Institute of Bio- and Geosciences, Biotechnology, Systems Biotechnology, Forschungszentrum Jülich GmbH, Jülich, Germany.
Microb Cell Fact. 2012 Sep 11;11:122. doi: 10.1186/1475-2859-11-122.
Overflow metabolism is well known for yeast, bacteria and mammalian cells. It typically occurs under glucose excess conditions and is characterized by excretions of by-products such as ethanol, acetate or lactate. This phenomenon, also denoted the short-term Crabtree effect, has been extensively studied over the past few decades, however, its basic regulatory mechanism and functional role in metabolism is still unknown. Here we present a comprehensive quantitative and time-dependent analysis of the exometabolome of Escherichia coli, Corynebacterium glutamicum, Bacillus licheniformis, and Saccharomyces cerevisiae during well-controlled bioreactor cultivations. Most surprisingly, in all cases a great diversity of central metabolic intermediates and amino acids is found in the culture medium with extracellular concentrations varying in the micromolar range. Different hypotheses for these observations are formulated and experimentally tested. As a result, the intermediates in the culture medium during batch growth must originate from passive or active transportation due to a new phenomenon termed "extended" overflow metabolism. Moreover, we provide broad evidence that this could be a common feature of all microorganism species when cultivated under conditions of carbon excess and non-inhibited carbon uptake. In turn, this finding has consequences for metabolite balancing and, particularly, for intracellular metabolite quantification and (13)C-metabolic flux analysis.
细胞overflow 代谢是酵母、细菌和哺乳动物细胞的一个众所周知的现象。它通常在葡萄糖过量的条件下发生,其特征是副产物如乙醇、乙酸或乳酸的排泄。这种现象也被称为短期 Crabtree 效应,在过去几十年中得到了广泛的研究,然而,其在代谢中的基本调节机制和功能作用仍然未知。在这里,我们对大肠杆菌、谷氨酸棒杆菌、地衣芽孢杆菌和酿酒酵母在严格控制的生物反应器培养过程中的外代谢组进行了全面的定量和时变分析。最令人惊讶的是,在所有情况下,在培养基中都发现了大量的中心代谢中间产物和氨基酸,其细胞外浓度在微摩尔范围内变化。对这些观察结果提出了不同的假设,并进行了实验验证。结果表明,由于一种新的现象称为“扩展”overflow 代谢,培养基中的中间产物必须来源于被动或主动运输。此外,我们提供了广泛的证据表明,当在碳过量和非抑制性碳摄取条件下培养时,这可能是所有微生物物种的共同特征。反过来,这一发现对代谢物平衡,特别是对细胞内代谢物定量和(13)C 代谢通量分析有影响。