酮基还原酶基因工程菌发酵特性的研究  

Fermentation performance of genetically engineered ketoreductase-producing bacteria

作　　者：张松[1] 李啸[1,2] 石小丹[2] 程奔[1] 

机构地区：[1]三峡大学生物与制药学院,湖北宜昌443003 [2]安琪酵母股份有限公司,湖北宜昌443003

出　　处：《中国酿造》2015年第7期69-73,共5页China Brewing

基　　金：湖北省重大科技专项项目(2012ACA15)

摘　　要：主要研究了基因工程大肠杆菌发酵生产酮基还原酶的发酵特性。结果表明，发酵12h菌体浓度达到最大，发酵12-22h菌体浓度稳定；发酵液pH值先平稳后急速上升至pH值8．6左右，之后趋于平稳；6～10h达到碳氮源利用高峰，14h后氨基酸态氮含量略微回升；乙酸于10h质量浓度最大，10～12h迅速下降后趋于平稳；酶活于12h达到最大值92．9U／mL，之后，利用流加氨苄的对照罐发酵，验证了发酵特性曲线的可靠性，菌体质粒稳定性得到提升，最大酶活优化为125．2U／mL。此研究对于产酮基还原酶的基因工程菌进一步的发酵优化具有指导意义。The main subject of this topic was to study the fermentation characteristics of recombinant Escherichia coli for ketoreductase production. The results showed that the cell concentration was the maximum at 12 h, and it was steady from 12-22 h. The pH of the fermentation broth was stable first, and then increased rapidly to approximately 8.6 at 10-12 h, finally leveled off. At 6-10 h, the utilization of carbon and nitrogen source reached the peak, and after 14 h, the amino acid nitrogen content slightly rose again. The acetic acid concentration was measured to the maximum at 10 h, and little changed after the acetic acid concentration decreased rapidly from 10 to 12 h; Ketoreductase activity was examined to the maximum of 92.9 U/ml at 12 h. Then, by the fermentor supplemented with ampicillin as control, the reliability of the fermentation characteristics was verified, and the bacte- rial plasmid stability was enhanced, the maximum activity was optimized to 125.2 U/ml. The study could provide a reference for further optimization of gene engineering ketoreductase-producing strain fermentation.

关 键 词：酮基还原酶 发酵特性 酶活 质粒稳定性 

分 类 号：O657.32[理学—分析化学]