Mols J, Peeters-Joris C, Agathos S N, Schneider Y-J
Laboratoire de Biochimie cellulaire, Place Louis Pasteur, 1., Belgium.
Biotechnol Lett. 2004 Jul;26(13):1043-6. doi: 10.1023/B:BILE.0000032960.06112.31.
CHO-320 cells, cultivated in suspension in a protein-free medium supplemented with rice protein hydrolysates (peptones), secrete recombinant interferon-gamma (IFN-gamma) that undergo will or will not proteolysis, depending on the origin of the peptones. This proteolytic event, as well as the appearance of an unidentified 70 kDa gelatinase-like protease, are attributed to a cysteine protease. Casein zymographies revealed that one rice protein hydrolysate, but not another, contains a papain-like cysteine protease whose activity is undetectable in solution. This work underlines the significance of the origin of peptones when considered as supplements in serum- and protein-free media for overproduction of recombinant proteins.
CHO-320细胞在添加了大米蛋白水解物(蛋白胨)的无蛋白培养基中悬浮培养,分泌重组干扰素-γ(IFN-γ),根据蛋白胨的来源,该重组干扰素-γ会发生或不会发生蛋白水解。这种蛋白水解事件以及一种未鉴定的70 kDa明胶酶样蛋白酶的出现,都归因于一种半胱氨酸蛋白酶。酪蛋白酶谱分析表明,一种大米蛋白水解物含有一种木瓜蛋白酶样半胱氨酸蛋白酶,而另一种则不含,其活性在溶液中无法检测到。这项工作强调了在用于重组蛋白过量生产的无血清和无蛋白培养基中,将蛋白胨作为补充剂时其来源的重要性。