A newly isolated strain Enterococcus faecium INET2 was used as inoculum for biohydrogen production through dark fermentation. The individual and interactive effect of initial pH, operation temperature, glucose concentration and inoculation amount on the accumulation of hydrogen during fermentation was examined by a Box–Behnken Design (BBD), and hydrogen production process was analyzed at the optimal condition. A significant interactive effect between glucose concentration and pH was observed, the optimal condition was initial pH 7.1, operation temperature 34.8 °C, glucose concentration 11.3 g/L and inoculation amount 10.4%. Hydrogen yield, maximum hydrogen production rate and hydrogen production potential were determined to be 1.29 mol H₂/mol glucose, 86.7 L H₂/L/h and 1.35 L H₂/L. Metabolites analysis showed that E. faecium INET2 followed the pyruvate: formate lyase (Pfl) pathway in first 16 h, followed by the acetate-type fermentation and then shifted to butyrate-type fermentation. Maximum hydrogen production rate was accompanied with a quick formation of acetic acid.

新分离出的粪肠球菌INET2被用作接种菌，用于通过黑暗发酵生产生物氢。通过Box-Behnken设计（BBD）考察了初始pH，操作温度，葡萄糖浓度和接种量对发酵过程中氢积累的个体和交互作用，并在最佳条件下分析了氢的生产过程。观察到葡萄糖浓度与pH之间存在显着的相互作用，最佳条件为初始pH 7.1，操作温度34.8°C，葡萄糖浓度11.3 g / L和接种量10.4％。测得氢气产率，最大氢气产生速率和氢气产生潜能为1.29 mol H ₂ / mol葡萄糖，86.7 LH ₂/ L / h和1.35 LH ₂ / L。代谢物分析表明，粪肠球菌INET2在最初的16小时内遵循丙酮酸：甲酸裂解酶（Pfl）途径，随后进行乙酸盐型发酵，然后转变为丁酸盐型发酵。最大的氢气产生速率伴随着乙酸的快速形成。