固定化分子伴侣GroE促进变性溶菌酶复性的研究

利用重组大肠杆菌表达制备了分子伴侣GroE(GroEL和GroES),研究了GroE以及GroEL辅助变性溶菌酶复性的作用。结果表明,不仅游离GroEL单独作用可使溶菌酶复性收率达到90%以上,而且固定化GroEL亦可有效地促进蛋白质复性,最佳复性温度为37℃,最佳pH值范围为6～8,复性酶的活性收率在85%以上。另外,固定化GroEL可反复回收利用,表明固定化GroEL有可能在实际生物下游过程中得到应用。

Reactivation of Denatured Lysozyme with Immobilized Molecular Chaperones GroE

The molecular chaperones GroEL and GroES were expressed in recombinant E. coli and purified by anion exchange chromatography. The renaturation of the denatured lysozyme with the free and immobilized GroEL/ES or GroEL was studied. We show here that using free GroEL alone could reactive the denatured lysozyme up to a relative activity of over 90%. The immobilized GroEL was also effective for promoting lysozyme refolding. Moreover, the optimal temperature (i.e., 37 degrees C) and (pH(i.e., 6 to 8) for the immobilizde GroEL-facilitated lysozyme refolding operation were determined. Under the optimal condition, the activity of lysozyme could be recovered up to 85%. In addition, the immobilized GroEL was repeatedly used five times without loss of its renaturation ability, indicating its potentiality to be used in practical downstream bioprocesses.