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过程工程学报 ›› 2017, Vol. 17 ›› Issue (4): 814-820.DOI: 10.12034/j.issn.1009-606X.216335

• 生化工程 • 上一篇    下一篇

不透明红球菌生产谷氨酸氧化酶发酵过程优化

刘 佳1, 徐继嗣2, 罗秋玲1, 陈修来1, 刘立明1*   

  1. 1. 江南大学食品科学与技术国家重点实验室,工业生物技术教育部重点实验室,江苏 无锡 214122; 2. 张家港市华天制药有限公司,江苏 张家港 215633
  • 收稿日期:2016-10-26 修回日期:2016-12-28 出版日期:2017-08-20 发布日期:2017-08-16
  • 通讯作者: 刘立明 mingll@jiangnan.edu.cn
  • 基金资助:
    国家自然科学基金资助项目;江苏省科技支撑计划社会发展项目

Optimization of fermentation process for production of glutamic acid oxidase from Rhodococcus opacus

Jia LIU1,  Jisi XU2,  Qiuling LUO1,  Xiulai CHEN1,  Liming LIU1*   

  1. 1. State Key Lab. Food Sci. Technol., Key Lab. Ind. Biotechnol., Ministry of Education, Jiangnan Univ., Wuxi, Jiangsu 214122, China; 
    2. Huatian Pharmaceutical Co., Ltd., Zhangjiagang, Jiangsu 215633, China
  • Received:2016-10-26 Revised:2016-12-28 Online:2017-08-20 Published:2017-08-16
  • Contact: LI Zhen mingll@jiangnan.edu.cn

摘要: 对自主筛选的不透明红球菌FMME1-41进行发酵营养条件优化,得到最佳发酵培养基:酵母粉6 g/L,大豆蛋白胨2 g/L,(NH4)2SO4 0.8 g/L,葡萄糖25 g/L,KH2PO4 3 g/L,MgSO4 0.6 g/L,MnSO4 0.3 g/L,L-谷氨酸2.5 g/L,在最佳培养基中发酵30 h,LOGX酶活达到6.12 U/mL.在7.5 L发酵罐中进行补料策略优化,使用分批发酵和DO-stat补料相结合的两阶段发酵方法,发酵周期40 h,LOGX酶活达到21.5 U/mL,是分批发酵LOGX酶活的2.8倍. 利用LOGX在2 L体系中转化L-谷氨酸生产α-酮戊二酸(α-KG),α-KG产量达到92.0 g/L,摩尔转化率为92.6%,α-KG生产强度为9.2 g/L/h.

关键词: 不透明红球菌, 谷氨酸氧化酶, 发酵优化, α-酮戊二酸

Abstract: The optimization of fermentation conditions for the independent screening strain Rhodococcus opacus FMME1-41 was carried out. The optimal nutrition conditions was as follows: 6 g/L yeast powder, 2 g/L soya peptone, 0.8 g/L (NH4)2SO4, 25 g/L glucose, 3 g/L KH2PO4, 0.6 g/L MgSO4, 0.3 g/L MnSO4, 2.5 g/L glutamic acid. In addition, fermentation period was 30 h, the LOGX activity reached 6.12 U/mL. Feeding strategy was optimized in 7.5 L fermentation tank. Under the condition of combination of bat. ch fermentation and DO-stat feeding, the fermentation period was 40 h, and the LOGX activity was 21.5 U/mL, which is 2.8 times of the batch fermentation. LGOX was used to produce α-ketoglutaric acid (α-KG) from L-glutamic acid in 2 L systerm. Finally, the maximum titer of α-KG reached 92 g/L, the molar conversion rate was 92.6%, the production intensity of α-KG was 9.2 g/L/h.

Key words: Key words: Rhodococcus opacus, L-Glutamate Oxidase, fermentation optimization, α-Ketoglutaric Acid