Physiological and metabolic analysis of nitrate reduction on poly-gamma-glutamic acid synthesis in Bacillus licheniformis WX-02

Nitrate is an important nitrogen source for organism, but whether and how nitrate improves poly-γ-glutamic acid (γ-PGA) production of bacterial is not clear. The effect of nitrate on γ-PGA production of Bacillus licheniformis WX-02 was investigated. By addition of 50 mmol/L nitrate, the γ-PGA yield reached 12.3 ± 0.21 g/L, which increased 2.3-fold compared to the control. The mechanism of enhanced γ-PGA production was further investigated by analysis of nitrate reduction, physiology, pyruvate overflow metabolism and energy synthesis. Nitrate reduction was only carried out in the middle stage of γ-PGA fermentation. The result of consumption of nutrients showed that glucose uptake was not effected and the l-glutamic acid utilization efficiency increased from 48.3 to 77.0 %. The date of overflow metabolism obtained from high-performance liquid chromatography showed that the metabolism of pyruvate, formate, lactate and acetoin was both heightened by nitrate reduction, while the 2,3-butanediol biosynthesis was decreased. Meanwhile, the change of energy indicated that more ATP was synthesized during nitrate reduction. In summary, nitrate was a positive effector of γ-PGA biosynthesis in B. licheniformis WX-02 and nitrate reduction affected multi-metabolism pathways, including glycolysis, overflow metabolism and energy metabolism.