Suenaga M, Ohmae H, Tsuji S, Itoh T, Nishimura O
Biotechnology Laboratories, Pharmaceutical Research Division, Takeda Chemical Industries Ltd, Jusohonmachi 2-17-85, Yodogawa-ku, Osaka 532, Japan.
Biotechnol Appl Biochem. 1998 Oct;28 ( Pt 2):119-24.
Escherichia coli has been widely used in the production of recombinant proteins. One of the drawbacks inherent in this method is that the proteins produced in the cells often form inactive inclusion bodies. Usually, the inclusion bodies can be separated from other cell components, solubilized by denaturants such as guanidine hydrochloride or urea, and then renatured through a refolding process such as dilution or dialysis. However, it has been shown that biologically active recombinant human neurotrophin-3 cannot be obtained at high yield by this procedure due to aggregation and precipitation of the protein. We applied the refolding process using the aggregation suppressor L-arginine in the renaturation of neurotrophin-3, and obtained biologically active neurotrophin-3 at high yield from the inclusion bodies. Consequently, about 10 mg of purified neurotrophin-3 was prepared from 1 litre of culture broth.
大肠杆菌已被广泛应用于重组蛋白的生产。这种方法固有的一个缺点是细胞中产生的蛋白质常常形成无活性的包涵体。通常，包涵体可以与其他细胞成分分离，用盐酸胍或尿素等变性剂溶解，然后通过稀释或透析等复性过程进行复性。然而，已经表明，由于该蛋白质的聚集和沉淀，通过此程序无法高产获得具有生物活性的重组人神经营养因子-3。我们在神经营养因子-3的复性过程中应用了使用聚集抑制剂L-精氨酸的复性方法，并从包涵体中高产获得了具有生物活性的神经营养因子-3。因此，从1升培养液中制备了约10毫克纯化的神经营养因子-3。
