Section of Biochemistry, Department of Chemistry, Faculty of Arts and Sciences, Kocaeli University, Umuttepe Central Campus, 41380 İzmit, Kocaeli, Turkey.
Appl Biochem Biotechnol. 2013 Jul;170(5):1119-26. doi: 10.1007/s12010-013-0260-9. Epub 2013 May 3.
Three-phase partitioning (TPP) was used to purify and recover catalase from potato crude extract. The method consists of ammonium sulfate saturation, t-butanol addition, and adjustment of pH, respectively. The best catalase recovery (262 %) and 14.1-fold purification were seen in the interfacial phase in the presence of 40 % (w/v) ammonium sulfate saturation with 1.0:1.0 crude extract/t-butanol ratio (v/v) at pH 7 in a single step. The sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis of the enzyme showed comparatively purification and protein molecular weight was nearly found to be 56 kDa. This study shows that TPP is a simple, economical, and quick method for the recovering of catalase and can be used for the purification process.
采用三相分配法(TPP)从马铃薯粗提物中纯化和回收过氧化氢酶。该方法包括硫酸铵饱和度、叔丁醇添加和 pH 值调节。在 40%(w/v)硫酸铵饱和度、1.0:1.0 粗提物/叔丁醇比(v/v)和 pH 7 的单一相中,最好的过氧化氢酶回收率(262%)和 14.1 倍的纯化度出现在界面相中。酶的十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)分析表明,该方法具有较好的纯化效果,且蛋白质分子量接近 56 kDa。本研究表明,TPP 是一种简单、经济、快速的回收过氧化氢酶的方法,可用于纯化过程。