Malik Ajamaluddin
Department of Biochemistry, Protein Research Chair, College of Science, King Saud University, PO Box 2455, Riyadh, 11451, Saudi Arabia.
3 Biotech. 2016 Jun;6(1):44. doi: 10.1007/s13205-016-0397-7. Epub 2016 Feb 4.
Disulfide bonds occurred in majority of secreted protein. Formation of correct disulfide bonds are must for achieving native conformation, solubility and activity. Production of recombinant proteins containing disulfide bond for therapeutic, diagnostic and various other purposes is a challenging task of research. Production of such proteins in the reducing cytosolic compartment of E. coli usually ends up in inclusion bodies formation. Refolding of inclusion bodies can be difficult, time and labor consuming and uneconomical. Translocation of these proteins into the oxidative periplasmic compartment provides correct environment to undergo proper disulfide bonds formation and thus achieving native conformation. However, not all proteins can be efficiently translocated to the periplasm with the help of bacterial signal peptides. Therefore, fusion to a small well-folded and stable periplasmic protein is more promising for periplasmic production of disulfide bonded proteins. In the past decades, several full-length proteins or domains were used for enhancing translocation and solubility. Here, protein fusion tags that significantly increase the yields of target proteins in the periplasmic space are reviewed.
大多数分泌蛋白中都存在二硫键。形成正确的二硫键是实现天然构象、溶解性和活性所必需的。为了治疗、诊断和其他各种目的而生产含有二硫键的重组蛋白是一项具有挑战性的研究任务。在大肠杆菌还原性的胞质区室中生产此类蛋白通常最终会形成包涵体。包涵体的重折叠可能很困难，耗时耗力且不经济。将这些蛋白转运到氧化性的周质区室可提供正确的环境，使其形成适当的二硫键，从而实现天然构象。然而，并非所有蛋白都能借助细菌信号肽有效地转运到周质中。因此，与一个折叠良好且稳定的小周质蛋白融合对于在周质中生产二硫键结合蛋白更具前景。在过去几十年中，几种全长蛋白或结构域被用于提高转运和溶解性。在此，对能显著提高周质空间中靶蛋白产量的蛋白融合标签进行综述。
