Abstract
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.
Publication types
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Research Support, U.S. Gov't, Non-P.H.S.
MeSH terms
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Cellulose / metabolism
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Chromatography, Affinity / methods*
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Glucosyltransferases / genetics
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Glucosyltransferases / isolation & purification
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Glucosyltransferases / metabolism
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Green Fluorescent Proteins / genetics
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Green Fluorescent Proteins / isolation & purification
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Green Fluorescent Proteins / metabolism
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Hydrogen-Ion Concentration
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Inteins*
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Phosphoglucomutase / genetics
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Phosphoglucomutase / isolation & purification
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Phosphoglucomutase / metabolism
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Phosphorylases / genetics
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Phosphorylases / isolation & purification
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Phosphorylases / metabolism
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Protein Binding
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Recombinant Proteins / genetics
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Recombinant Proteins / isolation & purification*
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Recombinant Proteins / metabolism
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Temperature
Substances
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Recombinant Proteins
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Green Fluorescent Proteins
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Cellulose
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Glucosyltransferases
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Phosphorylases
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cellobiose phosphorylase
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Phosphoglucomutase