代谢组学分析提高谷氨酸棒杆菌L-瓜氨酸产量  

Improve L-citrulline production in Corynebacterium glutamicum by metabolomics analysis

作　　者：许娟 许琳[1] 郝宁[1] XU Juan;XU Lin;HAO Ning(College of Biotechnology and Pharmaceutical Engineering,Nanjing Tech University,Nanjing 211800,China)

机构地区：[1]南京工业大学生物与制药工程学院,江苏南京211800

出　　处：《生物加工过程》2018年第4期8-15,共8页Chinese Journal of Bioprocess Engineering

基　　金：国家高技术研究发展计划(863计划)(2012AA022101);国家自然科学基金(21406113);江苏省自然科学基金(BK20140932)

摘　　要：利用基于气相色谱-质谱联用(GC-MS)的代谢组学分析方法,研究Corynebacterium glutamicum ATCC 13032和C.glutamicum ATCC 13032Δarg GΔargR-p XMJ19-arg B(ec)(CIT4)重组菌胞内代谢物差异性,挖掘L-瓜氨酸合成关键代谢物及代谢途径作用机制,并通过添加关键代谢物进行理性强化,提高L-瓜氨酸的产量。利用GC-MS共检测124种化合物,结合主成分分析(PCA)和偏最小二乘判别分析(PLS)统计发现10种为区别这两种菌的生物标志物,理性强化添加α-酮戊二酸、丙酮酸、谷氨酸、鸟氨酸、氨甲酰磷酸和谷氨酰胺,CIT4中L-瓜氨酸产量从18.50 g/L增加到20.86 g/L。The intracellular metabolic differences of Corynebacterium glutamicum ATCC 13032 and C.glutamicum ATCC 13032Δarg GΔargR-p XMJ19-arg B（ ec）（ CIT4） recombinant bacteria were studied by metabolomics analysis based on gas chromatography-mass spectrometry（ GC-MS）.In this way,we studied the differences between them,to learn how the key metabolites worked in metabolic pathway of L-citrulline synthesis,and to improve the yield of L-citrulline by adding key metabolites for rational strengthening. A total of 124 compounds were detected by GC-MS,and 10 biomarkers were identified to be mainly difference between these two strains by principal component analysis（ PCA） and partial least squares（ PLS）. Through the exogenous addition strategies of adding α-ketoglutaric acid,pyruvate,glutamic acid,ornithine,carbamoyl phosphate and glutamine,the yield of L-citrulline in CIT4 increased from 18. 50 g/L to 20. 86 g/L.

关 键 词：谷氨酸棒杆菌 L-瓜氨酸 代谢组学 理性培养基强化 

分 类 号：TQ922[轻工技术与工程—发酵工程]