2,3-Butanediol (2,3-BD) is an important platform chemical that can be produced biologically. It is currently fermented below 40 °C using mesophilic strains at acid to neutral pH conditions, but the processes often suffer bacterial contamination. Here we reported a new nonpathogenic strain Bacillus licheniformis YNP5-TSU isolated from Yellowstone National Park, which produces 2,3-BD with high yield and productivity at thermophilic and alkaline conditions. B. licheniformis YNP5-TSU has the ability to survive in pH ranges from 8.0 to 10.0 and temperature ranges from 40 to 55 °C, which greatly reduces the chance of contamination from other microorganisms. At its optimum condition with a temperature of 50 °C and pH of 8–9, YNP5-TSU can produce 27.4 g/L 2,3-BD from 60 g/L glucose, with a high yield of 0.46 g/g (92% of theoretical value) and productivity of 1.1 g/L/h in a batch fermentation. YNP5-TSU also can ferment different carbohydrates including fructose, sucrose, arabinose, galactose, mannose, xylose, as well as alcohols such as glycerol. The effects of nitrogen and mineral sources on 2,3-BD fermentation were also investigated to improve the fermentation process. In a fed-batch fermentation, 99.3 g/L of 2,3-BD was obtained within 96 h, with a yield of 0.41 g/g. The results provided indicate that B. licheniformis YNP5-TSU is a very adaptable strain to withstand harsh environments while still producing 2,3-BD near theoretical values.