Department of Food Science and Biotechnology, Faculty of Chemistry, Universidad Nacional Autónoma de México (UNAM), Ciudad Universitaria, México City, DF 04510, Mexico.
Appl Biochem Biotechnol. 2013 Mar;169(5):1672-82. doi: 10.1007/s12010-013-0091-8. Epub 2013 Jan 19.
Aspergillus nidulans produces several proteases. The prtA gene encodes a major protease, and two approaches were explored to achieve the overproduction of this enzyme. Molecular cloning of the mature form of this enzyme in Pichia pastoris resulted in the production of an inactive form. In addition, the presence of this enzyme was toxic for the host and resulted in cell lysis. The modification of the culture medium constituents resulted in a 6.4-fold increase in enzyme production. The main effect was achieved through the use of organic nitrogen sources. Although it was previously shown that the PrtA protease shows promiscuous esterase activity, the production of this enzyme was not induced by lipidic sources.
构巢曲霉能产生多种蛋白酶。prtA 基因编码一种主要的蛋白酶，我们探索了两种方法来实现该酶的过量生产。在巴斯德毕赤酵母中对该酶成熟形式进行分子克隆导致产生了无活性的形式。此外，该酶的存在对宿主有毒，导致细胞裂解。通过对培养基成分的修饰，使酶的产量增加了 6.4 倍。主要效果是通过使用有机氮源实现的。尽管之前已经表明 PrtA 蛋白酶具有混杂的酯酶活性，但这种酶的产生并没有被脂质源诱导。
