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Aug 2026

Post-translational chemical modification of E3 ligase for efficient target protein degradation.

This work reports the first ligand-directed chemical strategy that converts transient PROTAC-mediated ternary complex formation into binary target recognition via post-translational chemical modification of an E3 ligase, and believes it could provide a platform for next-generation targeted protein degraders to overcome the current limitation of PROTAC approach.

Eunbin Park, Jinjoo Jung, Gangasani Jagadeesh Kumar et al. · 0 citations
Open access Jul 2026

A tailored CoA ligase–N-acyltransferase cascade for on-DNA amide bond formation gives access to broad substrate scope

DNA-encoded chemical library (DEL) technology is a powerful tool in early-stage drug discovery. Although widely applied in industry and academia, challenges persist in generating DELs with high quality and chemical diversity. Low yields in building-block incorporation, limited selectivity and, most importantly, DNA damage from harsh reaction conditions compromise library quality, reduce signal-to-noise in affinity selections and ultimately hinder drug discovery. Here we show that tailored enzymes can be harnessed for the effective construction of molecular diversity on DNA under mild conditions. Targeting amide bond formation, we designed a cascade of complementary coenzyme A ligases and rationally tailored N-acyltransferases to access a broad amide scope on-DNA (>120 examples), identifying structural elements that optimize the biocatalysts’ DNA compatibility in the process. Integrating the enzymatic cascade with chemical synthesis led to the construction of a diverse DEL without damage to the DNA barcode, highlighting the biocatalysts’ applicability for early scaffold construction and late-stage functionalization. DNA-encoded library (DEL) synthesis can be constrained by DNA damage and limited reaction scope under conventional chemistry. Here a complementary CoA ligase–N-acyltransferase cascade is engineered to enable high yield, DNA-compatible amide formation, expanding substrate scope and DEL quality.

Daniel Schaub, Alice Lessing, Fabian Meyer et al. · 1 citation
Jul 2026

Overcoming catalytic barriers to reprogram acyltransferases for bis(2-hydroxyethyl) terephthalate hydrolysis.

The extensive use of polyethylene terephthalate (PET) has resulted in severe environmental pollution and ecological stress. Despite advances in PET recycling, current processes struggle to achieve high product value, as the complete conversion to terephthalic acid remains energetically demanding and economically inefficient. The catalytic promiscuity inherent in natural enzyme evolution holds great promise for providing novel candidates to accelerate PET biodegradation and upcycling. Herein, we report for the first time that the acyltransferase metA from Mycobacterium tuberculosis (MtMetA) catalyzes the conversion of bis(2-hydroxyethyl) terephthalate (BHET), an intermediate of PET hydrolysis, into the high-value monomer mono(2-hydroxyethyl) terephthalate (MHET). Guided by molecular dynamics (MD) simulations, we applied a catalytic barrier-minimization strategy to optimize the active-site environment of MtMetA, yielding engineered variants, notably ΔBarrier2 and ΔBarrier3. Specifically, ΔBarrier2 achieved a 3.1-fold increase in MHET yield, while ΔBarrier3 demonstrated a 3.2-fold enhancement in catalytic efficiency (kcat/KM) relative to the wild-type. The truncated MtMetA variants also exhibited enhanced robustness, showing improved thermostability (3.5-fold increase in residual activity at 60 °C for ΔBarrier3), as well as higher tolerance to metal ions and organic solvents. Specifically, ΔBarrier2 displayed a 7.2-fold increase in product yield in Ca2+-containing systems, while ΔBarrier3 retained 2.3-fold higher residual activity in the presence of 50 % (v/v) isopropanol. MD simulations revealed that an enlarged active pocket and a shortened nucleophilic attack distance synergistically govern the enhanced catalytic activity and robustness. This work expands the enzymatic toolbox for PET recycling and targeted BHET degradation, advancing sustainable plastic waste management through biocatalytic innovations.

Jie Qiao, Yibo Song, Nan Zhao et al. · 0 citations
Aug 2026

Improved Purification of sgRNA and pegRNA by Use of a 5'-Terminal Dioctyloxytrityl (DOT) Protecting Group.

The purity and chemical integrity of oligonucleotides are critical determinants of safety, efficacy, and reproducibility in RNA therapeutics. Impurities arising from incomplete synthesis, degradation, or side reactions can compromise pharmacological performance and induce off-target or immune responses. For long, structured oligonucleotides such as CRISPR guides, purification can be costly in both resources and yield. To overcome these limitations, we developed a purification method based on a 5'-terminal dioctyloxytrityl (DOT) protecting group. In this purification method, DOT-protected full-length compounds show high-resolution separation from truncated impurities. The DOT group can then be removed under mild, rapid, and scalable conditions. DOT chemistry is analogous to "DMT-ON" approaches that are well-known and widely used in the field, and is fully compatible with existing synthesis workflows, but dramatically improves HPLC purification relative to DMT-ON approaches by providing higher retention and thus better resolution to enable the purification of long, complex, and chemically modified sequences.

Atish Wagh, Jonathan K. Watts · 0 citations
Open access Aug 2026

Directed Evolution of a Highly Selective Fungal Peroxygenase for the Synthesis of Indigo

The large-scale production of indigo, a widely used textile dye, currently relies on chemical processes that consume non-renewable resources and that generate toxic waste, although biocatalysis is emerging as an environmentally friendly alternative. Here, an unspecific peroxygenase from the fungus Daldinia sp. EC12 (DspUPO-I) was subjected to a directed evolution campaign for activity and expression in yeast. With four mutations in the mature protein, the resulting evolved variant showed 95% regioselectivity for indoxyl formation, the precursor of indigo, along with a striking resistance to oxidative inactivation by hydrogen peroxide. These features enable this biocatalyst to synthesize 0.70 g/L indigo on a semipreparative scale. Molecular dynamics simulations highlighted the particular geometry of the broad heme-access channel as the main driving force for the high selectivity and the oxidative stability of this robust biocatalyst during indigo production.

Alejandro Beltrán-Nogal, Ivan Mateljak, Dianelis T. Monterrey et al. · 0 citations
Open access Aug 2026

Sequence-Guided Engineering of a Bacterial Diterpene Synthase Induces Further Cyclization

Enzyme engineering serves as a powerful tool in biocatalysis, enabling the development of enzymes with improved stability, activity, and specificity for a range of academic, industrial, and pharmaceutical applications. However, a limited understanding of sequence–structure–function relationships in terpene synthases, the enzymes that form the complex polycyclic hydrocarbon skeletons of terpenoid natural products, presents a major challenge in predicting and engineering the products of terpene synthases. In this study, we investigated the product profiles of two bacterial variediene synthases, OdVS from Olivibacter domesticus and PsVS from Prauserella shujinwangii, and found that they share some products with the bacterial phomopsene synthase, PmS from Allokutzneria albata, despite low sequence similarity. Thirteen diterpenes were isolated and structurally elucidated including two previously unreported compounds. A series of variants of OdVS, PsVS, and PmS were constructed by targeting conserved residues around the active site and aided in the identification of key residues that control the cyclization pathway. Ultimately, mutation of a single residue, PsVSY86L, was found to switch the major product of PsVS from the tricyclic variediene to the tetracyclic phomopsene, although this switch came at the cost of significantly reduced overall yield indicating a tradeoff between activity and product diversification.

Xiu-Ting Wei, Wenbo Ning, Pin-Shuo Huang et al. · 0 citations