Intelligent Design of Microbial Carbohydrate Factories: Integrated Chassis-Tools-Strategies Frameworks.

Carbohydrates, natural polymers of monosaccharides interconnected via glycosidic bonds, serve as versatile biomolecules with broad applications across food, pharmaceuticals, agriculture, and chemical industries. Traditionally, carbohydrate production has predominantly relied on extraction or chemical synthesis methods; however, with the progression of synthetic biology, microbial production of carbohydrates has emerged as a promising alternative, offering rapid and environmentally sustainable manufacturing processes. This review presents integrated methodologies for engineering carbohydrate-synthesizing microbial factories. We first elucidate the fundamental physiological and metabolic characteristics of industrial chassis organisms, followed by systematic evaluation of computational and experimental tools for predictive modeling and precise modulation of carbohydrate anabolic pathways. Building on this foundation, we critically analyze cutting-edge metabolic engineering strategies spanning pathway optimization, pathway reconstruction, and dynamic control mechanisms, thereby establishing a multidimensional theoretical framework for intelligent design of carbohydrate biosynthesis systems.