Samuelsson E, Jonasson P, Viklund F, Nilsson B, Uhlén M
Department of Biochemistry and Biotechnology, KTH, Stockholm, Sweden.
Nat Biotechnol. 1996 Jun;14(6):751-5. doi: 10.1038/nbt0696-751.
We show that coexpression of a specific binding protein in Escherichia coli can significantly improve the relative yields of correctly folded human insulin-like growth factor I (IGF-I). A glutathione redox buffer was used during growth to allow formation and breakage of disulfide bonds in the periplasm of the bacterial host. Both the binding protein and the peptide hormone were produced as affinity fusions, which allowed purification of the in vivo formed heterodimer by alternative affinity purification methods. The use of affinity-assisted in vivo folding has general implications for expression, folding, and purification of recombinant proteins.
我们发现，在大肠杆菌中共表达一种特定的结合蛋白能够显著提高正确折叠的人胰岛素样生长因子I（IGF-I）的相对产量。在细菌宿主生长过程中使用了谷胱甘肽氧化还原缓冲液，以使周质中的二硫键能够形成和断裂。结合蛋白和肽激素均作为亲和融合蛋白产生，这使得能够通过交替亲和纯化方法纯化体内形成的异二聚体。亲和辅助体内折叠的应用对重组蛋白的表达、折叠和纯化具有普遍意义。
