Discovery of a gene cluster for the biosynthesis of novel cyclic peptide compound, KK-1, in Curvularia clavata
KK-1, a cyclic depsipeptide with 10 residues produced by a filamentous fungus Curvularia clavata BAUA-2787, is a promising pesticide active compound with high activity against many plant pathogens, especially Botrytis cinerea. As a first step toward the future mass production of KK-1 through synthet...
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2023-01-01
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| Series: | Frontiers in Fungal Biology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/ffunb.2022.1081179/full |
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| author | Shigenari Yamaguchi Tomonori Fujioka Akira Yoshimi Akira Yoshimi Toshitaka Kumagai Maiko Umemura Keietsu Abe Keietsu Abe Masayuki Machida Masayuki Machida Kiyoshi Kawai |
| author_facet | Shigenari Yamaguchi Tomonori Fujioka Akira Yoshimi Akira Yoshimi Toshitaka Kumagai Maiko Umemura Keietsu Abe Keietsu Abe Masayuki Machida Masayuki Machida Kiyoshi Kawai |
| author_sort | Shigenari Yamaguchi |
| collection | DOAJ |
| description | KK-1, a cyclic depsipeptide with 10 residues produced by a filamentous fungus Curvularia clavata BAUA-2787, is a promising pesticide active compound with high activity against many plant pathogens, especially Botrytis cinerea. As a first step toward the future mass production of KK-1 through synthetic biological approaches, we aimed to identify the genes responsible for the KK-1 biosynthesis. To achieve this, we conducted whole genome sequencing and transcriptome analysis of C. clavata BAUA-2787 to predict the KK-1 biosynthetic gene cluster. We then generated the overexpression and deletion mutants for each cluster gene using our originally developed transformation system for this fungus, and analyzed the KK-1 production and the cluster gene expression levels to confirm their involvement in KK-1 biosynthesis. As a result of these, a region of approximately 71 kb was found, containing 10 open reading frames, which were co-induced during KK-1 production, as a biosynthetic gene cluster. These include kk1B, which encodes nonribosomal peptide synthetase with a domain structure that is consistent with the structural features of KK-1, and kk1F, which encodes a transcription factor. The overexpression of kk1F increased the expression of the entire cluster genes and, consequently, improved KK-1 production, whereas its deletion decreased the expression of the entire cluster genes and almost eliminated KK-1 production, demonstrating that the protein encoded by kk1F regulates the expressions of the other nine cluster genes cooperatively as the pathway-specific transcription factor. Furthermore, the deletion of each cluster gene caused a reduction in KK-1 productivity, indicating that each gene is involved in KK-1 production. The genes kk1A, kk1D, kk1H, and kk1I, which showed a significant decrease in KK-1 productivity due to deletion, were presumed to be directly involved in KK-1 structure formation, including the biosynthesis of the constituent residues. kk1C, kk1E, kk1G, and kk1J, which maintained a certain level of KK-1 productivity despite deletion, were possibly involved in promoting or assisting KK-1 production, such as extracellular transportation and the removal of aberrant units incorporated into the peptide chain. |
| first_indexed | 2024-04-10T21:19:22Z |
| format | Article |
| id | doaj.art-2996407e834b4f28b3a2b9210ef402b6 |
| institution | Directory Open Access Journal |
| issn | 2673-6128 |
| language | English |
| last_indexed | 2024-04-10T21:19:22Z |
| publishDate | 2023-01-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Fungal Biology |
| spelling | doaj.art-2996407e834b4f28b3a2b9210ef402b62023-01-20T07:37:38ZengFrontiers Media S.A.Frontiers in Fungal Biology2673-61282023-01-01310.3389/ffunb.2022.10811791081179Discovery of a gene cluster for the biosynthesis of novel cyclic peptide compound, KK-1, in Curvularia clavataShigenari Yamaguchi0Tomonori Fujioka1Akira Yoshimi2Akira Yoshimi3Toshitaka Kumagai4Maiko Umemura5Keietsu Abe6Keietsu Abe7Masayuki Machida8Masayuki Machida9Kiyoshi Kawai10Biotechnology Laboratory, Life & Environment Research Center, Life Science Research Institute, Research & Development Division, Kumiai Chemical Industry Co., Ltd., Shizuoka, JapanBiotechnology Laboratory, Life & Environment Research Center, Life Science Research Institute, Research & Development Division, Kumiai Chemical Industry Co., Ltd., Shizuoka, JapanABE-Project, New Industry Creation Hatchery Center, Tohoku University, Sendai, JapanLaboratory of Terrestrial Microbial Ecology, Graduate School of Agriculture, Kyoto University, Kyoto, JapanFermlab Inc., Tokyo, JapanBio-system Research Group, Bioproduction Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba, JapanABE-Project, New Industry Creation Hatchery Center, Tohoku University, Sendai, JapanLaboratory of Applied Microbiology, Graduate School of Agricultural Science, Tohoku University, Sendai, JapanBio-system Research Group, Bioproduction Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba, JapanGraduate School of Engineering, Genome Biotechnology Laboratory, Kanazawa Institute of Technology, Ishikawa, JapanBiotechnology Laboratory, Life & Environment Research Center, Life Science Research Institute, Research & Development Division, Kumiai Chemical Industry Co., Ltd., Shizuoka, JapanKK-1, a cyclic depsipeptide with 10 residues produced by a filamentous fungus Curvularia clavata BAUA-2787, is a promising pesticide active compound with high activity against many plant pathogens, especially Botrytis cinerea. As a first step toward the future mass production of KK-1 through synthetic biological approaches, we aimed to identify the genes responsible for the KK-1 biosynthesis. To achieve this, we conducted whole genome sequencing and transcriptome analysis of C. clavata BAUA-2787 to predict the KK-1 biosynthetic gene cluster. We then generated the overexpression and deletion mutants for each cluster gene using our originally developed transformation system for this fungus, and analyzed the KK-1 production and the cluster gene expression levels to confirm their involvement in KK-1 biosynthesis. As a result of these, a region of approximately 71 kb was found, containing 10 open reading frames, which were co-induced during KK-1 production, as a biosynthetic gene cluster. These include kk1B, which encodes nonribosomal peptide synthetase with a domain structure that is consistent with the structural features of KK-1, and kk1F, which encodes a transcription factor. The overexpression of kk1F increased the expression of the entire cluster genes and, consequently, improved KK-1 production, whereas its deletion decreased the expression of the entire cluster genes and almost eliminated KK-1 production, demonstrating that the protein encoded by kk1F regulates the expressions of the other nine cluster genes cooperatively as the pathway-specific transcription factor. Furthermore, the deletion of each cluster gene caused a reduction in KK-1 productivity, indicating that each gene is involved in KK-1 production. The genes kk1A, kk1D, kk1H, and kk1I, which showed a significant decrease in KK-1 productivity due to deletion, were presumed to be directly involved in KK-1 structure formation, including the biosynthesis of the constituent residues. kk1C, kk1E, kk1G, and kk1J, which maintained a certain level of KK-1 productivity despite deletion, were possibly involved in promoting or assisting KK-1 production, such as extracellular transportation and the removal of aberrant units incorporated into the peptide chain.https://www.frontiersin.org/articles/10.3389/ffunb.2022.1081179/fullnon-ribosomal peptidebiosynthetic gene clusterantifungal agentgenome sequenceCurvularia clavatanatural fungicide |
| spellingShingle | Shigenari Yamaguchi Tomonori Fujioka Akira Yoshimi Akira Yoshimi Toshitaka Kumagai Maiko Umemura Keietsu Abe Keietsu Abe Masayuki Machida Masayuki Machida Kiyoshi Kawai Discovery of a gene cluster for the biosynthesis of novel cyclic peptide compound, KK-1, in Curvularia clavata Frontiers in Fungal Biology non-ribosomal peptide biosynthetic gene cluster antifungal agent genome sequence Curvularia clavata natural fungicide |
| title | Discovery of a gene cluster for the biosynthesis of novel cyclic peptide compound, KK-1, in Curvularia clavata |
| title_full | Discovery of a gene cluster for the biosynthesis of novel cyclic peptide compound, KK-1, in Curvularia clavata |
| title_fullStr | Discovery of a gene cluster for the biosynthesis of novel cyclic peptide compound, KK-1, in Curvularia clavata |
| title_full_unstemmed | Discovery of a gene cluster for the biosynthesis of novel cyclic peptide compound, KK-1, in Curvularia clavata |
| title_short | Discovery of a gene cluster for the biosynthesis of novel cyclic peptide compound, KK-1, in Curvularia clavata |
| title_sort | discovery of a gene cluster for the biosynthesis of novel cyclic peptide compound kk 1 in curvularia clavata |
| topic | non-ribosomal peptide biosynthetic gene cluster antifungal agent genome sequence Curvularia clavata natural fungicide |
| url | https://www.frontiersin.org/articles/10.3389/ffunb.2022.1081179/full |
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