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细履平沙

Abstract The recent progress on metabolic systems engineering was reviewed, in particular focusing on the metabolic flux analysis (MFA) based on the isotopomer distribution obtained using NMR and/or GC-MS.After the brief explanation of how to estimate the metabolic flux distribution (MFD) based on 13C-labeling experiments, the metabolic regulation analysis was made based on the protein expression patterns obtained by two-dimensional electrophoresis (2DE) together with enzyme activity data, and the gene expression patterns obtained by RT-PCR analysis.The particular application was considered for Escherichia coli.
The effect of culture conditions such as different carbon sources (glucose, gluconate, glycerol, acetate, etc.) and different dissolved oxygen (DO) concentration, etc. on the metabolism was investigated.The effect of some single-gene knockout such as pgi–, pyk–, and gnd–was also investigated. It was found to be quite useful to integrate the information obtained from metabolic flux analysis, gene and protein expressions as well as intracellular metabolite concentrations to understand the overall picture of metabolic regulation.
摘要:本文综述了最近几年在代谢系统工程方面的发展,特别关注了基于同位素分布的代谢流分析,同位素分布可以通过核磁和/或气质联用检测.在简要介绍了如何通过C13标记实验对代谢流分布进行估计后,通过2维电泳和酶活性数据得出了蛋白表达模式；通过RT-PCR分析得到了基因表达模式,基于这两个模式进行了代谢调控分析。在大肠杆菌中进行了应用。
培养条件如不同的C源（葡萄糖，葡萄糖酸盐，甘油，乙酸等）和不同的DO浓度等对代谢的影响进行了研究。一些单基因敲除如pgi-,pyk-t gnd-也进行了研究。研究发现将来自代谢流分析，基因和蛋白表达的信息及细胞内代谢浓度进行综合对于理解代谢调控的全局很有用处.