Schaaff I, Hohmann S, Zimmermann F K
Institut für Mikrobiologie, Technische Hochschule Darmstadt, Federal Republic of Germany.
Eur J Biochem. 1990 Mar 30;188(3):597-603. doi: 10.1111/j.1432-1033.1990.tb15440.x.
We have cloned the structural gene for yeast transaldolase. Transformants carrying the TAL1 gene on a multicopy plasmid over-produced transaldolase. A deletion mutant which was constructed using the cloned gene did not show any detectable transaldolase activity in vitro. Furthermore, both transaldolase isoenzymes which were detected in wild-type crude extracts by immunoblotting were missing in the deletion mutants. Thus, TAL1 is the only transaldolase structural gene in yeast. TAL1 is not an essential gene. Deletion of the transaldolase gene did not affect growth on complete media with different carbon sources or on synthetic media. However, the transaldolase-deficient strains accumulated sedoheptulose 7-phosphate, an intermediate of the pentose-phosphate pathway. Mutants lacking both transaldolase and phosphoglucose isomerase grew more slowly than the single mutants. They accumulated more sedoheptulose 7-phosphate on medium containing fructose than on glucose medium. This shows that fructose 6-phosphate and glyceraldehyde 3-phosphate, metabolites of glycolysis, can enter the nonoxidative part of the pentose-phosphate pathway.
我们已经克隆了酵母转醛醇酶的结构基因。携带多拷贝质粒上TAL1基因的转化子过量产生转醛醇酶。利用克隆基因构建的缺失突变体在体外未显示出任何可检测到的转醛醇酶活性。此外,通过免疫印迹在野生型粗提物中检测到的两种转醛醇酶同工酶在缺失突变体中均不存在。因此,TAL1是酵母中唯一的转醛醇酶结构基因。TAL1不是必需基因。转醛醇酶基因的缺失并不影响在含有不同碳源的完全培养基或合成培养基上的生长。然而,转醛醇酶缺陷型菌株积累了景天庚酮糖7-磷酸,这是戊糖磷酸途径的一种中间产物。缺乏转醛醇酶和磷酸葡萄糖异构酶的突变体比单突变体生长得更慢。它们在含有果糖的培养基上比在葡萄糖培养基上积累更多的景天庚酮糖7-磷酸。这表明糖酵解的代谢产物6-磷酸果糖和3-磷酸甘油醛可以进入戊糖磷酸途径的非氧化部分。