双水相体系酶纯化：纯聚合物组成的体系与粗聚合物组成的体系之间的比较

Enzyme purification with aqueous two-phase systems: comparison between systems composed of pure polymers and systems composed of crude polymers.

作者信息

Venâncio A, Almeida C, Teixeira J A

机构信息

Centro de Engenharia Biológica, Universidade do Minho, Largo do Paço, Braga, Portugal.

出版信息

J Chromatogr B Biomed Appl. 1996 May 17;680(1-2):131-6. doi: 10.1016/0378-4347(95)00419-x.

DOI:10.1016/0378-4347(95)00419-x

PMID:8798890

Abstract

The main drawback when using aqueous two-phase systems for macromolecule purification is the high cost of most polymers used. The purification of an enzyme, alcohol dehydrogenase, from a crude extract of Saccharomyces cerevisiae was tested in systems composed of poly(ethylene glycol) and a crude hydroxypropyl starch or Reppal PES 100, a purified fraction of hydroxypropyl starch. Purification factors measured for the enzyme were very similar in both systems (between 0.8 and 1.4 for both systems in the upper phase). However, systems composed of Reppal PES present a greater recovery of enzyme, between 77% and 100% versus 60% and 100%, while systems composed of crude hydroxypropyl starch exhibit a larger delta log K for the tested ligand, 1.26 versus 0.81.

摘要

使用双水相系统进行大分子纯化时的主要缺点是所使用的大多数聚合物成本高昂。在由聚乙二醇和粗羟丙基淀粉或Reppal PES 100（羟丙基淀粉的纯化级分）组成的系统中，对从酿酒酵母粗提物中纯化一种酶（乙醇脱氢酶）进行了测试。在两个系统中测得的该酶的纯化因子非常相似（在上相中两个系统均在0.8至1.4之间）。然而，由Reppal PES组成的系统酶回收率更高，为77%至100%，而由粗羟丙基淀粉组成的系统为60%至100%，同时由粗羟丙基淀粉组成的系统对测试配体显示出更大的Δlog K，分别为1.26和0.81。