Riboflavin synthesis from gaseous nitrogen and carbon dioxide by a hybrid inorganic-biological system

Microbes can provide a more sustainable and energy-efficient method of food and nutrient production compared to plant and animal sources, but energy-intensive carbon (e.g., sugars) and nitrogen (e.g., ammonia) inputs are required. Gas-fixing microorganisms that can grow on H(2) from renewable water splitting and gaseous CO(2) and N(2) offer a renewable path to overcoming these limitations but confront challenges owing to the scarcity of genetic engineering in such organisms. Here, we demonstrate that the hydrogen-oxidizing carbon- and nitrogen-fixing microorganism Xanthobacter autotrophicus grown on a CO(2)/N(2)/H(2) gas mixture can overproduce the vitamin riboflavin (vitamin B(2)). We identify plasmids and promoters for use in this bacterium and employ a constitutive promoter to overexpress riboflavin pathway enzymes. Riboflavin production is quantified at 15 times that of the wild-type organism. We demonstrate that riboflavin overproduction is maintained when the bacterium is grown under hybrid inorganic-biological conditions, in which H(2) from water splitting, along with CO(2) and N(2), is fed to the bacterium, establishing the viability of the approach to sustainably produce food and nutrients.