Elucidation of the complete biosynthetic pathway of the main triterpene glycosylation products of Panax notoginseng using a synthetic biology platform |
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| Institution: | 1. Tianjin Key Laboratory of Industrial Microbiology, College of Biotechnology, Tianjin University of Science and Technology, Tianjin 300457, China;2. State Key Laboratory of Biocatalysis and Enzyme Engineering, Hubei Collaborative Innovation Center for Green Transformation of Bio-Resources, Hubei Key Laboratory of Industrial Biotechnology, School of Life Sciences, Hubei University, Wuhan 430062, China;1. Department of Plant Biotechnology and Bioinformatics, Ghent University, 9052 Ghent, Belgium;2. Center for Plant Systems Biology, VIB, 9052 Ghent, Belgium;3. Laboratory for Protein Biochemistry and Biomolecular Engineering, Department of Biochemistry and Microbiology, Ghent University, B-9000 Ghent, Belgium;4. VIB Medical Biotechnology Center, B-9000 Ghent, Belgium;1. Institute for Synthetic Biosystem, Department of Biochemical Engineering, School of Chemistry and Chemical Engineering, Beijing Institute of Technology, Beijing 100081, China;2. Key Laboratory of Systems Bioengineering (Ministry of Education), School of Chemical Engineering and Technology, Tianjin University, Tianjin 300072, China;1. School of Biological and Agricultural Engineering, Jilin University, No.5988, Renmin Street, Nanguan District, Changchun, PR China;2. College of Life Sciences, Jilin University, No.2699, Qianjin Street, Chaoyang District, Changchun, PR China;1. State Key Laboratory of Microbial Metabolism, School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai 200240, China;2. Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin 300308, China;1. Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, China;2. Key Laboratory of Systems Microbial Biotechnology, Chinese Academy of Sciences, China;3. College of Biotechnology, Tianjin University of Science and Technology, China;4. National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences, China;5. School of Traditional Chinese Medicine, Capital Medical University, China |
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| Abstract: | UDP-glycosyltransferase (UGT)-mediated glycosylation is a widespread modification of plant natural products (PNPs), which exhibit a wide range of bioactivities, and are of great pharmaceutical, ecological and agricultural significance. However, functional annotation is available for less than 2% of the family 1 UGTs, which currently has 20,000 members that are known to glycosylate several classes of PNPs. This low percentage illustrates the difficulty of experimental study and accurate prediction of their function. Here, a synthetic biology platform for elucidating the UGT-mediated glycosylation process of PNPs was established, including glycosyltransferases dependent on UDP-glucose and UDP-xylose. This platform is based on reconstructing the specific PNPs biosynthetic pathways in dedicated microbial yeast chassis by the simple method of plug-and-play. Five UGT enzymes were identified as responsible for the biosynthesis of the main glycosylation products of triterpenes in Panax notoginseng, including a novel UDP-xylose dependent glycosyltransferase enzyme for notoginsenoside R1 biosynthesis. Additionally, we constructed a yeast cell factory that yields >1 g/L of ginsenoside compound K. This platform for functional gene identification and strain engineering can serve as the basis for creating alternative sources of important natural products and thereby protecting natural plant resources. |
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| Keywords: | UDP-glycosyltransferases Notoginsenoside R1 Yeast cell factories Synthetic biology |
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