Location: Renewable Product Technology ResearchTitle: Comparative investigation into formycin A and pyrazofurin A biosynthesis reveals branch pathways for the construction of C-nucleoside scaffolds
|ZHANG, MENG - Wuhan University|
|ZHANG, PEICHAO - Wuhan University|
|XU, GUDAN - Wuhan University|
|ZHOU, WENTING - Wuhan University|
|GAO, YAOJIE - Wuhan University|
|GONG, RONG - Wuhan University|
|CAI, YOU-SHENG - Wuhan University|
|CONG, HENGJIANG - Wuhan University|
|DENG, ZIXIN - Wuhan University|
|CHEN, WENQING - Wuhan University|
|MAO, XIANGZHOA - Ocean University Of China|
Submitted to: Applied and Environmental Microbiology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 8/7/2019
Publication Date: 8/7/2019
Citation: Zhang, M., Zhang, P., Xu, G., Zhou, W., Gao, Y., Gong, R., Cai, Y., Cong, H., Deng, Z., Price, N.P.J, Chen, W., Mao, X. 2019. Comparative investigation into formycin A and pyrazofurin A biosynthesis reveals branch pathways for the construction of C-nucleoside scaffolds. ACS Chemical Biology. https://doi.org/10.1101/728154.
Interpretive Summary: Resistance to commonly used antibiotics is a major threat to animal and human health, so continued development of new antimicrobials is a critical aspect of combating this problem. Formycin (FOR-A) and pyrazofurin (PRF-A) are antibiotics with potential agricultural applications that are produced by soil bacteria. Very little is known about how they are made, which has limited the ability to exploit the use of these unique antibiotics. In this work, we have identified the genes required for their biosynthesis and used this information to identify several intermediates in the metabolic pathway. This work not only provides information that can be used to overproduce and modify these antibiotics through genetic engineering, but it will also allow gene mining of other related antibiotics that may find uses as new antibiotics for agricultural and health purposes.
Technical Abstract: Formycin A (FOR-A) and pyrazofurin A (PRF-A) are purine-related C-nucleoside antibiotics in which ribose and a pyrazole-derived base are linked by a C-glycosidic bond. To date, the logic underlying the biosynthesis of these molecules has remained largely unexplored. Here, we report the discovery of the pathways for FOR-A and PRF-A biosynthesis from diverse actinobacteria, and demonstrate that the biosynthesis is initiated by a lysine N6-monooxygenase. Moreover, we show that forT and prfE mutants are correspondingly capable of accumulating unexpected pyrazole-related intermediates, compound 11 and 9a. Hence, PrfE/ForT is the first example of a ß-RFA-P (ß-ribofuranosyl-aminobenzene 5’-phosphate) synthase-like enzyme governing the construction of C-nucleoside scaffolds in natural products biosynthesis. We also, for the first time, decipher the enzymatic basis of the C-glycosidic bond formation in C-nucleoside antibiotics biosynthesis. The data establishes a foundation for combinational biosynthesis of related purine nucleoside antibiotics, and also open the way for target-directed genome mining of PRF-A/FOR-A related antibiotics.