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在含有多个拷贝agdA基因的米曲霉转化体中,三个淀粉酶编码基因agdA、amyB和glaA的特征性表达。

Characteristic expression of three amylase-encoding genes, agdA, amyB, and glaA in Aspergillus oryzae transformants containing multiple copies of the agdA gene.

作者信息

Minetoki T, Gomi K, Kitamoto K, Kumagai C, Tamura G

机构信息

Research Institute of Brewing Resources Co., Ltd., Tokyo, Japan.

出版信息

Biosci Biotechnol Biochem. 1995 Dec;59(12):2251-4. doi: 10.1271/bbb.59.2251.

Abstract

In Aspergillus oryzae wild-type strains, the expression of the agdA gene encoding alpha-glucosidase (AGL) is induced by maltose at the transcriptional level in a similar manner to the amyB gene encoding Takaamylase A (TAA) and the glaA gene encoding glucoamylase (GLA). In A. oryzae transformants containing multiple copies of the agdA gene, a high-level of AGL activity was observed. This was accompanied by a significant reduction in TAA and GLA activities. Moreover, transformants with the highest AGL activity showed the lowest degree of TAA and GLA activities. Northern blot analyses showed that the transcriptional levels of amyB and glaA in the AGL-overproducing transformant were drastically reduced when large amounts of agdA mRNA were detected in maltose-grown mycelia. In addition, the glucose concentration of the maltose-containing medium that was used to grow the AGL-overproducing transformant for RNA extraction was higher than that of the control transformant. These results suggest that the reduced expression of the amyB and glaA genes in the AGL-overproducing transformant was due to either titration of a common regulatory protein(s) involved in maltose induction or carbon catabolite repression.

摘要

在米曲霉野生型菌株中,编码α-葡萄糖苷酶(AGL)的agdA基因的表达在转录水平上由麦芽糖诱导,其方式与编码米曲霉淀粉酶A(TAA)的amyB基因和编码糖化酶(GLA)的glaA基因相似。在含有多个agdA基因拷贝的米曲霉转化体中,观察到高水平的AGL活性。这伴随着TAA和GLA活性的显著降低。此外,AGL活性最高的转化体显示出最低程度的TAA和GLA活性。Northern印迹分析表明,当在麦芽糖培养的菌丝体中检测到大量agdA mRNA时,AGL过量产生的转化体中amyB和glaA的转录水平急剧降低。此外,用于培养AGL过量产生的转化体以提取RNA的含麦芽糖培养基的葡萄糖浓度高于对照转化体。这些结果表明,AGL过量产生的转化体中amyB和glaA基因表达的降低是由于参与麦芽糖诱导的共同调节蛋白的滴定或碳分解代谢物阻遏。

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