(1) Development of metabolic reprogramming technologies for methylotrophic microorganisms

Addressing challenges such as the low efficiency of exogenous DNA transformation in methylotrophic microorganisms, the absence of effective genetic modification and metabolic regulation technologies, and limitations in both foundational and applied research, this project systematically investigates the DNA defense and repair mechanisms in methylotrophic microorganisms. Its goal is to reprogram these defense and repair networks to improve exogenous DNA transformation efficiency. Additionally, the project develops advanced gene editing and regulatory tools based on CRISPR and base modification enzymes, providing technical support for exploring microbial C1 metabolism regulatory mechanisms and creating efficient C1 cell factories.

(2) Investigation of regulatory mechanisms in microbial C1 metabolism

Focusing on natural methylotrophic microorganisms and synthetic methylotrophic Corynebacterium glutamicum strains developed in previous studies, this research investigates the metabolic regulatory differences in microbial cells when utilizing C1 versus multi-carbon feedstocks. The project aims to map the metabolic pathways and regulatory networks governing C1 metabolism and to identify key genetic targets that influence these pathways. By integrating synthetic and systems biology approaches, it seeks to clarify the mechanisms of tolerance, metabolism, and regulation of C1 compounds in microorganisms, thereby providing a theoretical foundation for efficient methanol bioconversion.

(3) Design and creation of C1 cell factories

Building on insights into C1 metabolism regulation, this research enhances the tolerance and utilization capacities of both natural and synthetic methylotrophic microorganisms for C1 compounds through metabolic reprogramming, facilitating the creation of novel C1 chassis cells. With a focus on bio-based material monomers, amino acids, other small biomolecules, and single-cell proteins, the project aims to design synthetic pathways with high atom economy and energy efficiency to convert C1 molecules into multi-carbon products. These pathways are then assembled and optimized in the new chassis cells to enable the efficient production of multi-carbon products from methanol and other C1 feedstocks.