Faculty of Pure and Applied Sciences, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki 305-8573, Japan.
Nanomaterials Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), 1-1-1 Higashi, Tsukuba, Ibaraki 305-8565, Japan.
Int J Biol Macromol. 2018 Nov;119:180-185. doi: 10.1016/j.ijbiomac.2018.07.043. Epub 2018 Jul 26.
Allantoin is used to suppress protein aggregation without decreasing the melting temperature. However, the solubility of allantoin in water or buffer solutions is too low (approximately 30 mM at ambient temperature) to be used as a protein aggregation suppressor in various situations. Here we show that a high-concentration solution of allantoin in neat dimethyl sulfoxide (DMSO) is useful as a stock solution for the additive that controls protein aggregation. The allantoin concentration from 10 to 100 mM in 10% (v/v) DMSO significantly suppressed the thermal aggregation of hen egg white lysozyme as a model protein, with increasing allantoin concentration. The residual activity of lysozyme in 10% DMSO and 100 mM allantoin after heating at 90 °C for 10 min was increased >3-fold compared to that without allantoin. Thus, it was concluded that allantoin in DMSO is an effective stock solution for practical application in enhancing the recovery of enzymatic activity and suppressing the formation of small aggregate of protein.
尿囊素在不降低熔点的情况下用于抑制蛋白质聚集。然而，尿囊素在水中或缓冲溶液中的溶解度太低（在环境温度下约为 30mM），无法在各种情况下用作蛋白质聚集抑制剂。在这里，我们表明，高浓度的尿囊素在纯二甲亚砜（DMSO）中的溶液可用作控制蛋白质聚集的添加剂的储备溶液。10%（v/v）DMSO 中浓度为 10 至 100mM 的尿囊素显著抑制了作为模型蛋白的鸡卵清白溶菌酶的热聚集，随着尿囊素浓度的增加而增加。与不含尿囊素相比，在 90°C 加热 10 分钟后，在 10%DMSO 和 100mM 尿囊素中的溶菌酶的残留活性增加了>3 倍。因此，结论是 DMSO 中的尿囊素是一种有效的储备溶液，可用于提高酶活性的恢复和抑制蛋白质小聚集体的形成。
