Avanti Christina, Saluja Vinay, van Streun Erwin L P, Frijlink Henderik W, Hinrichs Wouter L J
Department of Pharmaceutics, Faculty of Pharmacy, University of Surabaya (Ubaya), Surabaya, Indonesia ; Department of Pharmaceutical Technology and Biopharmacy, University of Groningen, Groningen, The Netherlands.
Department of Pharmaceutical Technology and Biopharmacy, University of Groningen, Groningen, The Netherlands ; Pharmaceutical Sciences and Clinical Supply (PSCS), Development Center Oss, MSD, Oss, The Netherlands.
PLoS One. 2014 Jan 23;9(1):e86244. doi: 10.1371/journal.pone.0086244. eCollection 2014.
The purpose of this study was to investigate the stability of lysozyme in aqueous solutions in the presence of various extremolytes (betaine, hydroxyectoine, trehalose, ectoine, and firoin) under different stress conditions. The stability of lysozyme was determined by Nile red Fluorescence Spectroscopy and a bioactivity assay. During heat shock (10 min at 70°C), betaine, trehalose, ectoin and firoin protected lysozyme against inactivation while hydroxyectoine, did not have a significant effect. During accelerated thermal conditions (4 weeks at 55°C), firoin also acted as a stabilizer. In contrast, betaine, hydroxyectoine, trehalose and ectoine destabilized lysozyme under this condition. These findings surprisingly indicate that some extremolytes can stabilize a protein under certain stress conditions but destabilize the same protein under other stress conditions. Therefore it is suggested that for the screening extremolytes to be used for protein stabilization, an appropriate storage conditions should also be taken into account.
本研究的目的是在不同应激条件下，研究溶菌酶在含有各种极端嗜盐菌胞内相容性溶质（甜菜碱、羟基乙磺酸甜菜碱、海藻糖、四氢嘧啶和丝胶蛋白）的水溶液中的稳定性。通过尼罗红荧光光谱法和生物活性测定法来确定溶菌酶的稳定性。在热休克期间（70°C下10分钟），甜菜碱、海藻糖、四氢嘧啶和丝胶蛋白可保护溶菌酶不被灭活，而羟基乙磺酸甜菜碱则没有显著作用。在加速热条件下（55°C下4周），丝胶蛋白也起到了稳定剂的作用。相比之下，在这种条件下，甜菜碱、羟基乙磺酸甜菜碱、海藻糖和四氢嘧啶会使溶菌酶失稳。这些发现令人惊讶地表明，一些极端嗜盐菌胞内相容性溶质在某些应激条件下可以稳定一种蛋白质，但在其他应激条件下却会使同一蛋白质失稳。因此，建议在筛选用于蛋白质稳定化的极端嗜盐菌胞内相容性溶质时，也应考虑适当的储存条件。
