PROTEIN EXTRACTION AND ACTIVITY IN REVERSE MICELLES OF A NONIONIC DETERGENT

被引:66
作者
AYALA, GA
KAMAT, S
BECKMAN, EJ
RUSSELL, AJ
机构
[1] UNIV PITTSBURGH,DEPT CHEM ENGN,PITTSBURGH,PA 15261
[2] UNIV PITTSBURGH,CTR BIOTECHNOL & BIOENGN,PITTSBURGH,PA 15261
关键词
ENZYMES; PROTEINS; ORGANIC SOLVENTS; MICROEMULSIONS; NONIONIC SURFACTANTS;
D O I
10.1002/bit.260390803
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
We describe, for the first time, the ability of a polyoxyethylene sorbitan trioleate-isopropanol microemulsion in hexane to solubilize pure proteins. The dependences of cytochrome c extraction and buffer solubilization by the reverse micellar system on ionic strength of the aqueous phase, detergent concentration, and cosurfactant concentration are described. Increases in ionic strength result in decreased recovery of protein in the organic phase, whereas increases in detergent concentration and cosurfactant concentration result in increased extraction. In addition, subtilisin (a serine protease) is shown to be active in this microemulsion. Further the activity of the enzyme can be regulated by the water content of the micelles, enabling control of enzyme activity by "solvent engineering."
引用
收藏
页码:806 / 814
页数:9
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