Regeneration of insulin monomers from amyloid fibrils by a NH3/H2O2 two-step method.
We have developed a NH(3)/H(2)O(2) two-step method for the recovery of insulin monomers from amyloid fibrils by modulating the cleavage and regeneration of disulfide bonds. Insulin fibrils were disaggregated into insulin A- and B-chains in 14 M (w/v) NH(4)OH for 2 h at 60 °C. Insulin monomers, with a MW of ~5,882 Da, were then regenerated by oxidation of sulfhydryls with 30 % (w/v) H(2)O(2) (10 M) for 12 h at 25 °C. No two A-chains or two B-chains of insulin formed during the oxidation process. Because of the inconformity of the optimal reduction and oxidation temperature, the NH(3)/H(2)O(2) two-step method is more practical than the NH(3)/H(2)O(2) coupling method.
State Key Laboratory of Chemical Engineering, School of Chemical Engineering and Technology, Tianjin University, 92 Weijin Road, Nankai District, Tianjin 300072, China.
Rui
R
Liu
Rongxin
R
Su
Yanjun
Y
Yu
Wei
W
Qi
Libing
L
Wang
Zhimin
Z
He
D000641:Ammonia / Q000737:chemistry* / Q000494:pharmacology; D064753:Ammonium Hydroxide; D000682:Amyloid / Q000737:chemistry* / Q000187:drug effects / Q000378:metabolism; D000818:Animals; D002417:Cattle; D006861:Hydrogen Peroxide / Q000737:chemistry* / Q000494:pharmacology; D006878:Hydroxides / Q000737:chemistry / Q000494:pharmacology; D007328:Insulin / Q000737:chemistry* / Q000378:metabolism; D058849:Protein Refolding / Q000187:drug effects
D000682:Amyloid; D006878:Hydroxides; D007328:Insulin; D064753:Ammonium Hydroxide; D000641:Ammonia; D006861:Hydrogen Peroxide