Institute of Biochemistry, Heinrich-Heine-Universität Düsseldorf, Düsseldorf, Germany.
Bioengineered. 2012 Sep-Oct;3(5):289-92. doi: 10.4161/bioe.20712. Epub 2012 Jun 29.
Protein production through dedicated secretion systems might offer an potential alternative to the conventional cytoplasmical expression. The application of Type 1 secretion systems of Gram-negative bacteria, however, where often not successful in the past for a wide range of proteins. Recently, two studies using the E. coli maltose binding protein (MalE) and the rat intestinal fatty acid binding protein (IFABP) revealed a rational to circumvent these limitations. Here, wild-type passenger proteins were not secreted, while folding mutants with decreased folding kinetics were efficiently exported to the extracellular space. Subsequently, an one-step purification protocol yielded homogeneous and active protein. Taken together, theses two studies suggest that the introduction of slow-folding mutations into a protein sequence might be the key to use Type 1 secretion systems for the biotechnological production of proteins.
通过专用分泌系统进行蛋白质生产可能提供了一种替代传统细胞质表达的潜在方法。然而，革兰氏阴性菌的 I 型分泌系统的应用在过去对于广泛的蛋白质来说并不成功。最近，两项使用大肠杆菌麦芽糖结合蛋白（MalE）和大鼠肠脂肪酸结合蛋白（IFABP）的研究揭示了规避这些限制的合理方法。在这里，野生型载体蛋白没有被分泌，而折叠动力学降低的折叠突变体则被有效地输出到细胞外空间。随后，通过一步纯化方案得到了均一和活性的蛋白质。总之，这两项研究表明，在蛋白质序列中引入缓慢折叠突变可能是利用 I 型分泌系统进行蛋白质生物技术生产的关键。
