Takahashi Hiroshi, Aihara Yuta, Ogawa Yukihiro, Murata Yoshimitsu, Nakajima Ken-Ichi, Iida Maiko, Shirai Miyako, Fujisaki Shingo
a Department of Biomolecular Science, Faculty of Science , Toho University , Chiba , Japan.
b Graduate School of Advanced Integration Science , Chiba University , Chiba , Japan.
Biosci Biotechnol Biochem. 2018 Jun;82(6):1003-1010. doi: 10.1080/09168451.2017.1398066. Epub 2017 Dec 1.
We investigated suppression of the slow growth of an Escherichia coli ispA null mutant lacking farnesyl diphosphate (FPP) synthase (i.e. IspA) by plasmids carrying prenyl diphosphate synthase genes. The growth rates of ispA mutant-transformants harboring a medium-copy number plasmid that carries ispA or ispB were almost the same as that of the wild-type strain. Although the level of FPP in the transformant with the ispA plasmid was almost the same as that in the wild-type strain, the level in the transformant with the ispB plasmid was as low as that in the ispA mutant. Purified octaprenyl diphosphate synthase (IspB) could condense isopentenyl diphosphate (IPP) with dimethylallyl diphosphate (DMAPP) to form octaprenyl diphosphate and nonaprenyl diphosphate. It is possible that suppression of the slow growth of the ispA mutant by ispB was due to condensation of IPP not only with FPP but also with DMAPP by octaprenyl diphosphate synthase.
我们研究了携带异戊二烯基二磷酸合酶基因的质粒对缺乏法尼基二磷酸(FPP)合酶(即IspA)的大肠杆菌ispA基因缺失突变体生长缓慢的抑制作用。携带ispA或ispB的中拷贝数质粒的ispA突变体转化子的生长速率与野生型菌株几乎相同。虽然携带ispA质粒的转化子中FPP的水平与野生型菌株中的几乎相同,但携带ispB质粒的转化子中FPP的水平与ispA突变体中的一样低。纯化的八聚异戊二烯基二磷酸合酶(IspB)可以将异戊烯基二磷酸(IPP)与二甲基烯丙基二磷酸(DMAPP)缩合形成八聚异戊二烯基二磷酸和九聚异戊二烯基二磷酸。ispB对ispA突变体生长缓慢的抑制作用可能是由于八聚异戊二烯基二磷酸合酶不仅使IPP与FPP缩合,还使其与DMAPP缩合。