The Technology Research Association for Next-Generation Natural Products Chemistry (N2PC) has been developing cutting-edge technologies for cloning and heterologous expression of biosynthesis gene clusters, which are sometimes large DNA fragments exceeding 100kb with high homology and repeated sequences, for industrially useful natural compounds. In this project, N2PC is working on development of means to modify the core structure of medium-sized-molecule natural products by applying these technologies.
- Development of mother nucleus modification technology for natural compounds
N2PC succeeded in developing technology using BAC clones in vitro that enables accurate genome editing at targeted positions, instead of conventional homologous recombination technology performed inside cells (in vitro module editing technology). By advancing the in vitro module editing technology developed, N2PC has also succeeded, for the first time in the world, in developing technology to precisely modify the biosynthetic gene module of medium-sized-molecule natural compounds, which usually have extremely high homology and repeat sequences.
- Development of conversion technology for medium-sized molecules using microbial enzymes
A large number of cytochrome P450 enzymes were collected for the purpose of metabolite analysis of medium-sized-molecule natural compounds. In addition, we have developed a cloning vector system for the optimization of P450 enzyme activity, and also succeeded in developing an enzyme activity expression system that abolished the production of by-products and significantly improved the maximum activity. In addition, we have succeeded in developing technology that can efficiently identify the target enzyme from among many candidate enzymes present in a microorganism.