Site-selective dimethylation of flavonoids using fusion flavonoid O-methyltransferases
Abstract Flavonoids are often decorated with methyl groups, which are catalyzed by flavonoid O-methyltransferases (FOMTs). Most FOMTs methylate flavonoids in a regiospecific manner. Because of the regiospecific nature of FOMTs, the synthesis of polymethoxyflavonoids is accomplished by multiple O-met...
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2025-02-01
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| Series: | Applied Biological Chemistry |
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| Online Access: | https://doi.org/10.1186/s13765-025-00983-1 |
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| author | Kyungha Lee Seong Hee Bhoo Sang-Won Lee Man-Ho Cho |
| author_facet | Kyungha Lee Seong Hee Bhoo Sang-Won Lee Man-Ho Cho |
| author_sort | Kyungha Lee |
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| description | Abstract Flavonoids are often decorated with methyl groups, which are catalyzed by flavonoid O-methyltransferases (FOMTs). Most FOMTs methylate flavonoids in a regiospecific manner. Because of the regiospecific nature of FOMTs, the synthesis of polymethoxyflavonoids is accomplished by multiple O-methylation steps. The multistep synthesis of dimethoxyflavonoids can be efficiently performed by a one-pot procedure using a multienzyme biocatalyst. For the one-pot production of dimethoxyflavonoids, fusion FOMTs were generated by the combination of two different regiospecific FOMTs. RdOMT10 (flavonoid 3-OMT), OsNOMT (flavonoid 7-OMT), and ObFOMT5 (flavonoid 4'-OMT) were used in the FOMT fusion. The fusion FOMTs (OsNOMT/ObFOMT5, OsNOMT/RdOMT10, and ObFOMT5/RdOMT10) were heterologously expressed in Escherichia coli. Activity assays of the recombinant fusion FOMTs demonstrated that OsNOMT/ObFOMT5, OsNOMT/RdOMT10, and ObFOMT5/RdOMT10 catalyze 7/4'-O-methylations, 7/3-O-methylations, and 4'/3-O-methylations of flavonoids, respectively. OsNOMT/ObFOMT5 and OsNOMT/RdOMT10 showed strong dimethylation activity towards diverse flavonoids and were therefore used in the site-selective bioconversion of flavonoids into dimethoxyflavonoids. The E. coli cells bearing OsNOMT/ObFOMT5 successfully converted flavonoids into 7,4'-dimethoxyflavonoids. The engineered E. coli expressing OsNOMT/RdOMT10 converted flavonoids into 3,7-dimethoxyflavonoids. This result indicates that the fusion FOMTs are useful multienzyme biocatalysts for the site-selective production of dimethoxyflavonoids by one-pot bioconversion. |
| format | Article |
| id | doaj-art-cb09b0d870c14b4a838e228639a74268 |
| institution | Kabale University |
| issn | 2468-0842 |
| language | English |
| publishDate | 2025-02-01 |
| publisher | SpringerOpen |
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| series | Applied Biological Chemistry |
| spelling | doaj-art-cb09b0d870c14b4a838e228639a742682025-02-09T12:51:29ZengSpringerOpenApplied Biological Chemistry2468-08422025-02-0168111110.1186/s13765-025-00983-1Site-selective dimethylation of flavonoids using fusion flavonoid O-methyltransferasesKyungha Lee0Seong Hee Bhoo1Sang-Won Lee2Man-Ho Cho3Graduate School of Biotechnology, Kyung Hee UniversityDepartment of Genetics and Biotechnology, Kyung Hee UniversityDepartment of Genetics and Biotechnology, Kyung Hee UniversityDepartment of Genetics and Biotechnology, Kyung Hee UniversityAbstract Flavonoids are often decorated with methyl groups, which are catalyzed by flavonoid O-methyltransferases (FOMTs). Most FOMTs methylate flavonoids in a regiospecific manner. Because of the regiospecific nature of FOMTs, the synthesis of polymethoxyflavonoids is accomplished by multiple O-methylation steps. The multistep synthesis of dimethoxyflavonoids can be efficiently performed by a one-pot procedure using a multienzyme biocatalyst. For the one-pot production of dimethoxyflavonoids, fusion FOMTs were generated by the combination of two different regiospecific FOMTs. RdOMT10 (flavonoid 3-OMT), OsNOMT (flavonoid 7-OMT), and ObFOMT5 (flavonoid 4'-OMT) were used in the FOMT fusion. The fusion FOMTs (OsNOMT/ObFOMT5, OsNOMT/RdOMT10, and ObFOMT5/RdOMT10) were heterologously expressed in Escherichia coli. Activity assays of the recombinant fusion FOMTs demonstrated that OsNOMT/ObFOMT5, OsNOMT/RdOMT10, and ObFOMT5/RdOMT10 catalyze 7/4'-O-methylations, 7/3-O-methylations, and 4'/3-O-methylations of flavonoids, respectively. OsNOMT/ObFOMT5 and OsNOMT/RdOMT10 showed strong dimethylation activity towards diverse flavonoids and were therefore used in the site-selective bioconversion of flavonoids into dimethoxyflavonoids. The E. coli cells bearing OsNOMT/ObFOMT5 successfully converted flavonoids into 7,4'-dimethoxyflavonoids. The engineered E. coli expressing OsNOMT/RdOMT10 converted flavonoids into 3,7-dimethoxyflavonoids. This result indicates that the fusion FOMTs are useful multienzyme biocatalysts for the site-selective production of dimethoxyflavonoids by one-pot bioconversion.https://doi.org/10.1186/s13765-025-00983-1DimethoxyflavonoidFlavonoid O-methyltransferaseFusion FOMTOne-pot bioconversion |
| spellingShingle | Kyungha Lee Seong Hee Bhoo Sang-Won Lee Man-Ho Cho Site-selective dimethylation of flavonoids using fusion flavonoid O-methyltransferases Applied Biological Chemistry Dimethoxyflavonoid Flavonoid O-methyltransferase Fusion FOMT One-pot bioconversion |
| title | Site-selective dimethylation of flavonoids using fusion flavonoid O-methyltransferases |
| title_full | Site-selective dimethylation of flavonoids using fusion flavonoid O-methyltransferases |
| title_fullStr | Site-selective dimethylation of flavonoids using fusion flavonoid O-methyltransferases |
| title_full_unstemmed | Site-selective dimethylation of flavonoids using fusion flavonoid O-methyltransferases |
| title_short | Site-selective dimethylation of flavonoids using fusion flavonoid O-methyltransferases |
| title_sort | site selective dimethylation of flavonoids using fusion flavonoid o methyltransferases |
| topic | Dimethoxyflavonoid Flavonoid O-methyltransferase Fusion FOMT One-pot bioconversion |
| url | https://doi.org/10.1186/s13765-025-00983-1 |
| work_keys_str_mv | AT kyunghalee siteselectivedimethylationofflavonoidsusingfusionflavonoidomethyltransferases AT seongheebhoo siteselectivedimethylationofflavonoidsusingfusionflavonoidomethyltransferases AT sangwonlee siteselectivedimethylationofflavonoidsusingfusionflavonoidomethyltransferases AT manhocho siteselectivedimethylationofflavonoidsusingfusionflavonoidomethyltransferases |