From manufacturing to intelligent manufacturing: the symbiotic tale of synthetic biology and vitamin B<sub>12</sub>

<p indent="0mm">Synthetic biology, which approaches biotechnological problems with an engineering mindset, integrates knowledge from disciplines such as physics, chemistry, and computer science, aiming to transform, design, and construct novel biological systems. This interdisciplinary field is distinct in its systematic, modular, and design-oriented methodology, which contrasts with the more traditional trial-and-error techniques of biology. By applying engineering principles, synthetic biology enables precise manipulation and optimization of biological processes, leading to significant advancements in biotechnology. One particularly valuable compound that has benefited from this approach is vitamin B₁₂. Vitamin B₁₂ is a valuable compound with diverse applications in medicine, chemistry, and biology, which has inspired continuous research for nearly a century. Early breakthroughs in vitamin B₁₂ research, especially those related to its chemical structure and biological functions, earned several Nobel Prizes and contributed to fundamental knowledge in medical and chemical fields. Currently, the production of vitamin B₁₂ has traditionally relied on natural microbial strains, which synthesize the compound through complex biochemical pathways. Recently, its biotechnological production has received a fresh impetus due to advances of synthetic biology. This paper explores the important achievements of vitamin B₁₂ research since its discovery in 1926, including the Nobel prize-winning achievements during the early periods of medical and chemical investigation, which laid the groundwork for understanding the compound’s structure and function. Additionally, since the 1980s, the isolation of naturally occurring strains that synthesize vitamin B₁₂, along with the analysis of their catalytic pathways, has sparked a surge of interest in the biotechnological production of vitamin B₁₂. Subsequently, the development of genetic tools of natural strains capable of synthesizing vitamin B₁₂ has significantly advanced its biosynthetic production. Recently, construction and optimization of engineered vitamin B₁₂ production strains using <italic>Escherichia coli</italic>, along with synthetic systems based on cell-free multienzyme catalysis, have been reported. These advancements highlight promising new strategies for the future of biotechnological vitamin B₁₂ synthesis. In summary, this article particularly highlights the significant promotion of vitamin B₁₂ biomanufacturing by the introduction and advancement of systematic synthetic biology research methods in the past decade. These advancements have propelled vitamin B₁₂ production from a simple “manufacturing” process based on natural strains towards a more sophisticated, intelligent, technology-driven approach. By introducing systematic synthetic biology methods, such as developing efficient genetic engineering technologies for natural synthesis strains, creating artificial strains capable of producing vitamin B₁₂, and assembling flexible cell-free vitamin B₁₂ synthesis systems—The production process has become not only more efficient but also highly controllable, enabling precise regulation of biosynthesis. The integration of computational tools, systems biology, and high-throughput screening has further accelerated the optimization of production systems, making the process more intelligent and design-driven. Furthermore, the intricate natural synthesis pathway of vitamin B₁₂ serves as a paradigm for synthetic biology research. Its complexity and the challenges associated with its synthesis offer an excellent classical example of how synthetic biology can be applied to solve intricate biological problems. By studying and modifying this pathway, researchers can better understand the principles of pathway engineering and apply these insights to other complex biosynthetic processes. In doing so, synthetic biology continues to drive the evolution of vitamin B₁₂ production and other biotechnological applications, promoting innovation and furthering the development of the field.