Simple protein purification through affinity adsorption on regenerated amorphous cellulose followed by intein self-cleavage.
A simple, low-cost, and scalable protein purification method was developed by using a biodegradable regenerated amorphous cellulose (RAC) with a binding capacity of up to 365 mg protein per gram of RAC. The recombinant protein with a cellulose-binding module (CBM) tag can be specifically adsorbed by RAC. In order to avoid using costly protease and simplify purification process, a self-cleavage intein was introduced between CBM and target protein. The cleaved target protein can be liberated from the surface of RAC by intein self-cleavage occurring through a pH change from 8.0 to 6.5. Four recombinant proteins (green fluorescence protein, phosphoglucomutase, cellobiose phosphorylase, and glucan phosphorylase) have been purified successfully.
Hong
Jiong
J
Biological Systems Engineering Department, Virginia Polytechnic Institute and State University, 210-A Seitz Hall, Blacksburg, VA 24061, USA.
Wang
Yiran
Y
Ye
Xinhao
X
Zhang
Y-H Percival
YH
D002482:Cellulose / Q000378:metabolism; D002846:Chromatography, Affinity / Q000379:methods*; D005964:Glucosyltransferases / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D047668:Inteins; D010733:Phosphoglucomutase / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D006005:Phosphorylases / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D011485:Protein Binding; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D013696:Temperature
D011994:Recombinant Proteins; D049452:Green Fluorescent Proteins; D002482:Cellulose; D005964:Glucosyltransferases; D006005:Phosphorylases; C034714:cellobiose phosphorylase; D010733:Phosphoglucomutase