Goel Anisha, Eckhardt Thomas H, Puri Pranav, de Jong Anne, Branco Dos Santos Filipe, Giera Martin, Fusetti Fabrizia, de Vos Willem M, Kok Jan, Poolman Bert, Molenaar Douwe, Kuipers Oscar P, Teusink Bas
Systems Bioinformatics IBIVU, Vrije Universiteit Amsterdam, De Boelelaan 1085, 1081 HV, Amsterdam, The Netherlands.
Laboratory of Microbiology, Wageningen University, Dreijenplein 10, 6703 HB, Wageningen, The Netherlands.
Mol Microbiol. 2015 Jul;97(1):77-92. doi: 10.1111/mmi.13012. Epub 2015 May 9.
Protein investment costs are considered a major driver for the choice of alternative metabolic strategies. We tested this premise in Lactococcus lactis, a bacterium that exhibits a distinct, anaerobic version of the bacterial Crabtree/Warburg effect; with increasing growth rates it shifts from a high yield metabolic mode [mixed-acid fermentation; 3 adenosine triphosphate (ATP) per glucose] to a low yield metabolic mode (homolactic fermentation; 2 ATP per glucose). We studied growth rate-dependent relative transcription and protein ratios, enzyme activities, and fluxes of L. lactis in glucose-limited chemostats, providing a high-quality and comprehensive data set. A three- to fourfold higher growth rate rerouted metabolism from acetate to lactate as the main fermentation product. However, we observed hardly any changes in transcription, protein levels and enzyme activities. Even levels of ribosomal proteins, constituting a major investment in cellular machinery, changed only slightly. Thus, contrary to the original hypothesis, central metabolism in this organism appears to be hardly regulated at the level of gene expression, but rather at the metabolic level. We conclude that L. lactis is either poorly adapted to growth at low and constant glucose concentrations, or that protein costs play a less important role in fitness than hitherto assumed.
蛋白质投资成本被认为是选择替代代谢策略的主要驱动因素。我们在乳酸乳球菌中验证了这一前提,该细菌表现出独特的、厌氧形式的细菌Crabtree/瓦伯格效应;随着生长速率的增加,它从高产代谢模式[混合酸发酵;每葡萄糖产生3个三磷酸腺苷(ATP)]转变为低产代谢模式(同型乳酸发酵;每葡萄糖产生2个ATP)。我们研究了葡萄糖限制恒化器中乳酸乳球菌的生长速率依赖性相对转录和蛋白质比率、酶活性及通量,提供了一个高质量且全面的数据集。高三到四倍的生长速率将代谢从乙酸盐重新导向乳酸盐,使其成为主要发酵产物。然而,我们几乎未观察到转录、蛋白质水平和酶活性有任何变化。甚至构成细胞机制主要投资的核糖体蛋白水平也仅有轻微变化。因此,与最初的假设相反,该生物体的中心代谢似乎在基因表达水平上几乎不受调控,而是在代谢水平上受到调控。我们得出结论,乳酸乳球菌要么对低且恒定的葡萄糖浓度下的生长适应性较差,要么蛋白质成本在适应性方面所起的作用比迄今所认为的要小。