组合添加代谢调控剂对肌苷发酵及其代谢流分布的影响  

Effects of Assembling Metabolic Regulation Agents on Inosine Fermentation and Metabolic Flux Distributions

作　　者：肖安风[1,2,3] 杨尚彤 倪辉[1,2,3] 杨哲[1] 蔡慧农 

机构地区：[1]集美大学生物工程学院,福建厦门361021 [2]福建省食品微生物与酶工程重点实验室,福建厦门361021 [3]厦门市食品生物工程技术研究中心,福建厦门361021

出　　处：《高校化学工程学报》2014年第4期791-797,共7页Journal of Chemical Engineering of Chinese Universities

基　　金：福建省产学研重大专项项目(2010N5009);福建省自然科学基金项目(2011J01224);集美大学中青年创新团队专项基金项目(2010A006)资助

摘　　要：根据肌苷合成的代谢调控机理,选择五种对肌苷合成有促进作用的代谢调控剂—氟化钠、次黄嘌呤、葡萄糖酸钙、氯化钙和柠檬酸钠,并确定了其最适添加量。进而组合添加这些代谢调控剂,对肌苷生物合成过程的多个关键代谢调控节点进行调控,增大流向肌苷合成的代谢流,使肌苷产量得到提高。研究发现,在培养基中组合添加这些代谢调控剂,所得肌苷产量比单一添加代谢调控剂时的肌苷产量有显著提升。其中当组合添加2 mg·L-1 NaF、0.15%次黄嘌呤、15 g·L-1葡萄糖酸钙及10 mg·L-1 CaCl2时,肌苷产量达到了14.01 g·L-1,相比对照组的肌苷产量提高了3.39倍,协同效应显著。继而,对肌苷合成的代谢网络各分支途径的代谢通量进行了测定,证实了组合添加代谢调控剂有利于葡萄糖代谢流由EMP途径向HMP途径发生迁移,实现对葡萄糖主体代谢流分布的调整,促使肌苷合成代谢流增加,从而有利于肌苷产量的进一步提升。Following the regulating mechanism in inosine synthesis, sodium fluoride, hypoxanthine, calcium gluconate, calcium chloride and sodium citrate were used as additives for their positive effects on inosine production, and the amounts used were optimized. The key nodes in the inosine biosynthetic process were studied, and the metabolism flux was increased to promote the inosine production. The results show that the combination of these additives can increase the inosine yield which is better than that of a single additive. A combination of 0.15% hypoxanthine, 2 mg.L^-1 sodium fluoride, 10 mg.L^-1 calcium chloride and 15 g.L^-1 calcium gluconate shows an output of 14.01 g.L ^-1, which is about 3.39 times higher than using any single additive. Meanwhile, measurements of the branch synthesis metabolic pathway shows that the combination of these additives does help the glucose metabolism flux moving from EMP pathway to HMP pathway, thus increases inosine biosynthesis production.

关 键 词：枯草芽孢杆菌 肌苷 发酵过程 代谢调控剂 代谢通量分析 

分 类 号：Q815[生物学—生物工程]