1. Retrobiosynthesis
Previous publications: Nature Catalysis 2025
We recently completed digitally recording tens of thousands of reactions from modular polyketide synthases (PKSs)/nonribosomal peptide synthetases (NRPSs), and established the ClusterCAD database along with the open-source automated design tool ClusterCAD RetroTide. This tool can automatically design and optimize the selection of PKS/NRPS modules based on the target product structure. To practice ClusterCAD RetroTide, I first used simple but industrially important targets (diols and amino alcohols) that require only two PKS modules for synthesis. Using technologies in metabolic engineering, enzyme mining, structural biology and automated computational design, we integrated versatile PKS initiation and reductive termination strategies into designed pathways. As a result, 17 industrial diols and derivatives were biosynthesized in harnessed Streptomyces chassis at high titers, demonstrating the feasibility of designing such a biomanufacturing platform.
Our future goal is to systematically overcome the challenges of PKS/NRPS engineering to complete “a thousand molecule project”. With sophisticated retrosynthesis tools and harnessed hosts, we plan to achieve the rapid and efficient assembly of 5–10 PKS/NRPS modules and biosynthesize over 1,000 new-to-nature small molecules in the short term and 10,000 molecules in the long term.
2. Enzymology in Natural Product Biosynthesis
Previous publications: Nature Chemistry 2019
Fungal indole alkaloids are of significant interest due to their broad biological activities, including calmodulin-inhibitory malbrancheamides and anthelmintic paraherquamides. We elucidated the malbrancheamide biosynthetic pathway using biomimetic total syntheses to generate racemic alkaloids and intermediates, along with in vitro enzymatic reconstitution to yield the natural antipode (+)-malbrancheamide. Our work revealed a common NRPS biogenetic scheme involving dipeptide reductive cleavage and a cascade of reactions leading to intramolecular [4 + 2] hetero Diels-Alder (IMDA) cyclization. The bifunctional enzyme MalC, a reductase/Diels-Alderase, is crucial for synthesizing optically pure (+)-malbrancheamide with strict stereocontrol, and we determined the structural basis for IMDA catalysis. Taken together, these new discoveries provide a novel enzymatic toolkit for developing improved anthelmintic therapeutics to address human and animal diseases.
Reducing carbon emissions from aviation transportation sectors requires the development of sustainable jet fuels. We engineered iterative polyketide synthase (iPKS) pathways to produce polycyclopropanated fatty acids (POP-FAs), a new class of energy-dense molecules with potential applications as sustainable aviation and rocket fuels. Furthermore, by combining computational fuel-property modeling, genome mining and pathway engineering, we identified cyclopropanases as a key iPKS component controlling both carbon-chain length and cyclopropane ratio, allowing precise tuning of fuel structure, energy density, and predicted freezing-point properties. This work establishes a versatile biosynthetic platform for producing customizable high-performance fuels from renewable carbon sources and provides new design principles for engineering iterative polyketide synthases.
In the future, we plan to enable many major types of enzyme decoration onto common PKS/NRPS core structures, featuring programmable access to precisely tuned chemistry. Using the ESM language model, which captures sequence, structural, and functional aspects of protein evolution, we will also develop low-N evolution strategies that only require a small number (“N”) of experimentally tested variants to achieve improved or even new protein functions.
3. Metabolic Engineering
Previous publications: Coming soon
Over the past few decades, bioorthogonal chemistry, which allows chemistry to be installed in microbes without disrupting host metabolism, has transformed the interface between chemistry and biology. In parallel, advances in directed evolution and genome mining have vastly expanded chemical reactions available within living systems. The full integration of these once “abiotic” reactions into cellular metabolism has now become a rapidly emerging field, and it raises a fundamental question: how do microbes respond to the new reactions we engineer them to perform? The accumulated knowledge will contribute to the development of improved microbial hosts.
We began to explore this question using our diol biosynthetic pipelines, where designed unnatural PKSs produce large quantities of aldehyde intermediates that are highly toxic to host cells. This stress resulted in the accumulation of hundreds of genome-wide mutations. Through detailed investigations, we orchestrated PKS pathway design to mitigate the cellular burden; as a result, we largely lowered mutation rates and achieved more than ten-fold increase in diol titers.