Abstract:The development of green and low-carbon renewable energy systems has become an important international consensus. It is also an essential path for China to implement the dual-carbon strategy, ensure national energy security, and achieve sustainable development. This review introduces the theory of a new energy system based on electricity-hydrogen-carbohydrate (EHC) cycle, and highlights the biotransformations of carbohydrate/water-to-hydrogen, carbohydrate-to-electricity, and CO2-to-carbohydrate powered by hydrogen- or electric-energy based on the in vitro synthetic enzymatic biosystems (ivSEB) developed by Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences in the past decade. We elaborate the design principle and the molecular basis of ivSEB, and further expand from the EHC cycle to in vitro biomanufacturing with starch as the feedstock. Combined with the latest research advances, we analyze and discuss advantages and disadvantages of ivSEB, prospect future directions, so as to promote the green, low-carbon and sustainable development of economy and society.