High-level secretion of a Rhizopus niveus aspartic proteinase in Saccharomyces cerevisiae.
High-level secretion of a Rhizopus niveus aspartic proteinase in Saccharomyces cerevisiae.
复制标题
酿酒酵母中雪根霉天冬氨酸蛋白酶的高水平分泌。
DOI：
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发表时间：
1990
期刊：
影响因子：
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通讯作者：
K. Yano
中科院分区：
文献类型：
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作者：
H. Horiuchi;T. Ashikari;T. Amachi;H. Yoshizumi;M. Takagi;K. Yano
将编码细胞外雪根霉天冬氨酸蛋白酶 I (RNAP-I) 的基因引入酿酒酵母中。携带含有在酿酒酵母甘油醛3-磷酸脱氢酶(GAPDH)基因启动子控制下的完整RNAP-I基因的质粒的酵母细胞根本不合成RNAP-I。另一方面，当RNAP-I基因的内含子已从质粒中的基因中去除时，细胞高效地分泌RNAP-I。在培养过程中，前序列的加工发生在与在雪白红葡萄球菌培养物中观察到的相同的酶原区域。此外，发现原始RNAP-I基因的启动子和终止子在酵母细胞中对于无内含子RNAP-I基因的表达而言功能较弱，尽管起始位点和终止位点是异质的。还研究了载体类型对 RNAP-I 细胞外产生的影响。
The gene encoding an extracellular Rhizopus niveus aspartic proteinase I (RNAP-I) was introduced into Saccharomyces cerevisiae. The yeast cell carrying a plasmid containing the intact RNAP-I gene under the control of the glyceraldehyde 3-phosphate dehydrogenase (GAPDH) gene promoter of S. cerevisiae did not synthesize RNAP-I at all. On the other hand, when the intron of the RNAP-I gene had been removed from the gene in the plasmid, the cell secreted RNAP-I with high efficiency. Processing of the pro-sequence occurred at the same region of the pro-enzyme during cultivation as observed in the culture of R. niveus. Moreover, the promoter and the terminator of the original RNAP-I gene were found to be weakly functional in the yeast cell with respect to expression of the intronless RNAP-I gene, although the initiation and termination sites were heterogeneous. The effects of vector-types on the extracellular production of RNAP-I were also investigated.