Paifer E, Margolles E, Cremata J, Montesino R, Herrera L, Delgado J M
Center of Genetic Engineering & Biotechnology, Division of Industrial Biotechnology, Habana, Cuba.
Yeast. 1994 Nov;10(11):1415-9. doi: 10.1002/yea.320101104.
We have cloned and expressed a bacterial thermostable alpha amylase gene in Pichia pastoris using the methanol-controlled alcohol oxidase (AOX1) promoter. Two integrative vectors were constructed with two different secretion signal sequences in order to obtain efficient secretion of the protein. One vector contains the structural gene encoding the mature alpha amylase fused to the SUC2 gene signal sequence from Saccharomyces cerevisiae. In the other vector, the alpha amylase is expressed with its own signal sequence. In both cases, the alpha amylase were secreted into the culture medium with high efficiency, around 2.5 and 0.9 g/l respectively.
我们利用甲醇控制的醇氧化酶（AOX1）启动子，在巴斯德毕赤酵母中克隆并表达了一个细菌耐热α淀粉酶基因。构建了两个带有不同分泌信号序列的整合载体，以便高效分泌该蛋白。一个载体包含编码成熟α淀粉酶的结构基因，该基因与来自酿酒酵母的SUC2基因信号序列融合。在另一个载体中，α淀粉酶以其自身的信号序列进行表达。在这两种情况下，α淀粉酶均高效分泌到培养基中，分别约为2.5 g/l和0.9 g/l。
