Prenylation of Flavanones by an Aromatic Prenyltransferase from <i>Fusarium globosum</i>

Prenylation increases the structural diversity and biological activity of flavonoids. In this study, an aromatic prenyltransferase, FgPT1, was identified from <i>Fusarium globosum</i>. This enzyme was demonstrated to specifically catalyze the prenylation of flavanones, including naringen...

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Main Authors: Dingtao Tang, Jiajie Quan, Zhengjiao Gao, Bingfeng He, Yu Hou, Peipei Fan, Meidong Pan, Jiali Yang
Format: Article
Language:English
Published: MDPI AG 2025-03-01
Series:Molecules
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Online Access:https://www.mdpi.com/1420-3049/30/7/1558
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author Dingtao Tang
Jiajie Quan
Zhengjiao Gao
Bingfeng He
Yu Hou
Peipei Fan
Meidong Pan
Jiali Yang
author_facet Dingtao Tang
Jiajie Quan
Zhengjiao Gao
Bingfeng He
Yu Hou
Peipei Fan
Meidong Pan
Jiali Yang
author_sort Dingtao Tang
collection DOAJ
description Prenylation increases the structural diversity and biological activity of flavonoids. In this study, an aromatic prenyltransferase, FgPT1, was identified from <i>Fusarium globosum</i>. This enzyme was demonstrated to specifically catalyze the prenylation of flavanones, including naringenin, hesperitin, eriodictyol, liquiritigenin, rac-pinocembrin, and dihydrogenistein, and exhibited no activity toward other types of flavonoids, including chalcones, flavonols, isoflavonoids, and flavonols. Ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) and nuclear magnetic resonance (NMR) analysis indicated that the majority of prenylated products were 6-<i>C</i> prenyl flavanones, with the exception of liquiritigenin, which was additionally transformed to 4′-<i>O</i> prenyl liquiritigenin. Enzyme kinetic analysis suggested that FgPT1 exhibited the highest catalytic efficiency towards naringenin, with a <i>k<sub>cat</sub></i>/<i>K<sub>M</sub></i> value determined as 61.92 s<sup>−1</sup> M<sup>−1</sup>, and the lowest catalytic efficiency towards liquiritigenin, with a <i>k<sub>cat</sub></i>/<i>K<sub>M</sub></i> of 1.18 s<sup>−1</sup> M<sup>−1</sup>. Biochemical characterization suggested that FgPT1 functioned as a metal-dependent enzyme with optimal activity in the presence of Ba<sup>2+</sup> at pH 7.5 and 30 °C. Site-directed mutagenesis resulted in a series of mutants, including A325V with impaired prenylation activity and V116I, V181I, and V194I with enhanced activity. V194I displayed the highest enzymatic activity with a nine-fold increase compared to wild-type FgPT1.
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spelling doaj-art-07cd2c85188f4ebd92a054b0e7be25e12025-08-20T02:09:17ZengMDPI AGMolecules1420-30492025-03-01307155810.3390/molecules30071558Prenylation of Flavanones by an Aromatic Prenyltransferase from <i>Fusarium globosum</i>Dingtao Tang0Jiajie Quan1Zhengjiao Gao2Bingfeng He3Yu Hou4Peipei Fan5Meidong Pan6Jiali Yang7School of Food Science and Engineering, Hainan University, Haikou 570228, ChinaSchool of Food Science and Engineering, Hainan University, Haikou 570228, ChinaSchool of Food Science and Engineering, Hainan University, Haikou 570228, ChinaSchool of Food Science and Engineering, Hainan University, Haikou 570228, ChinaSchool of Food Science and Engineering, Hainan University, Haikou 570228, ChinaSchool of Food Science and Engineering, Hainan University, Haikou 570228, ChinaSchool of Food Science and Engineering, Hainan University, Haikou 570228, ChinaSchool of Food Science and Engineering, Hainan University, Haikou 570228, ChinaPrenylation increases the structural diversity and biological activity of flavonoids. In this study, an aromatic prenyltransferase, FgPT1, was identified from <i>Fusarium globosum</i>. This enzyme was demonstrated to specifically catalyze the prenylation of flavanones, including naringenin, hesperitin, eriodictyol, liquiritigenin, rac-pinocembrin, and dihydrogenistein, and exhibited no activity toward other types of flavonoids, including chalcones, flavonols, isoflavonoids, and flavonols. Ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) and nuclear magnetic resonance (NMR) analysis indicated that the majority of prenylated products were 6-<i>C</i> prenyl flavanones, with the exception of liquiritigenin, which was additionally transformed to 4′-<i>O</i> prenyl liquiritigenin. Enzyme kinetic analysis suggested that FgPT1 exhibited the highest catalytic efficiency towards naringenin, with a <i>k<sub>cat</sub></i>/<i>K<sub>M</sub></i> value determined as 61.92 s<sup>−1</sup> M<sup>−1</sup>, and the lowest catalytic efficiency towards liquiritigenin, with a <i>k<sub>cat</sub></i>/<i>K<sub>M</sub></i> of 1.18 s<sup>−1</sup> M<sup>−1</sup>. Biochemical characterization suggested that FgPT1 functioned as a metal-dependent enzyme with optimal activity in the presence of Ba<sup>2+</sup> at pH 7.5 and 30 °C. Site-directed mutagenesis resulted in a series of mutants, including A325V with impaired prenylation activity and V116I, V181I, and V194I with enhanced activity. V194I displayed the highest enzymatic activity with a nine-fold increase compared to wild-type FgPT1.https://www.mdpi.com/1420-3049/30/7/1558enzyme catalysisflavanonesubstrate specificitysite-directed mutagenesis
spellingShingle Dingtao Tang
Jiajie Quan
Zhengjiao Gao
Bingfeng He
Yu Hou
Peipei Fan
Meidong Pan
Jiali Yang
Prenylation of Flavanones by an Aromatic Prenyltransferase from <i>Fusarium globosum</i>
Molecules
enzyme catalysis
flavanone
substrate specificity
site-directed mutagenesis
title Prenylation of Flavanones by an Aromatic Prenyltransferase from <i>Fusarium globosum</i>
title_full Prenylation of Flavanones by an Aromatic Prenyltransferase from <i>Fusarium globosum</i>
title_fullStr Prenylation of Flavanones by an Aromatic Prenyltransferase from <i>Fusarium globosum</i>
title_full_unstemmed Prenylation of Flavanones by an Aromatic Prenyltransferase from <i>Fusarium globosum</i>
title_short Prenylation of Flavanones by an Aromatic Prenyltransferase from <i>Fusarium globosum</i>
title_sort prenylation of flavanones by an aromatic prenyltransferase from i fusarium globosum i
topic enzyme catalysis
flavanone
substrate specificity
site-directed mutagenesis
url https://www.mdpi.com/1420-3049/30/7/1558
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