A Simple Strategy for the Purification of Large Thermophilic Proteins Overexpressed in Mesophilic Microorganisms: Application to Multimeric Enzymes from Thermus sp. Strain T2 Expressed in Es
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- 作者：Benevides C. C. Pessela ; Rodrigo Torres ; Manuel Fuentes ; Cesar Mateo ; Miguel Filho ; Alfonso V. Carrascosa ; Alejandro Vian ; Jose L. Garcí ; a ; Jose M. Guisá ; n ; and Roberto Fernandez-Lafuente
- 刊名：Biotechnology Progress
- 出版年：2004
- 出版时间：October 2004
- 年：2004
- 卷：20
- 期：5
- 页码：1507 - 1511
- 全文大小：90K
- 年卷期：v.20,no.5(October 2004)
- ISSN：1520-6033
文摘
The heating of protein preparations of mesophilic organism (e.g., E. coli) producesthe obliteration of all soluble multimeric proteins from this organism. In this way, ifa multimeric enzyme from a thermophilic microorganism is expressed in thesemesophilic hosts, the only large protein remaining soluble in the preparation afterheating is the thermophilic enzyme. These large proteins may be then selectivelyadsorbed on lowly activated anionic exchangers, enabling their full purification in justthese two simple steps. This strategy has been applied to the purification of an
-galactosidase and a 
-galactosidase from Thermus sp. strain T2, both expressed inE. coli, achieving the almost full purification of both enzymes in only these two simplesteps. This very simple strategy seems to be of general applicability to the purificationof any thermophilic multimeric enzyme expressed in a mesophilic host.
-galactosidase and a -galactosidase from Thermus sp. strain T2, both expressed inE. coli, achieving the almost full purification of both enzymes in only these two simplesteps. This very simple strategy seems to be of general applicability to the purificationof any thermophilic multimeric enzyme expressed in a mesophilic host.