Harold F M, Levin E
J Bacteriol. 1974 Mar;117(3):1141-8. doi: 10.1128/jb.117.3.1141-1148.1974.
Streptococcus faecalis obtains metabolic energy chiefly from the conversion of glucose to lactic acid; the present experiments deal with the mechanism of lactic acid translocation across the cytoplasmic membrane. Efflux of [(14)C]lactate from preloaded cells was accelerated by raising the external pH, and also by the ionophores nigericin and valinomycin. These results suggest that lactate leaves the cell by an electroneutral process, presumably as lactic acid. Further evidence was obtained by studying the entry of [(14)C]lactate into nonmetabolizing cells. It appears that the membrane is essentially impermeable to the lactate anion, but allows passage of lactic acid. The most persuasive evidence is that, upon establishment of a pH gradient such that the cytoplasm was alkaline, l-[(14)C]lactate accumulated in the cells against the concentration gradient. Accumulation was transient, and dissipated in parallel with the collapse of the pH gradient. The concentration gradient attained at the peak was a function of the pH difference. Ionophores which are known to collapse a pH gradient, such as nigericin and valinomycin, abolished accumulation of l-lactate. We infer that lactic acid translocation, whether into the cells or outward, is an electroneutral process and for that reason the distribution of lactic acid across the membrane is a function of the pH of cytoplasm and medium. The specificity of translocation and its kinetic parameters suggest that it is mediated by a carrier of low specificity.
粪肠球菌主要通过将葡萄糖转化为乳酸来获取代谢能量;目前的实验研究了乳酸跨细胞质膜转运的机制。预先加载细胞中的[¹⁴C]乳酸外流通过提高细胞外pH值以及离子载体尼日利亚菌素和缬氨霉素而加速。这些结果表明,乳酸通过电中性过程离开细胞,推测是以乳酸的形式。通过研究[¹⁴C]乳酸进入非代谢细胞的过程获得了进一步的证据。看来细胞膜对乳酸阴离子基本不可渗透,但允许乳酸通过。最有说服力的证据是,当建立起细胞质呈碱性的pH梯度时,L-[¹⁴C]乳酸在细胞内逆浓度梯度积累。积累是短暂的,并随着pH梯度的消失而消散。峰值时达到的浓度梯度是pH差值的函数。已知能消除pH梯度的离子载体,如尼日利亚菌素和缬氨霉素,会消除L-乳酸的积累。我们推断,乳酸的转运,无论是进入细胞还是向外转运,都是一个电中性过程,因此乳酸在膜两侧的分布是细胞质和培养基pH值的函数。转运的特异性及其动力学参数表明它是由低特异性的载体介导的。