs' chiral morphology is also preserved after drying on an Au electrode. To evaluate whether this morphological chirality translates into functional enantioselectivity, electrochemical measurements are performed using L- and D-cysteine as analytes on electrodes modified with chiral AuNPs. Cyclic voltammetry and electrochemical impedance spectroscopy show that electrodes modified with L-penicillamine-induced chiral AuNPs exhibit higher current densities (ca. 36%) and lower charge-transfer resistance toward L-cysteine oxidation. Similarly, D-penicillamine-induced chiral AuNPs show higher current densities for D-cysteine oxidation. This enantioselective interaction between enantio-matched pairs demonstrates that the encoded chirality influences molecular recognition processes at the electrode interface. Overall, this study establishes penicillamine-induced chiral AuNPs as versatile, label-free platforms for enantioselective electrochemical sensing.
The properties of polymers are governed by multi-level structural parameters, with tacticity being particularly critical for crystallization, thermal, and mechanical performance. Conventional approaches to modulate tacticity-such as tuning monomer enantiomeric excess or synthesizing distinct catalysts-are often complex and lack precise control. To address this challenge, we present a stereocontrol strategy by exploiting an enantiopure catalyst governed by an enantiomorphic site control (ESC) mechanism and polymeric exchange between the enantiomer pair. As a result, the enantiopure DiMeBiPh-BINOL-salen catalysts (R, R)-Y1 and (S, S)-Y1 were designed and demonstrated outstanding isoselectivity for the ROP of rac-BBL via the ESC mechanism. Their racemic pair rac-Y1 exhibited high syndioselectivity (Pr = 0.93) due to polymeric exchange. Ultimately, a series of P3HB products with streamlined tacticity (Pm: 0.07-0.90) was readily prepared by simply adjusting the enantiomeric excess (ee) values of the catalyst. A systematic investigation of the relationship between tacticity and material properties was performed. This work not only establishes an efficient and controllable route to tailor-made P3HB but also provides a scalable pathway for precise stereocontrol in polymer synthesis.
Min Xie, Jia-Hao Cui, W. Su et al.· Angewandte Chemie· 0 citations
Unnatural amino acids (UAAs) have become essential assets in the development of peptide- and peptidomimetic-based medications to enhance target selectivity and optimize physicochemical characteristics. The creation of peptides with drug-like qualities is still difficult, despite evidence that they may control receptor and alter disease processes. Herein, we report a visible light-driven enantioselective deaminative alkylation of glycine derivatives enabled by an in situ-generated chiral copper complex. The photoactive copper catalyst mediates single-electron reduction of Katritzky salts to generate alkyl radicals, which undergo stereocontrolled C(sp3)-C(sp3) bond formation. While deamination strategies are well explored, asymmetric variants remain underdeveloped. This methodology addresses that gap, delivering high enantioselectivity across a broad substrate scope, including natural product-derived and conformationally constrained scaffolds. The protocol is further applied to the synthesis of unnatural amino acid analogues and polypeptides without a loss of enantiopurity, demonstrating its utility and robustness in asymmetric synthesis.
Axially chiral biaryl dimethanols are ubiquitous and multifunctional intermediates for the synthesis of valuable atropisomeric molecules in advanced materials, drugs, natural products, and organocatalysts. Despite their broad synthetic utility, direct enantiodivergent catalytic access to this specific class of compounds remains largely unexplored, particularly in biocatalysis. Herein, we report a biocatalytic platform for enantiodivergent and atroposelective dynamic kinetic carbonyl reduction via transient seven-membered cyclic lactol intermediates. These engineered alcohol dehydrogenase (ADH)-driven transformations deliver up to 99% yield and enantioselectivity (>99:1 e.r. and < 1:99 e.r). This strategy demonstrates broad substrate compatibility, extending even to complementary “flipped” substrate series. Furthermore, the protocol is readily scalable to gram quantities and accommodates diverse downstream derivatizations. Mechanistic experiments and theoretical calculations delineate the pathway and illuminate the origin of enzymatic stereocontrol.
Jie Chen, Zhuoting Peng, Xiaolong Gao et al.· ACS Catalysis· 0 citations
The development of sustainable strategies for accessing optically pure amines is essential for pharmaceutical and fine‐chemical manufacturing. Here, we report the characterization of a newly identified imine reductase from
Goodfellowiella coeruleoviolacea
(GcIRED) and its application to the asymmetric reduction of cyclic imines in deep eutectic solvent based media. GcIRED displayed excellent catalytic performance in choline chloride/glycerol mixtures, enabling high substrate loadings up to 150 mM while maintaining high conversions and >99%
ee
toward (
S
)‐α‐substituted cyclic secondary amines. Compared to benchmark variants, GcIRED showed superior stability and activity in DES‐containing systems. A fed‐batch strategy further increased productivity, raising the turnover number from 8800 to 41 000 using a single enzyme and cofactor charge. Atomistic computational analyses revealed key NADPH‐anchoring residues and highlighted π−π and T‐shaped interactions between the substrate and the nicotinamide moiety, providing structural insight into productive binding and helping rationalize the catalytic behavior observed experimentally. Overall, this study establishes GcIRED as a robust and versatile biocatalyst for sustainable asymmetric amine synthesis and demonstrates how the integration of DES media can unlock improved performance in biocatalytic processes.
Federica De Nardi, Alessandro Marchetto, Nicoletta Schiavo et al.· Advanced Synthesis & Cat...· 0 citations
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