Molecular biological analysis on protein secretion of koji mold

曲霉(Aspergillusoryzae)是一种重要的真菌，广泛应用于清酒、酱油、酱油等行业。它具有大量几种有用的酶的能力。本研究项目致力于有用蛋白的生产。结果1.克隆了在内质网分泌蛋白折叠中起重要作用的蛋白质二硫键异构酶基因，并对该基因的结构进行了分析。为了研究淀粉酶基因的诱导机制，比较了α-葡萄糖苷酶基因(AGDA)、α-淀粉酶基因(MyB)和糖酶基因(GLAA)，构建了启动子序列的缺失突变体。确定III区是淀粉酶诱导的重要区域，并开发了与III区串联的新型强启动子。克隆了与核分裂有关的肌动蛋白相关蛋白基因(ARPA)，并测定了其核苷酸序列。ARPA破碎物在菌核分裂中出现缺陷，菌丝超分枝形态异常。与WLID类型的菌株相比，这种形成许多菌丝顶端的干扰物分泌大量的α-淀粉酶。

Koji mold (Aspergillus oryzae) is one of the most important fungi, which has been used for sake, soy sauce, soy bean paste. It has an ability of a large amount of several useful enzymes. This resesarch project was fucused on the production of useful protein.1. A gene encoding protein disulfide isomerase, which Plays an important role in secreted protein foldind in ER,was cloned and the structure of the gene was analyzed.2. In order to examine the mechanism of induction of amylase genes, a comparison among alpha-glucosidase gene (agdA), alpha-amylase gene (amyB), and glucoamylase gene(glaA), construction of deletion mutants of the promoter sequence were done. Region III was identified as an important region for amylase induction and novel strong promoterwith tandem rpeats of the region III was developed.3. Actin related protein gene (arpA), which is involved in nuclear division, was cloned and determined its nucleotide sequence. The arpA disruptant showed defect in nucler division as well as an abnormal morphorogy of hyper-branched hyphae. This disruptant forming many hyphae tips secretes a large amount of alpha-amylase campared with its wlid type strain.