L-精氨酸生物合成的代谢流量分析.PDF

微生物学通报 MAR 20, 2008, 35(3) 395~401 Microbiology © 2008 by Institute of Microbiology, CAS tongbao@ 研究报告 L-精氨酸生物合成的代谢流量分析 朱进伟 陈青山 张伟国* 214122 摘 要: 建立并完善了谷氨酸棒杆菌 GWY020 及其 2 个逐步叠加不同遗传标记的突变株HUI821 和GUI089 合成L-精氨酸的中心代谢网络。分别测定了它们在特定培养时段(50 h~52 h)L-精氨酸 等代谢物的胞外浓度, 由此计算这一时段这些代谢物在发酵液中积累(或消耗) 的速率, 分别作出 这 3 株菌在拟稳态下的代谢流量分布图, 进而研究育种过程中不同遗传标记的叠加对代谢网络中 L-精氨酸合成流量分布的影响。结果表明遗传标记的引入使流量分配发生了重大变化, 节点处的 流量分配朝着有利于L-精氨酸合成的方向改变。从代谢流量分析角度上, 证明结构类似物抗性和 敏感性突变是代谢流导向和设计育种的有效手段, 代谢流量分析将成为设计育种的提供新思路。 关键词: L-精氨酸, 谷氨酸棒杆菌, 代谢流量分析 Metabolic Flux Analysis of L-Arginine Fermentation in Corynebacterium glutamicum * ZHU Jin-Wei CHEN Qing-Shan ZHANG Wei-Guo (The Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University , Wuxi 214122) Abstract: In this paper, metabolic networks of the Corynebacterium glutamicum GWY020 and the two de- rivatives carrying additional mutations HUI821and GUI089 were established and modified. The concentra- tions of extra-cellular metabolites were determined under sub-steady-state (50 h~52 h) of the batch culture. The metabolic flux distribution maps of the three strains were obtained, compared and analyzed. These re- sults indicate that the introduction of analog supersensitive marker or analog resistant marker skew the metabolic flux towards the formation of L-Arginine. This study revealed the usefulness