Strategy of employing plug-and-play vectors and LC–MS screening to facilitate the discovery of natural products using Aspergillus oryzae
Abstract Aspergillus oryzae is a widely used host for heterologous expression of fungal natural products. However, the vectors previously developed are not convenient for use and screening positive transformants by PCR and fermentation is time- and effort-consuming. Hence, three plug-and-play vector...
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| Format: | Article |
| Language: | English |
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SpringerOpen
2025-01-01
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| Series: | Bioresources and Bioprocessing |
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| Online Access: | https://doi.org/10.1186/s40643-024-00833-w |
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| author | Hanliang Shi Beibei Lin Mengmeng Zheng Fengyu Gan Zhi Lin Xiujuan Xin Jian Zhao Xudong Qu Faliang An |
| author_facet | Hanliang Shi Beibei Lin Mengmeng Zheng Fengyu Gan Zhi Lin Xiujuan Xin Jian Zhao Xudong Qu Faliang An |
| author_sort | Hanliang Shi |
| collection | DOAJ |
| description | Abstract Aspergillus oryzae is a widely used host for heterologous expression of fungal natural products. However, the vectors previously developed are not convenient for use and screening positive transformants by PCR and fermentation is time- and effort-consuming. Hence, three plug-and-play vectors were developed here for multi-gene expression and liquid chromatography mass spectrometry detection was introduced to screen positive transformants. Using rug BGC for verification, we demonstrated that the vectors we developed perform well and liquid chromatography mass spectrometry detection is feasible to screen positive transformants. For deleterious gene expression, PxyrA rather than PamyB was employed. Utilizing the toolkit described here to express natural products, dozen days can be saved. Graphical Abstract |
| format | Article |
| id | doaj-art-ceec33fc494248cfb9aaeb3e0e208de0 |
| institution | Kabale University |
| issn | 2197-4365 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | SpringerOpen |
| record_format | Article |
| series | Bioresources and Bioprocessing |
| spelling | doaj-art-ceec33fc494248cfb9aaeb3e0e208de02025-01-12T12:06:02ZengSpringerOpenBioresources and Bioprocessing2197-43652025-01-0112111110.1186/s40643-024-00833-wStrategy of employing plug-and-play vectors and LC–MS screening to facilitate the discovery of natural products using Aspergillus oryzaeHanliang Shi0Beibei Lin1Mengmeng Zheng2Fengyu Gan3Zhi Lin4Xiujuan Xin5Jian Zhao6Xudong Qu7Faliang An8State Key Laboratory of Bioreactor Engineering, East China University of Science and TechnologyState Key Laboratory of Bioreactor Engineering, East China University of Science and TechnologyState Key Laboratory of Microbial Metabolism, School of Life Sciences and Biotechnology, Shanghai Jiao Tong UniversityState Key Laboratory of Bioreactor Engineering, East China University of Science and TechnologyState Key Laboratory of Microbial Metabolism, School of Life Sciences and Biotechnology, Shanghai Jiao Tong UniversityState Key Laboratory of Bioreactor Engineering, East China University of Science and TechnologyState Key Laboratory of Bioreactor Engineering, East China University of Science and TechnologyState Key Laboratory of Microbial Metabolism, School of Life Sciences and Biotechnology, Shanghai Jiao Tong UniversityState Key Laboratory of Bioreactor Engineering, East China University of Science and TechnologyAbstract Aspergillus oryzae is a widely used host for heterologous expression of fungal natural products. However, the vectors previously developed are not convenient for use and screening positive transformants by PCR and fermentation is time- and effort-consuming. Hence, three plug-and-play vectors were developed here for multi-gene expression and liquid chromatography mass spectrometry detection was introduced to screen positive transformants. Using rug BGC for verification, we demonstrated that the vectors we developed perform well and liquid chromatography mass spectrometry detection is feasible to screen positive transformants. For deleterious gene expression, PxyrA rather than PamyB was employed. Utilizing the toolkit described here to express natural products, dozen days can be saved. Graphical Abstracthttps://doi.org/10.1186/s40643-024-00833-wPlug-and-play vectorsLC–MS detectionAspergillus oryzaeHeterologous expressionPamyB |
| spellingShingle | Hanliang Shi Beibei Lin Mengmeng Zheng Fengyu Gan Zhi Lin Xiujuan Xin Jian Zhao Xudong Qu Faliang An Strategy of employing plug-and-play vectors and LC–MS screening to facilitate the discovery of natural products using Aspergillus oryzae Bioresources and Bioprocessing Plug-and-play vectors LC–MS detection Aspergillus oryzae Heterologous expression PamyB |
| title | Strategy of employing plug-and-play vectors and LC–MS screening to facilitate the discovery of natural products using Aspergillus oryzae |
| title_full | Strategy of employing plug-and-play vectors and LC–MS screening to facilitate the discovery of natural products using Aspergillus oryzae |
| title_fullStr | Strategy of employing plug-and-play vectors and LC–MS screening to facilitate the discovery of natural products using Aspergillus oryzae |
| title_full_unstemmed | Strategy of employing plug-and-play vectors and LC–MS screening to facilitate the discovery of natural products using Aspergillus oryzae |
| title_short | Strategy of employing plug-and-play vectors and LC–MS screening to facilitate the discovery of natural products using Aspergillus oryzae |
| title_sort | strategy of employing plug and play vectors and lc ms screening to facilitate the discovery of natural products using aspergillus oryzae |
| topic | Plug-and-play vectors LC–MS detection Aspergillus oryzae Heterologous expression PamyB |
| url | https://doi.org/10.1186/s40643-024-00833-w |
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