为提高谷氨酸棒杆菌发酵产L-谷氨酰胺产量、糖酸转化率、生产强度等技术指标,该文通过响应面法优化了摇瓶发酵培养基中葡萄糖、(NH4)2SO4和玉米浆含量,并利用50 L发酵罐进行分批发酵,构建了菌体生长、底物消耗和产物生成动力学模型,进而在50 L发酵罐中采用两阶段控制pH策略,并通过补加葡萄糖和(NH4)2SO4,协同控制发酵液中碳氮源浓度,促进菌体高效合成L-谷氨酰胺。结果表明,优化后发酵培养基中葡萄糖、(NH4)2SO4和玉米浆含量分别为167、61、31 g/L,摇瓶发酵L-谷氨酰胺产量达到38.13 g/L,较优化前提高了34.12%;50 L发酵罐分批发酵80 h后L-谷氨酰胺产量达42.06 g/L,糖酸转化率达到25.85%,生产强度为0.53 g/(L·h);在50 L发酵罐中基于两阶段pH控制的碳氮源协同补加策略,显著促进了谷氨酸棒杆菌高产L-谷氨酰胺,发酵周期缩短至52 h, L-谷氨酰胺产量达68.01 g/L,糖酸转化率达33.93%,生产强度为1.31 g/(L·h)。该研究对促进L-谷氨酰胺的工业化生产具有重要参考价值。
To improve the fermentation level, sugar-acid conversion rate, and productivity of L-glutamine production by Corynebacterium glutamicum, the contents of glucose, ammonium sulfate, and corn syrup in fermentation medium were optimized using response surface methodology. Then the kinetic models for cell growth, substrate consumption and L-glutamine production of batch fermentation in 50 L fermenter were constructed. Furthermore, the strategy of two-stage pH control combined with coordinating the concentrations of carbon and nitrogen sources by adding glucose and ammonium sulfate in 50 L fermenter was adopted to promote L-glutamine production. The results showed that the optimized fermentation medium contained 167 g/L glucose, 61 g/L (NH4)2SO4, and 31 g/L corn syrup, respectively. L-Glutamine yield in shaking flask fermentation reached 38.13 g/L, an increase of 34.12% compared with that before optimization. After 80 h of batch fermentation in 50 L fermenter, L-glutamine yield reached 42.06 g/L, the sugar-acid conversion rate reached 25.85%, and the productivity was 0.53 g/(L·h). The strategy of two-stage pH control combined with synergistic addition of carbon and nitrogen sources in 50 L fermenter significantly promoted L-glutamine production, shortening the fermentation period to 52 h, achieving 68.01 g/L of L-glutamine, 33.93% of sugar-acid conversion and 1.31 g/(L·h) of productivity. This study provides valuable guidance for promoting industrial production of L-glutamine by C. glutamicum.
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