Szkopińska A, Rytka J, Karst F, Palamarczyk G
Institute of Biochemistry and Biophysics PAN, Warsaw, Poland.
FEMS Microbiol Lett. 1993 Sep 15;112(3):325-8. doi: 10.1111/j.1574-6968.1993.tb06470.x.
Mutants deficient in sterol (thermosensitive ergosterol auxotrophs) erg 8, 9, 12 and heme synthesis hem 1, 12 were screened for the level of free dolichol and dolichyl phosphate synthesized in the mevalonate pathway as well as for the activity of dolichyl phosphate-dependent glycosyl transferases. The amount of DolP synthesized via CTP-dependent phosphorylation was the same in mutants and parental strains. However, mannosylation and glucosylation of endogenous dolichyl phosphates in ergosterol mutants was about four times lower compared to parental strains, while the same reactions carried out with exogenous Dol24P reached 80% of the level observed in parental strains indicating that activities of DolPMan and DolPGlc synthases are not the rate-limiting factors. It is postulated that the de novo synthesis of DolP is impaired in the ergosterol mutants. Moreover, a block in the ergosterol branch of the metabolic pathway (erg 9) causes an increase in the de novo synthesis of dolichyl phosphate.
对甾醇缺陷型(温度敏感型麦角甾醇营养缺陷型)erg 8、9、12以及血红素合成缺陷型hem 1、12进行筛选,检测其在甲羟戊酸途径中合成的游离多萜醇和多萜醇磷酸酯的水平,以及多萜醇磷酸酯依赖性糖基转移酶的活性。通过CTP依赖性磷酸化合成的DolP量在突变体和亲本菌株中相同。然而,与亲本菌株相比,麦角甾醇突变体内源性多萜醇磷酸酯的甘露糖基化和葡萄糖基化降低了约四倍,而用外源性Dol24P进行的相同反应达到了亲本菌株中观察到水平的80%,这表明DolPMan和DolPGlc合酶的活性不是限速因素。据推测,麦角甾醇突变体中DolP的从头合成受到损害。此外,代谢途径中麦角甾醇分支(erg 9)的阻断导致多萜醇磷酸酯的从头合成增加。