Xiaoling Huang, Dan Li, Bi Long, Haidi Li, Jing Li, Wenxuan Wang, Kangping Xu and Xia Yu*,
{"title":"激活内生真菌 Talaromyces sp.的沉默基因簇,发现隐蔽的萘甲西隆代谢物。","authors":"Xiaoling Huang, Dan Li, Bi Long, Haidi Li, Jing Li, Wenxuan Wang, Kangping Xu and Xia Yu*, ","doi":"10.1021/acs.jafc.4c03162","DOIUrl":null,"url":null,"abstract":"<p >Fungal azaphilones have attracted widespread attention due to their significant potential as sources of food pigments and pharmaceuticals. Genome mining and gene cluster activation represent powerful tools and strategies for discovering novel natural products and bioactive molecules. Here, a putative azaphilone biosynthetic gene cluster <i>lut</i> from the endophytic fungus <i>Talaromyces</i> sp. was identified through genome mining. By overexpressing the pathway-specific transcription factor LutB, five new sclerotiorin-type azaphilones (<b>1</b>, <b>6</b>, <b>8</b>, and <b>10</b>–<b>11</b>) together with seven known analogues (<b>2</b>–<b>5</b>, <b>7</b>, <b>9</b>, <b>12</b>) were successfully produced. Compounds <b>8</b> and <b>9</b> exhibited antibacterial activity against <i>Bacillus subtilis</i> with MIC values of 64 and 16 μg/mL, respectively. Compound <b>11</b> showed cytotoxic activity against HCT116 and GES-1 with IC<sub>50</sub> values of 10.9 and 4.9 μM, respectively, while <b>1</b>, <b>4</b>, <b>5</b>, and <b>7</b>–<b>10</b> showed no obvious cytotoxic activity. Gene inactivation experiments confirmed the role of the <i>lut</i> cluster in the production of compounds <b>1</b>–<b>12</b>. Subsequent feeding experiments unveiled the novel functional diversity of the dual megasynthase system. Furthermore, a LutC-LutD binary oxidoreductase system was discovered, and in combination with DFT calculations, the basic biosynthetic pathway of the sclerotiorin-type azaphilones was characterized. This study provided a good example for the discovery of new azaphilones and further uncovered the biosynthesis of these compounds.</p>","PeriodicalId":41,"journal":{"name":"Journal of Agricultural and Food Chemistry","volume":null,"pages":null},"PeriodicalIF":5.7000,"publicationDate":"2024-07-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Activation of a Silent Gene Cluster from the Endophytic Fungus Talaromyces sp. Unearths Cryptic Azaphilone Metabolites\",\"authors\":\"Xiaoling Huang, Dan Li, Bi Long, Haidi Li, Jing Li, Wenxuan Wang, Kangping Xu and Xia Yu*, \",\"doi\":\"10.1021/acs.jafc.4c03162\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p >Fungal azaphilones have attracted widespread attention due to their significant potential as sources of food pigments and pharmaceuticals. Genome mining and gene cluster activation represent powerful tools and strategies for discovering novel natural products and bioactive molecules. Here, a putative azaphilone biosynthetic gene cluster <i>lut</i> from the endophytic fungus <i>Talaromyces</i> sp. was identified through genome mining. By overexpressing the pathway-specific transcription factor LutB, five new sclerotiorin-type azaphilones (<b>1</b>, <b>6</b>, <b>8</b>, and <b>10</b>–<b>11</b>) together with seven known analogues (<b>2</b>–<b>5</b>, <b>7</b>, <b>9</b>, <b>12</b>) were successfully produced. Compounds <b>8</b> and <b>9</b> exhibited antibacterial activity against <i>Bacillus subtilis</i> with MIC values of 64 and 16 μg/mL, respectively. Compound <b>11</b> showed cytotoxic activity against HCT116 and GES-1 with IC<sub>50</sub> values of 10.9 and 4.9 μM, respectively, while <b>1</b>, <b>4</b>, <b>5</b>, and <b>7</b>–<b>10</b> showed no obvious cytotoxic activity. Gene inactivation experiments confirmed the role of the <i>lut</i> cluster in the production of compounds <b>1</b>–<b>12</b>. Subsequent feeding experiments unveiled the novel functional diversity of the dual megasynthase system. Furthermore, a LutC-LutD binary oxidoreductase system was discovered, and in combination with DFT calculations, the basic biosynthetic pathway of the sclerotiorin-type azaphilones was characterized. This study provided a good example for the discovery of new azaphilones and further uncovered the biosynthesis of these compounds.</p>\",\"PeriodicalId\":41,\"journal\":{\"name\":\"Journal of Agricultural and Food Chemistry\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":5.7000,\"publicationDate\":\"2024-07-04\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Journal of Agricultural and Food Chemistry\",\"FirstCategoryId\":\"97\",\"ListUrlMain\":\"https://pubs.acs.org/doi/10.1021/acs.jafc.4c03162\",\"RegionNum\":1,\"RegionCategory\":\"农林科学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q1\",\"JCRName\":\"AGRICULTURE, MULTIDISCIPLINARY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of Agricultural and Food Chemistry","FirstCategoryId":"97","ListUrlMain":"https://pubs.acs.org/doi/10.1021/acs.jafc.4c03162","RegionNum":1,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"AGRICULTURE, MULTIDISCIPLINARY","Score":null,"Total":0}
Activation of a Silent Gene Cluster from the Endophytic Fungus Talaromyces sp. Unearths Cryptic Azaphilone Metabolites
Fungal azaphilones have attracted widespread attention due to their significant potential as sources of food pigments and pharmaceuticals. Genome mining and gene cluster activation represent powerful tools and strategies for discovering novel natural products and bioactive molecules. Here, a putative azaphilone biosynthetic gene cluster lut from the endophytic fungus Talaromyces sp. was identified through genome mining. By overexpressing the pathway-specific transcription factor LutB, five new sclerotiorin-type azaphilones (1, 6, 8, and 10–11) together with seven known analogues (2–5, 7, 9, 12) were successfully produced. Compounds 8 and 9 exhibited antibacterial activity against Bacillus subtilis with MIC values of 64 and 16 μg/mL, respectively. Compound 11 showed cytotoxic activity against HCT116 and GES-1 with IC50 values of 10.9 and 4.9 μM, respectively, while 1, 4, 5, and 7–10 showed no obvious cytotoxic activity. Gene inactivation experiments confirmed the role of the lut cluster in the production of compounds 1–12. Subsequent feeding experiments unveiled the novel functional diversity of the dual megasynthase system. Furthermore, a LutC-LutD binary oxidoreductase system was discovered, and in combination with DFT calculations, the basic biosynthetic pathway of the sclerotiorin-type azaphilones was characterized. This study provided a good example for the discovery of new azaphilones and further uncovered the biosynthesis of these compounds.
期刊介绍:
The Journal of Agricultural and Food Chemistry publishes high-quality, cutting edge original research representing complete studies and research advances dealing with the chemistry and biochemistry of agriculture and food. The Journal also encourages papers with chemistry and/or biochemistry as a major component combined with biological/sensory/nutritional/toxicological evaluation related to agriculture and/or food.