[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.
Dong
X Y
XY
Department of Biochemical Engineering, Tianjin University.
Yang
H
H
Gan
Y R
YR
Bai
S
S
Sun
Y
Y
D018835:Chaperonin 10 / Q000494:pharmacology*; D018834:Chaperonin 60 / Q000494:pharmacology*; D004789:Enzyme Activation; D009113:Muramidase / Q000737:chemistry*; D011489:Protein Denaturation; D017510:Protein Folding
D018835:Chaperonin 10; D018834:Chaperonin 60; D009113:Muramidase