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三相分配法作为一种从甘薯块茎(Solanum tuberosum)中快速简便地纯化和回收过氧化氢酶的方法。

Three-phase partitioning as a rapid and easy method for the purification and recovery of catalase from sweet potato tubers (Solanum tuberosum).

机构信息

Section of Biochemistry, Department of Chemistry, Faculty of Arts and Sciences, Kocaeli University, Umuttepe Central Campus, 41380 İzmit, Kocaeli, Turkey.

出版信息

Appl Biochem Biotechnol. 2013 Jul;170(5):1119-26. doi: 10.1007/s12010-013-0260-9. Epub 2013 May 3.

Abstract

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.

摘要

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。

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