Molecular biological analysis on protein secretion of koji mold

曲霉菌蛋白质分泌的分子生物学分析

• 批准号: 08456045
• 负责人: KITAMOTO Katsuhiko
• 金额: $ 4.54万
• 依托单位: The University of Tokyo
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1996
• 资助国家: 日本
• 起止时间: 1996 至 1997
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08456045/
• 关键词: Koji mold; protein secretion; molecular biology; protein disulfide isomerase; alpha-glucosidase; Aspergillus

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.

曲霉（Aspergillus oryzae）是最重要的真菌之一，已被用于清酒、酱油、豆沙。它具有大量几种有用酶的能力。这个研究项目的重点是有用蛋白的生产。克隆了一个在内质网分泌蛋白折叠中起重要作用的编码蛋白二硫异构酶的基因，并分析了该基因的结构。为了探讨淀粉酶基因的诱导机制，对α -葡萄糖苷酶基因（agdA）、α -淀粉酶基因（amyB）和葡萄糖淀粉酶基因（glaA）进行了比较，构建了启动子序列缺失突变体。III区被认为是淀粉酶诱导的重要区域，并开发了具有串联重复序列的新型强启动子。克隆了参与核分裂的肌动蛋白相关蛋白基因（Actin related protein gene, arpA），并测定了其核苷酸序列。arpA干扰物表现为细胞核分裂缺陷和菌丝超支化形态异常。与野生型菌株相比，这种形成许多菌丝尖端的破坏物分泌大量的α -淀粉酶。

项目成果

T.Minetoki: "Deletion Anaoysis of promoter Elements of the Aspergillus oryzae agdA Gene Encoding α-glucosidase" Curr.Genet. 30. 432-438 (1996)

T.Minetoki：“米曲霉 agdA 基因编码 α-葡萄糖苷酶的启动子元件的删除分析”Curr.Genet. 30. 432-438 (1996)

T M,netok,: "Deletion Analysis of Promoter Elements of the Aspergillus eryzae agdA gene Encoding α-glucosidase" Curr Genet.30. 432-438 (1996)

T M，netok，：“编码 α-葡萄糖苷酶的米曲霉 agdA 基因的启动子元件的删除分析”Curr Genet.30 (1996)。

北本勝ひこ: "麹菌(A,oryzae)の分子生物学的解析の最近の進展" 日本農芸化学会誌. 71. 27-28 (1997)

Katsuhiko Kitamoto：“米曲霉分子生物学分析的最新进展”日本农业化学学会杂志 71. 27-28 (1997)。

T.Minetoki, and K.Kitamoto: "Gene analysis and role of rice koji in sake brewing" Nippon Nogeikagaku kaishi. 71. 34-39 (1998)

T.Minetoki 和 K.Kitamoto：“基因分析和米曲在清酒酿造中的作用”Nippon Nogeikagaku kaishi。

B.R.Lee: "Cloning Characterization and Overexpression of a Gene(pdi A) Encoding Protein Disulfide Isomerase of Aspergillus oryzae" J.Ferment.Bioeng.82. 538-543 (1996)

B.R.Lee：“编码米曲霉蛋白二硫键异构酶的基因 (pdi A) 的克隆特征和过表达”J.Ferment.Bioeng.82。