Biology subjects

Wang, J.-b.

Publications and source records attributed to Wang, J.-b..

3 recordsLinked to original sources

bioRxiv2026

A High-Efficiency Geranylation Platform Enabled by Rational Tuning of a Single-Residue-Coded Chemo- and Regioselective Switch in Aromatic Prenyltransferases

Geranylation is an efficient post-modification strategy for enhancing or introducing new properties to molecules; however, achieving both high efficiency and precision remains challenging. Aromatic prenyltransferases (PTs) provide a mild and sustainable approach, yet their practical application is often limited by the trade-off between broad substrate scope and low selectivity, and successfully balancing these properties is rare. Here, using the pharmaceutically relevant cannabigerol framework as a model, we identify a single-residue switch in a prenyltransferase from Aspergillus terreus (AtaPT) that governs both chemo- and regioselectivity. Rational tuning of this switch preserves broad substrate scope while enhancing selectivity, enabling the selective formation of each of the three possible mono-geranylated products in the model reaction. Structural analysis reveals that this residue modulates the selectivity and activity through coupled steric and electrostatic effects. Notably, transplantation of this site into three homologous PTs similarly reprogrammed selectivity, establishing a conserved and generalizable control mechanism. Leveraging these insights, we achieved selective geranylation across 36 aromatic substrates, affording 41 products. Integration with an efficient geranyl donor regeneration system enabled preparative-scale synthesis, reaching titers up to 324 mg/L. This work defines a unifying mechanism for selectivity control in PTs and provides a high-efficiency platform for geranylation.

synthetic biology↗

bioRxiv2025

A De Novo Design Strategy to Convert FAcD from Dimer to Active Monomer

Enzymes largely exist in various oligomeric states, but monomeric enzymes are more conducive to industrial applications. Converting an oligomeric enzyme into an active monomer is a significant challenge. In this study, we present a de novo design strategy to convert fluoroacetate dehalogenase (FAcD) from its native dimeric form to an active monomer. Using the AI-based method ProteinMPNN, we identified critical protein-protein interaction (PPI) sites at the dimer interface. ArDCA, another AI tool, was employed to pinpoint catalytic hotspots. Six mutants, Mu1-Mu6, were designed. Molecular dynamics (MD) simulations, coupled with mass spectrometry, confirmed that these mutants form stable monomers. The pre-reaction state (PRS) model predicted that three of these mutants exhibited catalytic activity. In particular, Mu5 with 11 mutations from the wild-type, was predicted to have high catalytic activity, and was subsequently confirmed by kinetics experiment, with a kcat of 672.2 min-1 and a T5030 > 100 {degrees}C, comparable to the wild-type enzyme (kcat = 676.3 min-1, T5030 = 84 {degrees}C). Notably, the Y149M mutation increased catalytic activity nearly forty-fold, demonstrating the effectiveness of our design strategy.

bioengineering↗

bioRxiv2023

Metagenomic analysis of individual mosquitos reveals the ecology of insect viruses

Mosquito transmitted viruses are responsible for an increasing burden of human disease. Despite this, little is known about the diversity and ecology of viruses within individual mosquito hosts. Using a meta-transcriptomic approach, we analysed the virome of 2,438 individual mosquitos (79 species), spanning [~]4000 km along latitudes and longitudes in China. From these data we identified 393 core viral species associated with mosquitos, including seven (putative) arbovirus species. We identified potential species and geographic hotspots of viral richness and arbovirus occurrence, and demonstrated that host phylogeny had a strong impact on the composition of individual mosquito viromes. Our data revealed a large number of viruses shared among mosquito species or genera, expanding our knowledge of host specificity of insect-associated viruses. We also detected multiple virus species that were widespread throughout the country, possibly facilitated by long-distance mosquito migrations. Together, our results greatly expand the known mosquito virome, linked the viral diversity at the scale of individual insects to that at a country-wide scale, and offered unique insights into the ecology of viruses of insect vectors.

ecology↗