Biosynthesis of lactacystin as a proteasome inhibitor
Abstract Lactacystin is an irreversible proteasome inhibitor isolated from Streptomyces lactacystinicus. Despite its importance for its biological activity, the biosynthesis of lactacystin remains unknown. In this study, we identified the lactacystin biosynthetic gene cluster by gene disruption and...
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Nature Portfolio
2025-01-01
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| Series: | Communications Chemistry |
| Online Access: | https://doi.org/10.1038/s42004-025-01406-4 |
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| author | Takeshi Tsunoda Shunkichi Furumura Haruka Yamazaki Chitose Maruyama Yoshimitsu Hamano Yasushi Ogasawara Tohru Dairi |
| author_facet | Takeshi Tsunoda Shunkichi Furumura Haruka Yamazaki Chitose Maruyama Yoshimitsu Hamano Yasushi Ogasawara Tohru Dairi |
| author_sort | Takeshi Tsunoda |
| collection | DOAJ |
| description | Abstract Lactacystin is an irreversible proteasome inhibitor isolated from Streptomyces lactacystinicus. Despite its importance for its biological activity, the biosynthesis of lactacystin remains unknown. In this study, we identified the lactacystin biosynthetic gene cluster by gene disruption and heterologous expression experiments. We also examined the functions of the genes encoding a PKS/NRPS hybrid protein (LctA), NRPS (LctB), ketosynthase-like cyclase (LctC), cytochrome P450 (LctD), MbtH-like protein (LctE), and formyltransferase (LctF) by in vivo and in vitro experiments. In particular, we demonstrated that LctF directly transferred the formyl group of 10-N-formyl tetrahydrofolate to CoA. The formyl group of formyl-CoA was then transferred to ACP1 by LctA_AT1 to form formyl-ACP1. This is the first example of an AT domain recognizing a formyl group. The formyl group is perhaps transferred to methylmalonate tethered on LctA_ACP2 to yield methylmalonyl-semialdehyde-ACP2. Then, it would be condensed with leucine bound to PCP in LctB by the C domain in LctA. Using a mimic compound, we confirmed that LctC catalyzed the formation of the cyclic α,α-disubstituted amino acid structure with concomitant release of the product from PCP. Thus, we figured out the overall biosynthesis of lactacystin including a novel role of a formyl group in a secondary metabolite. |
| format | Article |
| id | doaj-art-7152d142ee7440b39983074ec7cd44bb |
| institution | Kabale University |
| issn | 2399-3669 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Communications Chemistry |
| spelling | doaj-art-7152d142ee7440b39983074ec7cd44bb2025-01-19T12:13:12ZengNature PortfolioCommunications Chemistry2399-36692025-01-01811810.1038/s42004-025-01406-4Biosynthesis of lactacystin as a proteasome inhibitorTakeshi Tsunoda0Shunkichi Furumura1Haruka Yamazaki2Chitose Maruyama3Yoshimitsu Hamano4Yasushi Ogasawara5Tohru Dairi6Graduate School of Engineering, Hokkaido University, N13-W8, Kita-ku, SapporoGraduate School of Chemical Sciences and Engineering, Hokkaido University, N13-W8, Kita-ku, SapporoGraduate School of Chemical Sciences and Engineering, Hokkaido University, N13-W8, Kita-ku, SapporoGraduate School of Bioscience and Biotechnology, Fukui Prefectural University, Eiheiji-choGraduate School of Bioscience and Biotechnology, Fukui Prefectural University, Eiheiji-choGraduate School of Engineering, Hokkaido University, N13-W8, Kita-ku, SapporoGraduate School of Engineering, Hokkaido University, N13-W8, Kita-ku, SapporoAbstract Lactacystin is an irreversible proteasome inhibitor isolated from Streptomyces lactacystinicus. Despite its importance for its biological activity, the biosynthesis of lactacystin remains unknown. In this study, we identified the lactacystin biosynthetic gene cluster by gene disruption and heterologous expression experiments. We also examined the functions of the genes encoding a PKS/NRPS hybrid protein (LctA), NRPS (LctB), ketosynthase-like cyclase (LctC), cytochrome P450 (LctD), MbtH-like protein (LctE), and formyltransferase (LctF) by in vivo and in vitro experiments. In particular, we demonstrated that LctF directly transferred the formyl group of 10-N-formyl tetrahydrofolate to CoA. The formyl group of formyl-CoA was then transferred to ACP1 by LctA_AT1 to form formyl-ACP1. This is the first example of an AT domain recognizing a formyl group. The formyl group is perhaps transferred to methylmalonate tethered on LctA_ACP2 to yield methylmalonyl-semialdehyde-ACP2. Then, it would be condensed with leucine bound to PCP in LctB by the C domain in LctA. Using a mimic compound, we confirmed that LctC catalyzed the formation of the cyclic α,α-disubstituted amino acid structure with concomitant release of the product from PCP. Thus, we figured out the overall biosynthesis of lactacystin including a novel role of a formyl group in a secondary metabolite.https://doi.org/10.1038/s42004-025-01406-4 |
| spellingShingle | Takeshi Tsunoda Shunkichi Furumura Haruka Yamazaki Chitose Maruyama Yoshimitsu Hamano Yasushi Ogasawara Tohru Dairi Biosynthesis of lactacystin as a proteasome inhibitor Communications Chemistry |
| title | Biosynthesis of lactacystin as a proteasome inhibitor |
| title_full | Biosynthesis of lactacystin as a proteasome inhibitor |
| title_fullStr | Biosynthesis of lactacystin as a proteasome inhibitor |
| title_full_unstemmed | Biosynthesis of lactacystin as a proteasome inhibitor |
| title_short | Biosynthesis of lactacystin as a proteasome inhibitor |
| title_sort | biosynthesis of lactacystin as a proteasome inhibitor |
| url | https://doi.org/10.1038/s42004-025-01406-4 |
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