Efficient expression and secretion of Aspergillus niger RH5344 polygalacturonase in Saccharomyces cerevisiae
Efficient expression and secretion of Aspergillus niger RH5344 polygalacturonase in Saccharomyces cerevisiae
复制标题
黑曲霉RH5344多聚半乳糖醛酸酶在酿酒酵母中的高效表达和分泌
DOI：
10.1007/bf00164494
复制
发表时间：
1995
影响因子：
5
通讯作者：
A. Looman
中科院分区：
文献类型：
--
作者：
C. Lang;A. Looman
尼日尔曲霉内切多聚半乳糖醛酸酶（EC 3.2.1.15）cDNA在酵母酿酒酵母中表达。蛋白质分泌到生长培养基中是由真菌前导序列有效地指导的，并且加工发生在与曲霉菌相同的位点。通过使用酵母ADHI启动子的“短”版本，表达水平显著增强。异源蛋白产量的额外增加是由于质粒稳定性更高和质粒拷贝数增加。这是通过从表达载体中删除大部分细菌序列来实现的。该酶具有与真菌多聚半乳糖醛酸酶相同的酶学和生化特性，如底物特异性、最适pH和温度以及pI值。然而，酵母衍生的酶显示出更高的糖基化程度，并表现出比真菌酶更显著的温度稳定性。
An Aspergillus niger endopolygalacturonase (EC 3.2.1.15) cDNA was expressed in the yeast Saccharomyces cerevisiae. Secretion of the protein into the growth medium was efficiently directed by the fungal leader sequence, and processing occurred at the same site as in Aspergillus. The expression level was significantly enhanced by using a “short” version of the yeast ADHI promoter. An additional increase in the yield of heterologous protein was due to a higher plasmid stability and a rise in plasmid copy number. This was achieved by deleting most of the bacterial sequences from the expression vector. The yeast-derived enzyme showed the same enzymatic and biochemical properties as the fungal polygalacturonase, such as substrate specificity, pH and temperature optima and pI value. The yeast-derived enzyme, however, showed a higher degree of glycosylation and exhibited a more pronounced temperature stability than the fungal enzyme.
DOI：
10.1016/s0021-9258(18)33174-0
发表时间：
1983-01
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
C. Denis;J. Ferguson;E. Young
通讯作者：
C. Denis;J. Ferguson;E. Young
DOI：
10.1016/s0021-9258(19)39664-4
发表时间：
1990-03
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
K. Frederick;J. Tung;R. Emerick;F. R. Masiarz;S H Chamberlain;A. Vasavada;S. Rosenberg;S. Chakraborty;L. Schopfer;L. M. Schopter
通讯作者：
K. Frederick;J. Tung;R. Emerick;F. R. Masiarz;S H Chamberlain;A. Vasavada;S. Rosenberg;S. Chakraborty;L. Schopfer;L. M. Schopter