Yasuda M, Murakami Y, Sowa A, Ogino H, Ishikawa H
Department of Chemical Engineering, Osaka Prefecture University, 1-1 Gakuen-cho, Sakai, Osaka 599-8531, Japan.
Biotechnol Prog. 1998 Jul-Aug;14(4):601-6. doi: 10.1021/bp9800438.
Denatured lysozyme was refolded by a dilution method. The refolding yield depended greatly on the lysozyme concentration in the refolding mixture. When the concentration of denatured lysozyme was 0.02 g/L, the refolding yield was as high as 60%. However, when the concentration of denatured lysozyme was 0.2 g/L, the refolding yield was as low as 10% due to the formation of aggregates. To prevent the formation of aggregates and to increase the refolding yield at a low cost, inexpensive additives were screened. The addition of acetone, acetoamide, or urea derivatives was very effective for improving the refolding yield. To clarify why the addition of acetoamide in the refolding mixture improved the refolding yield at the high lysozyme concentration, the time courses of the concentration and the average diameter of the aggregates in the refolding mixture were monitored by the dynamic light scattering method. The experimental results showed that acetoamide played a role in preventing the formation and growth of aggregates and secondary aggregation between the lysozyme aggregates.
变性溶菌酶通过稀释法进行复性。复性产率在很大程度上取决于复性混合物中溶菌酶的浓度。当变性溶菌酶的浓度为0.02 g/L时，复性产率高达60%。然而，当变性溶菌酶的浓度为0.2 g/L时，由于聚集体的形成，复性产率低至10%。为了防止聚集体的形成并以低成本提高复性产率，筛选了廉价的添加剂。添加丙酮、乙酰胺或尿素衍生物对提高复性产率非常有效。为了阐明为什么在复性混合物中添加乙酰胺能在高溶菌酶浓度下提高复性产率，通过动态光散射法监测了复性混合物中聚集体的浓度和平均直径随时间的变化过程。实验结果表明，乙酰胺在防止聚集体的形成和生长以及溶菌酶聚集体之间的二次聚集方面发挥了作用。
