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High-Throughput Profiling of Prenylelongases Enables the Assembly of Modified Prenoids
Hui Li1, Anjali Sital1, Clemens Mayer1
1Biomolecular Chemistry and Catalysis, Stratingh Institute for Chemistry, University of Groningen, Nijenborgh 4, 9747 AG Groningen, The Netherlands.
ACS Chemical Biology
|June 15, 2026
Summary
This study introduces a novel high-throughput assay for prenylelongases (PEs) using accessible monophosphates, overcoming challenges with pyrophosphate substrates. This facilitates biocatalytic production of modified prenoids for diverse applications.
Area of Science:
- Biocatalysis and Enzyme Engineering
- Synthetic Biology
- Chemical Biology
Background:
- Modified prenoids are valuable in chemical biology, fragrances, and bioactive compound synthesis.
- Current synthetic routes are laborious, limiting widespread application of modified prenoids.
- Prenylelongases (PEs) offer biocatalytic potential but are hindered by difficult assays and expensive pyrophosphate substrates.
Purpose of the Study:
- To develop a high-throughput assay for prenylelongases (PEs) using readily available monophosphate substrates.
- To enable efficient biochemical characterization and engineering of PEs for expanded substrate scopes.
- To facilitate biocatalytic production of modified prenoids and their derivatives.
Main Methods:
- Developed continuous high-throughput assays for PEs utilizing monophosphates instead of pyrophosphates.
- Biochemically characterized a panel of diverse PEs and profiled their activity with non-native substrates.
- Conducted two exploratory enzyme engineering campaigns and demonstrated preparative-scale synthesis of modified prenoids.
Main Results:
- Successfully established a high-throughput assay platform for PEs, bypassing the need for costly pyrophosphate synthesis.
- Characterized multiple PEs, revealing their substrate specificities and potential for modification.
- Demonstrated the utility of engineered PEs for selective, (semi)preparative scale synthesis of modified prenoids.
Conclusions:
- The developed monophosphate-based assay simplifies PE interrogation and engineering.
- This platform facilitates biocatalytic access to a wider range of modified prenoids.
- The findings pave the way for broader applications of engineered PEs in chemical synthesis.
