Koebmann Brian, Blank Lars Mathias, Solem Christian, Petranovic Dina, Nielsen Lars K, Jensen Peter Ruhdal
Fermenco ApS, Technical University of Denmark, Building 301, DK-2800 Kgs. Lyngby, Denmark.
Biotechnol Appl Biochem. 2008 May;50(Pt 1):25-33. doi: 10.1042/BA20070132.
Lactococcus lactis is known to be capable of respiration under aerobic conditions in the presence of haemin. In the present study the effect of respiration on ATP production during growth on different sugars was examined. With glucose as the sole carbon source, respiratory conditions in L. lactis MG1363 resulted in only a minor increase, 21%, in biomass yield. Since ATP production through substrate-level phosphorylation was essentially identical with and without respiration, the increased biomass yield was a result of energy-saving under respiratory conditions estimated to be 0.4 mol of ATP/mol of glucose. With maltose as the energy source, the increase in biomass yield amounted to 51% compared with an aerobic culture that lacked haemin. This higher ATP yield was obtained by redirecting pyruvate metabolism from lactate to acetate production, and from savings through respiration. However, even after subtracting these contributions, approx. 0.3 mol of ATP/mol of glucose remained unaccounted for. A similar response to respiratory conditions (0.2 mol of ATP/mol of glucose) was observed in a mutant that had a decreased glucose uptake rate during growth on glucose caused by disruption of the PTS(mannose) (glucose/mannose-specific phosphotransferase system). Amino acid catabolism could be excluded as the source of the additional ATP. Since mutants without a functional H+-ATPase produced less ATP under sugar starvation and respiratory conditions, the additional ATP yield appears to come partly from energy saved on proton pumping through the H+-ATPase due to respiration and partly from a reversed function of the H+-ATPase towards oxidative phosphorylation. These results may contribute to the design and implementation of carbon-efficient high-cell-density cultures of this industrially important species of bacterium.
已知乳酸乳球菌在有氧条件下,存在血红素时能够进行呼吸作用。在本研究中,检测了呼吸作用对不同糖类生长过程中ATP产生的影响。以葡萄糖作为唯一碳源时,乳酸乳球菌MG1363中的呼吸条件仅使生物量产量略有增加,即增加了21%。由于通过底物水平磷酸化产生ATP在有呼吸和无呼吸条件下基本相同,生物量产量的增加是由于在呼吸条件下节省了能量,估计为每摩尔葡萄糖节省0.4摩尔ATP。以麦芽糖作为能源时,与缺乏血红素的需氧培养相比,生物量产量增加了51%。通过将丙酮酸代谢从乳酸生产转向乙酸生产以及通过呼吸节省能量,获得了更高的ATP产量。然而,即使减去这些贡献,仍有约每摩尔葡萄糖0.3摩尔ATP无法解释。在一个突变体中观察到对呼吸条件的类似反应(每摩尔葡萄糖0.2摩尔ATP),该突变体由于PTS(甘露糖)(葡萄糖/甘露糖特异性磷酸转移酶系统)的破坏,在以葡萄糖生长期间葡萄糖摄取率降低。氨基酸分解代谢可被排除为额外ATP的来源。由于没有功能性H + -ATP酶的突变体在糖饥饿和呼吸条件下产生的ATP较少,额外的ATP产量似乎部分来自于由于呼吸作用通过H + -ATP酶进行质子泵浦时节省的能量,部分来自于H + -ATP酶对氧化磷酸化的反向作用。这些结果可能有助于设计和实施这种具有重要工业价值的细菌的碳高效高细胞密度培养。