Weisser P, Krämer R, Sahm H, Sprenger G A
Institut für Biotechnologie, Forschungszentrum Jülich GmbH, Germany.
J Bacteriol. 1995 Jun;177(11):3351-4. doi: 10.1128/jb.177.11.3351-3354.1995.
The Zymomonas mobilis genes encoding the glucose facilitator (glf), glucokinase (glk), or fructokinase (frk) were cloned and expressed in a lacIq-Ptac system using Escherichia coli K-12 mutants deficient in uptake and phosphorylation of glucose and fructose. Growth on glucose or fructose was restored when the respective genes (glf-glk or glf-frk) were expressed. In E. coli glf+ strains, both glucose and fructose were taken up via facilitated diffusion (Km, 4.1 mM for glucose and 39 mM for fructose; Vmax at 15 degrees C, 75 and 93 nmol min-1 mg-1 [dry weight] for glucose and fructose, respectively). For both substrates, counterflow maxima were observed.
编码葡萄糖转运蛋白(glf)、葡萄糖激酶(glk)或果糖激酶(frk)的运动发酵单胞菌基因,利用缺乏葡萄糖和果糖摄取及磷酸化能力的大肠杆菌K - 12突变体,在lacIq - Ptac系统中进行克隆和表达。当相应基因(glf - glk或glf - frk)表达时,在葡萄糖或果糖上的生长得以恢复。在大肠杆菌glf⁺菌株中,葡萄糖和果糖均通过易化扩散摄取(葡萄糖的Km为4.1 mM,果糖为39 mM;15℃时葡萄糖和果糖的Vmax分别为75和93 nmol min⁻¹ mg⁻¹[干重])。对于这两种底物,均观察到逆向流动最大值。