Paul Böhm


Last Name

Böhm

First Name

Paul

ORCID

0000-0003-0720-9766

Organisational unit

03980 - Piel, Jörn / Piel, Jörn

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Publications 1 - 2 of 2
  • Hijacking a bacterial ABC transporter for genetic code expansion
    Item type: Journal Article
    Iype , Tarun; Fottner, Maximilian; Böhm, Paul; et al. (2025)
    Nature
    The site-specific encoding of non-canonical amino acids (ncAAs) provides a powerful tool for expanding the functional repertoire of proteins1, 2, 3–4. Its widespread use for basic research and biotechnological applications is, however, hampered by the low efficiencies of current ncAA incorporation strategies. Here we reveal poor cellular ncAA uptake as a main obstacle to efficient genetic code expansion and overcome this bottleneck by hijacking a bacterial ATP-binding cassette (ABC) transporter5 to actively import easily synthesizable isopeptide-linked tripeptides that are processed into ncAAs within the cell. Using this approach, we enable efficient encoding of a variety of previously inaccessible ncAAs, decorating proteins with bioorthogonal6 and crosslinker7 moieties, post-translational modifications8,9 and functionalities for chemoenzymatic conjugation. We then devise a high-throughput directed evolution platform to engineer tailored transporter systems for the import of ncAAs that were historically refractory to efficient uptake. Customized Escherichia coli strains expressing these evolved transporters facilitate single and multi-site ncAA incorporation with wild-type efficiencies. Additionally, we adapt the tripeptide scaffolds for the co-transport of two different ncAAs, enabling their efficient dual incorporation. Collectively, our study demonstrates that engineering of uptake systems is a powerful strategy for programmable import of chemically diverse building blocks.
  • Genome-based discovery of polyketides generated by trans-acyltransferase polyketide synthases
    Item type: Book Chapter
    Lipsh-Sokolik, Rosalie; Böhm, Paul; Chepkirui, Clara; et al. (2025)
    Methods in Enzymology ~ Genomics-Guided Natural Products Discovery
    Polyketide synthases (PKSs) are multienzyme complexes that produce bioactive compounds with applications in medicine and agriculture. This review focuses on the genome-guided discovery of polyketides synthesized by trans-acyltransferase (trans-AT) PKSs, which, compared with the more prevalent cis-AT PKS counterparts, introduce distinct and complex chemical features that may entail specialized biological activities. With the rapid expansion of genomic data and abundance of trans-AT PKS biosynthetic gene clusters (BGCs), it is crucial to prioritize these clusters for experimental characterization based on their potential to produce novel molecules with valuable bioactivities. We outline methodologies for identifying BGCs, predicting the polyketide structure using ketosynthase phylogeny, and detail strategies for prioritizing BGCs for experimental characterization. We further describe approaches for producing extracts containing the target polyketide in the laboratory, as well as methods for its identification, isolation, and structural characterization. We expect that the strategies discussed in this review will accelerate the discovery of novel trans-AT PKS products, unlocking their potential to drive advancements in drug discovery.
Publications 1 - 2 of 2