Umemura K, Atomi H, Kanai T, Teranishi Y, Ueda M, Tanaka A
Department of Synthetic Chemistry and Biological Chemistry, Faculty of Engineering, Kyoto University, Japan.
Appl Microbiol Biotechnol. 1995 Jul;43(3):489-92.
When the isocitrate lyase gene, containing 5'-upstream and 3'-flanking regions, of an n-alkane-assimilating yeast Candida tropicalis was introduced into Saccharomyces cerevisiae, the enzyme was functionally overexpressed in the cells grown on acetate. The amount of the recombinant isocitrate lyase expressed in S. cerevisiae was as much as 30% of the total soluble proteins in the cells, being comparable to that with GAL7 functional under the control of galactose. The expression was also observed when the cells were grown on glycerol, lactate, ethanol or oleate. These facts indicate that the isocitrate lyase gene upstream region (UPR-ICL) contains a strong promoter functional in S. cerevisiae. UPR-ICL is active as a promoter on cheap carbon sources such as acetate and nonconventional carbon sources such as oleate, whereas many conventional strong promoters demand relatively expensive sugars or sugar derivatives. Therefore, it is promising to construct an economical recombinant protein production system by using UPR-ICL.
将热带假丝酵母（一种正构烷烃同化酵母）的异柠檬酸裂解酶基因（包含5'-上游和3'-侧翼区域）导入酿酒酵母后，该酶在以乙酸盐为碳源生长的细胞中实现了功能性过表达。在酿酒酵母中表达的重组异柠檬酸裂解酶的量高达细胞总可溶性蛋白的30%，与在半乳糖控制下具有功能的GAL7相当。当细胞在甘油、乳酸、乙醇或油酸上生长时也观察到了这种表达。这些事实表明，异柠檬酸裂解酶基因上游区域（UPR-ICL）包含一个在酿酒酵母中具有功能的强启动子。UPR-ICL作为启动子在廉价碳源如乙酸盐和非常规碳源如油酸上具有活性，而许多传统的强启动子需要相对昂贵的糖类或糖衍生物。因此，利用UPR-ICL构建经济的重组蛋白生产系统具有广阔前景。
