Estévez A M, Heinisch J J, Aragón J J
Departamento de Bioquímica de la UAM, Facultad de Medicina, Universidad Autónoma, Madrid, Spain.
FEBS Lett. 1995 Oct 23;374(1):100-4. doi: 10.1016/0014-5793(95)01085-s.
Phosphofructokinase (PFK) from yeast has been replaced by the non-allosteric isozyme from the slime mold Dictyostelium discoideum. This has been achieved by overexpression of the latter in a PFK-deficient strain of Saccharomyces cerevisiae under the control of the PFK2 promoter. Transformants complemented the glucose-negative growth phenotype exhibiting generation times on glucose-containing media similar to those of an untransformed strain being wild-type for yeast PFK genes. The PFK produced reacted with an antibody against D. discoideum PFK. It exhibited the same subunit size, quaternary structure and kinetic parameters than those of the wild-type enzyme, and was also devoid of specific regulatory properties.
酵母中的磷酸果糖激酶（PFK）已被黏菌盘基网柄菌的非别构同工酶所取代。这是通过在PFK2启动子的控制下，在酿酒酵母的PFK缺陷菌株中过表达后者来实现的。转化体弥补了葡萄糖阴性生长表型，在含葡萄糖的培养基上的代时与未转化的酵母PFK基因野生型菌株相似。产生的PFK与抗盘基网柄菌PFK的抗体发生反应。它表现出与野生型酶相同的亚基大小、四级结构和动力学参数，并且也没有特定的调节特性。
