Mapping and Redirecting Metabolic Fluxes to Attain Significantly Lessen Oxygen Requirements, Acid Production, and Enhanced Recombinant Culture Productivity

映射和重定向代谢通量，以显着减少需氧量、酸产生并增强重组培养生产力

• 批准号: 9622821
• 负责人: Mohammad Ataai
• 金额: $ 46.09万
• 依托单位: University of Pittsburgh
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 1996
• 资助国家: 美国
• 起止时间: 1996-08-15 至 2000-01-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=9622821&HistoricalAwards=false
• 关键词: Mapping; Redirecting; Metabolic; Fluxes; Attain

9622821 Ataai This proposal is on research which addresses one of the bottlenecks in protein production frequently reported for recombinant E. coli, namely overflow production of semi-oxidized sugars and carbohydrates (e.g. acetate) which reduces the total carbon yield and also indicates inefficient metabolic biosynthetic/catabolic fluxes. The specific goals of this research are to: (1) build on the investigators' Bacillus research to experimentally validate (using NMR) the feasibility of optimized flux distribution centered around pyruvate kinase (PYK), (2) transfer the yield-increasing strategy to E. coli by deleting PYK isozymes and analyze via metabolic flux analysis, again coupled with NMR, (3) determine scale-up potential in terms of mutant E. coli concentration, recombinant protein production, and oxygen consumption, and, (4) evaluate whether there is a link between energy-consuming proteolysis and ATP-generation as shown in high glucose feed E. coli cultures. ***

9622821 Ataai该建议是关于解决重组大肠杆菌蛋白质生产中经常报道的瓶颈之一的研究。大肠杆菌中，即半氧化糖和碳水化合物（例如乙酸盐）的溢流产生，这降低了总碳产率，并且还指示低效的代谢生物合成/分解代谢通量。 本研究的具体目标是：（1）在芽孢杆菌研究的基础上，利用核磁共振技术验证以丙酮酸激酶（PYK）为中心的优化流量分布的可行性;通过删除PYK同工酶并通过代谢通量分析进行分析，再次与NMR结合，（3）确定突变体E.大肠杆菌浓度、重组蛋白产量和耗氧量，以及（4）评估高葡萄糖饲料中所示的耗能蛋白水解和ATP生成之间是否存在联系。大肠杆菌培养物。 ***