Abstract
We have developed a NH3/H2O2 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) NH4OH 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) H2O2 (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 NH3/H2O2 two-step method is more practical than the NH3/H2O2 coupling method.
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Acknowledgments
This study was financially supported by the Natural Science Foundation of China (No. 20806057), the Research Fund for the Doctoral Program of Higher Education of China (No. 200800561004), the Ministry of Science and Technology of China (Nos. 2012BAD29B05 and 2012AA06A303), and the Program for New Century Excellent Talents in Chinese University (NCET-11-0372).
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Liu, R., Su, R., Yu, Y. et al. Regeneration of insulin monomers from amyloid fibrils by a NH3/H2O2 two-step method. Biotechnol Lett 34, 1959–1964 (2012). https://doi.org/10.1007/s10529-012-0974-8
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DOI: https://doi.org/10.1007/s10529-012-0974-8