Engineering a living cell to desired metabolite concentrations and fluxes: pathways with multifunctional enzymes.
Engineering a living cell to desired metabolite concentrations and fluxes: pathways with multifunctional enzymes.
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将活细胞改造为所需的代谢物浓度和通量：多功能酶的途径。
DOI：
10.1006/mben.1999.0132
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发表时间：
2000
影响因子：
8.4
通讯作者：
Cascante,M
中科院分区：
文献类型：
--
作者：
Kholodenko,BN;Westerhoff,HV;Schwaber,J;Cascante,M
由于分子遗传学能够调节细胞酶的浓度，代谢工程受到这样的问题的限制，即需要对酶浓度进行哪些调节才能产生所需的细胞代谢模式。在较早的一篇论文中(Kholodenko等人，(1998年)。生物技术。比昂斯。59,239-247)，我们推导出一种确定所需调制的方法。然而，这种方法不能立即应用于酶催化新陈代谢的一个以上步骤(多功能酶)的细胞途径。在本文中，我们展示了多功能酶的存在在多大程度上限制了生物技术的雄心，否则人们可能会徒劳地追求这些雄心。特别是，如果中间体直接与多功能酶相互作用，就不可能改变单一中间体的浓度，而不会影响代谢的其余部分。新陈代谢控制分析的分析机制被用来联系所需的和随后的代谢模式的变化。然后，以单矩阵方程的形式给出了工程新陈代谢问题的显式解。
With molecular genetics enabling modulation of the concentrations of cellular enzymes, metabolic engineering becomes limited by the question of which modulations of the enzyme concentrations are required to bring about a desired pattern of cellular metabolism. In an earlier paper (Kholodenko et al. (1998). Biotechnol. Bioeng. 59, 239–247) we derived a method to determine the required modulations. This method, however, cannot be immediately applied to cellular pathways with enzymes catalyzing more than one step in metabolism (multifunctional enzymes). In the present paper we show to which extent the presence of multifunctional enzymes limits biotechological ambitions, which one might otherwise pursue in vain. In particular, it is impossible to change the concentration of a single intermediate and leave the rest of metabolism unperturbed if that intermediate interacts directly with a multifunctional enzyme. The analytical machinery of Metabolic Control Analysis is used to relate the desired and ensuing changes in the metabolic pattern. An explicit solution to this problem of engineering metabolism is then given in the form of a single matrix equation.