Hamada Hiroyuki, Shiraki Kentaro
Institute of Applied Physics, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki 305-8573, Japan.
J Biotechnol. 2007 Jun 15;130(2):153-60. doi: 10.1016/j.jbiotec.2007.03.003. Epub 2007 Mar 12.
L-arginine (Arg) is a widely used additive for suppressing protein aggregation during refolding. Systematic screening of Arg analogs provides superior additives that enhance the refolding yield more effectively than Arg. The refolding yield of hen egg lysozyme in the presence of 500 mM L-argininamide (ArgAd) increases 1.7-fold higher than Arg. Thermal unfolding experiments indicate that ArgAd has a greater denaturing effect than Arg. The refolding yield positively relates to the net charge of Arg analogs. Moreover ArgAd was also effective for the refolding of bovine carbonic anhydrase. High potency to increase the refolding yield of ArgAd compared to Arg results from high positive net charge and the denaturing property.
L-精氨酸(Arg)是一种在重折叠过程中广泛用于抑制蛋白质聚集的添加剂。对精氨酸类似物进行系统筛选可得到比精氨酸更有效地提高重折叠产率的优质添加剂。在500 mM L-精氨酰胺(ArgAd)存在的情况下,鸡蛋清溶菌酶的重折叠产率比精氨酸提高了1.7倍。热变性实验表明,ArgAd比精氨酸具有更大的变性作用。重折叠产率与精氨酸类似物的净电荷呈正相关。此外,ArgAd对牛碳酸酐酶的重折叠也有效。与精氨酸相比,ArgAd提高重折叠产率的高效性源于其高正净电荷和变性特性。