Im D S, Friedrich C G
J Bacteriol. 1983 May;154(2):803-8. doi: 10.1128/jb.154.2.803-808.1983.
Alcaligenes eutrophus formed ribulosebisphosphate carboxylase (RuBPCase; EC 4.1.1.39) when grown on fructose. Addition of sodium fluoride (NaF) to fructose minimal medium resulted in a slightly decreased growth rate and a rapid fivefold increase in RuBPCase specific activity. With citrate, a glucogenic carbon source, RuBPCase was also formed, However, addition of NaF to cells growing on citrate resulted in a 50% decrease in RuBPCase specific activity. Among the enzymes of fructose catabolism, NaF (10 mM) inhibited enolase in vitro by 98% and gluconate 6-phosphate dehydratase by 87%. Inhibition of the dehydratase by NaF was insignificant in vivo, as determined with a mutant defective in phosphoglycerate mutase activity. Growth of this mutant on fructose was not inhibited by NaF, and only a minor increase in RuBPCase activity was observed. From these results, we concluded that the product of the enolase reaction, phosphoenolpyruvate, played a role in RuBPCase formation. Addition of H2 or formate to the wild type growing on fructose or citrate did not affect the growth rate but resulted in rapid formation of RuBPCase activity. Mutants impaired in H2 metabolism formed RuBPCase at a low rate during growth on fructose plus H2 but at a high rate on formate. Apparently, additional reductant from H2 or formate metabolism induced RuBPCase formation in A. eutrophus.
嗜糖产碱菌在以果糖为碳源生长时会形成核酮糖-1,5-二磷酸羧化酶(RuBPCase;EC 4.1.1.39)。向果糖基本培养基中添加氟化钠(NaF)会导致生长速率略有下降,但RuBPCase的比活性会迅速增加五倍。以生糖碳源柠檬酸盐为底物时,也会形成RuBPCase。然而,向以柠檬酸盐为碳源生长的细胞中添加NaF会导致RuBPCase的比活性降低50%。在果糖分解代谢的酶中,NaF(10 mM)在体外对烯醇化酶的抑制率为98%,对6-磷酸葡萄糖酸脱水酶的抑制率为87%。用磷酸甘油酸变位酶活性有缺陷的突变体进行测定发现,NaF在体内对脱水酶的抑制作用不明显。该突变体在果糖上的生长不受NaF抑制,仅观察到RuBPCase活性略有增加。根据这些结果,我们得出结论,烯醇化酶反应的产物磷酸烯醇丙酮酸在RuBPCase的形成中起作用。向以果糖或柠檬酸盐为碳源生长的野生型菌株中添加H2或甲酸盐,不会影响生长速率,但会导致RuBPCase活性迅速形成。H2代谢受损的突变体在果糖加H2的条件下生长时,RuBPCase的形成速率较低,但在甲酸盐条件下形成速率较高。显然,来自H2或甲酸盐代谢的额外还原剂会诱导嗜糖产碱菌中RuBPCase的形成。