Robbins J C, Oxender D L
J Bacteriol. 1973 Oct;116(1):12-8. doi: 10.1128/jb.116.1.12-18.1973.
At least two transport systems serve for the entry of alanine, glycine, and serine into Escherichia coli. One of these systems serves mainly for glycine, d-alanine, and d-serine and to some extent for l-alanine, whereas the second serves for l-alanine and perhaps l-serine. These two transport systems have been characterized by kinetic studies and by inhibition analysis. Reciprocal plots for l-alanine entry are distinctly biphasic, giving rise to K(m) values of about 2 and 27 muM. The major route of glycine entry can be described by a single K(m) value of about 4 muM. A higher K(m) value for glycine of around 70 to 100 muM shows that other routes of entry may serve at high concentrations of amino acid. The glycine, d-serine and d-alanine transport system is defective in a d-serine-resistant mutant, strain EM1302. The mutation, dagA, is recessive in dagA/dagA(+) merodiploids and is 7 to 12% linked by phage P1 transduction to the pyrB locus of E. coli. E. coli with the dagA mutation are unable to utilize d-alanine as a carbon source, providing an additional basis for selecting such mutants. The remaining l-alanine uptake in dagA mutants is subject to inhibition by l-serine, l-threonine, and l-leucine. It is also sensitive to osmotic shock treatment and repressed by growth of the cells on l-leucine. It appears from a comparison of the properties of the second l-alanine system with those of the leucine, isoleucine, and valine system (LIV system) that the LIV system also serves for the transport of l-alanine and l-threonine and perhaps l-serine.
至少有两种转运系统负责将丙氨酸、甘氨酸和丝氨酸转运到大肠杆菌中。其中一种系统主要负责转运甘氨酸、d-丙氨酸和d-丝氨酸,在一定程度上也负责转运l-丙氨酸;而另一种系统则负责转运l-丙氨酸,可能还包括l-丝氨酸。这两种转运系统已通过动力学研究和抑制分析进行了表征。l-丙氨酸进入细胞的双倒数作图明显呈双相性,产生的K(m)值约为2和27 μM。甘氨酸进入细胞的主要途径可用约4 μM的单一K(m)值来描述。甘氨酸在70至100 μM左右的较高K(m)值表明,在高浓度氨基酸情况下可能存在其他进入途径。甘氨酸、d-丝氨酸和d-丙氨酸转运系统在d-丝氨酸抗性突变体EM1302菌株中存在缺陷。该突变基因dagA在dagA/dagA(+)部分二倍体中是隐性的,通过噬菌体P1转导与大肠杆菌的pyrB基因座有7%至12%的连锁。带有dagA突变的大肠杆菌无法利用d-丙氨酸作为碳源,这为筛选此类突变体提供了额外的依据。dagA突变体中剩余的l-丙氨酸摄取受到l-丝氨酸、l-苏氨酸和l-亮氨酸的抑制。通过比较第二种l-丙氨酸系统与亮氨酸、异亮氨酸和缬氨酸系统(LIV系统)的特性发现,LIV系统也负责转运l-丙氨酸和l-苏氨酸,可能还包括l-丝氨酸。