Rubisco-Centric Strategies for Carbon Conservation in Synthetic Biology

The escalating global climate crisis urgently demands biomanufacturing technologies with higher carbon efficiency. Ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco), the central enzyme catalyzing carbon dioxide fixation in the Calvin-Benson cycle, exhibits low efficiency, thereby limiting its industrial application. This review summarizes fundamental Rubisco research, laying the foundation for its efficiency-enhancing engineering and biomanufacturing applications. We first outline the discovery, systematic classification, and molecular evolution of Rubisco, and then elucidate the structural dynamics underlying its activation and inhibition mechanisms. Subsequently, we provide the first systematic synthesis of five enzyme-efficiency enhancement strategies, spanning classical directed evolution to artificial intelligence-driven rational design. Recent advances in the utilization of Rubisco by autotrophic and heterotrophic microorganisms for the synthesis of biobased products are also critically examined. The resulting cross-scale design framework integrates evolutionary biology, structural dynamics, and synthetic biology, offering an extensible paradigm for sustainable, carbon-neutral biomanufacturing.