在体积排阻色谱柱上研究了还原剂存在时脲和盐酸胍变性的3种溶菌酶溶液的复性和分离过程.当变性液中原始溶菌酶浓度大于10 g/L时,变性溶菌酶在体积排阻色谱柱上除了复性为与未变性溶菌酶出峰时间相同的复性态溶菌酶分子外,还形成了溶菌酶折叠中间体的二分子集聚体.这个结果得到了用稀释法复性时溶菌酶的蛋白电泳检测结果的支持.与稀释法复性相比较,用体积排阻色谱法复性时所形成的折叠中间体二分子集聚体的量要远远低于用稀释法所形成的集聚体的量.
参考文献
| [1] | Ptitsyn O B. Protein Eng, 1994, 7(5): 593 |
| [2] | Hinds D A, Levitt M. Trends Biotechnol, 1995, 13(1): 23 |
| [3] | Gorovits B M, McGee W A, Horowitz P M. Biochim Biophys Acta, 1998, 1382(1): 120 |
| [4] | Rudolph R, Zettlmeissl G, Jaenicke R. Biochemistry, 1979, 18(25): 5572 |
| [5] | Brems D N. Biochemistry, 1988, 27(12): 4541 |
| [6] | Serrano L, Matouschek A, Fersht A R. J Mol Biol, 1992, 224(3): 805 |
| [7] | Hagen A J, Hatton T A, Wang D I C. Biotechnol Bioeng, 1990, 35(10): 966 |
| [8] | London J, Skrzynia C, Goldberg M E. Eur J Biochem, 1974, 47(2): 409 |
| [9] | Goldberg M E, Zetina C R. In: Jaenicke R. Protein Folding. Amsterdam and New York: Elsevier/North-Holland Biomedical Press, 1980. 469 |
| [10] | Batas B, Jones H R, Chaudhuri J B. J Chromatogr A, 1997, 766: 109 |
| [11] | Batas B, Schiraldic C, Chaudhuri J B. J Biotechnology, 1997, 68: 149 |
| [12] | Batas B, Chaudhuri J B. Biotechnol Bioeng, 1996, 50(1): 16 |
| [13] | Lu Diannan, Liu Zheng. Journal of Chemical Industry and Engineering (China) (卢滇楠, 刘铮. 化工学报), 2002, 53(10): 1028 |
| [14] | Ristow S S, Wetlaufer D B. Biochem Biophys Res Commun, 1973, 50(2): 544 |
| [15] | Wetlaufer D B, Saxena V P. Biochemistry, 1970, 9(25): 5015 |
| [16] | Dudley K S, Lars J L, Dwight E N, Ingemar B. Biochemistry, 1991, 30(11): 2797 |
| [17] | Chang J Y, Li L. FEBS Lett, 2002, 511: 73 |
| [18] | Sasahara K, Demura M, Nitta K. Proteins, 2002, 49: 472 |
| [19] | Yang Fang, Liang Yi, Yang Fang (Jr). Acta Chimica Sinica (杨芳, 梁毅, 杨芳(小). 化学学报), 2003, 61(6): 803 |
上一张
下一张
上一张
下一张
计量
- 下载量()
- 访问量()
文章评分
- 您的评分：
-
10%
-
20%
-
30%
-
40%
-
50%