The utility of Streptococcus mutans undecaprenol kinase for the chemoenzymatic synthesis of diverse non-natural isoprenoids

• Published in: Bioorganic chemistry 2024-10, Vol.151, p.107707, Article 107707
• Main Authors: Kumar, Vikas, Johnson, Bryce P., Mandal, Prashant S., Sheffield, Daniel R., Dimas, Dustin A., Das, Riki, Maity, Sanjay, Distefano, Mark D., Singh, Shanteri
• Format: Article
• Language: English
• Subjects: Biotechnology; Enzyme cascade; Isopentenyl phosphate kinase; Isoprenoids; Molecular Structure; Streptococcus mutans - enzymology; Terpenes - chemical synthesis; Terpenes - chemistry; Terpenes - metabolism; Undecaprenol kinase
• ISSN: 0045-2068; 1090-2120; 1090-2120
• DOI: 10.1016/j.bioorg.2024.107707

[Display omitted] •The promiscuity of Streptococcus mutans undecaprenol kinase extends to alcohols as small as 4 carbons.•Undecaprenol kinases can be coupled with downstream enzymes from isoprenoid pathways.•An undecaprenol-kinase-based enzyme cascade can generate novel isoprenoids, opening up avenues for synthetic biology. Isoprene chemoenzymatic cascades (ICCs) overcome the complexity of natural pathways by leveraging a streamlined two-enzyme cascade, facilitating efficient synthesis of C5-isoprene diphosphate precursors from readily available alcohol derivatives. Despite the documented promiscuity of enzymes in ICCs, exploration of their potential for accessing novel compounds remains limited, and existing methods require additional enzymes for generating longer-chain diphosphates. In this study, we present the utility of Streptococcus mutans undecaprenol kinase (SmUdpK) for the chemoenzymatic synthesis of diverse non-natural isoprenoids. Using a library of 50 synthetic alcohols, we demonstrate that SmUdpK’s promiscuity extends to allylic chains as small as four carbons and benzylic alcohols with various substituents. Subsequently, SmUdpK is utilized in an ICC with isopentenyl phosphate kinase and aromatic prenyltransferase to generate multiple non-natural isoprenoids. This work provides evidence that, with proper optimization, SmUdpK can act as the first enzyme in these ICCs, enhancing access to both valuable and novel compounds.