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Please use this identifier to cite or link to this item: http://eprint.iitd.ac.in/handle/2074/1188

Title: Separation of enzymes by sequential macroaffinity ligand-facilitated three-phase partitioning
Authors: Sharma, Aparna
Mondal, Kalyani
Gupta, Munishwar Nath
Keywords: Enzymes
Pectinase
Cellulase
Issue Date: 2003
Citation: Journal of Chromatography A, 995(1-2), 127-134
Abstract: Pectinase and cellulase were separated from a commercial enzyme preparation called Pectinex Ultra SP-L. This was carried out using a process called macroaffinity ligand-facilitated three-phase partitioning (MLFTPP). In this method, a water-soluble polymer is floated as an interfacial precipitate by adding ammonium sulfate and tert.-butanol. The polymer (appropriately chosen) in the presence of an enzyme for which it shows affinity, selectively binds to the enzyme and floats as a polymer–enzyme complex. In the first step, pectinase was purified (with alginate as the polymer) 13-fold with 96% activity recovery. In the second MLFTPP step, using chitosan, cellulase was purified 16-fold with 92% activity recovery. Both preparations showed a single band on sodium dodecylsulfate–polyacrylamide gel electrophoresis. This illustrative example shows that the strategy of sequential MLFTPP can be used to separate important biological activities from a crude broth
URI: http://eprint.iitd.ac.in/dspace/handle/2074/1188
Appears in Collections:Chemistry

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