biostudies-literatureHan HFundamental Research Funds for the Central UniversitiesNatural Science Foundation of Heilongjiang Province of ChinaNational Natural Science Foundation of China275https://www.ebi.ac.uk/biostudies/studies/S-EPMC10965706biostudies-literatureUnknown108(1)Prenylation plays a pivotal role in the diversification and biological activities of natural products. This study presents the functional characterization of TolF, a multiple prenyltransferase from Tolypocladium inflatum. The heterologous expression of tolF in Aspergillus oryzae, coupled with feeding the transformed strain with paxilline, resulted in the production of 20- and 22-prenylpaxilline. Additionally, TolF demonstrated the ability to prenylated the reduced form of paxilline, β-paxitriol. A related prenyltransferase TerF from Chaunopycnis alba, exhibited similar substrate tolerance and regioselectivity. In vitro enzyme assays using purified recombinant enzymes TolF and TerF confirmed their capacity to catalyze prenylation of paxilline, β-paxitriol, and terpendole I. Based on previouApplied microbiology and biotechnologyBiochemical characterization of a multiple prenyltransferase from Tolypocladium inflatum.PMC10965706U22A2036931800031LH2023C0352572022BD0332370069Wang PWang QLi CQi JPeng SHan HWang HLiu CfalseBiochemical characterization of a multiple prenyltransferase from Tolypocladium inflatum.Prenylation plays a pivotal role in the diversification and biological activities of natural products. This study presents the functional characterization of TolF, a multiple prenyltransferase from Tolypocladium inflatum. The heterologous expression of tolF in Aspergillus oryzae, coupled with feeding the transformed strain with paxilline, resulted in the production of 20- and 22-prenylpaxilline. Additionally, TolF demonstrated the ability to prenylated the reduced form of paxilline, β-paxitriol. A related prenyltransferase TerF from Chaunopycnis alba, exhibited similar substrate tolerance and regioselectivity. In vitro enzyme assays using purified recombinant enzymes TolF and TerF confirmed their capacity to catalyze prenylation of paxilline, β-paxitriol, and terpendole I. Based on previou2024-01-01T00:00:00Z2024 Mar2026-06-03T06:52:15.905Z2025-04-04T22:59:33.649ZS-EPMC109657063853047010.1007/s00253-024-13113-6