Institute of Biomaterials and Bioengineering, Tokyo Medical and Dental University, Tokyo, Japan.
Macromol Biosci. 2011 Jun 14;11(6):814-20. doi: 10.1002/mabi.201000457. Epub 2011 Mar 7.
Cell-free protein synthesis is a promising technique for the rapid production of proteins. However, the application of the cell-free systems requires the development of an artificial chaperone that prevents aggregation of the protein and supports its correct folding. Here, nanogel-based artificial chaperones are introduced that improve the folding efficiency of rhodanese produced in cell-free systems. Although rhodanese suffers from rapid aggregation, rhodanese was successfully expressed in the presence of the nanogel and folded to the enzymatically active form after addition of cyclodextrin. To validate the general applicability, the cell-free synthesis of ten water-soluble proteins was examined. It is concluded that the nanogel enables efficient expression of proteins with strong aggregation tendency.
无细胞蛋白质合成是一种很有前途的蛋白质快速生产技术。然而，无细胞系统的应用需要开发一种人工伴侣，以防止蛋白质聚集并支持其正确折叠。在这里，引入了基于纳米凝胶的人工伴侣，以提高无细胞系统中产生的硫氰酸酶的折叠效率。尽管硫氰酸酶容易迅速聚集，但在纳米凝胶存在的情况下，硫氰酸酶成功表达，并在添加环糊精后折叠成具有酶活性的形式。为了验证其普遍适用性，考察了十种水溶性蛋白质的无细胞合成。结论是，纳米凝胶能够有效地表达具有强烈聚集趋势的蛋白质。
