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Please use this identifier to cite or link to this item: http://eprint.iitd.ac.in/handle/2074/1146

Title: Macroaffinity ligand-facilitated three-phase partitioning for purification of glucoamylase and pullulanase using alginate
Authors: Mondal, Kalyani
Sharma, Aparna
Gupta, Munishwar Nath
Keywords: Alginates
Smart polymers
Three-phase partitioning (TPP)
Macroaffinity ligand-facilitated three-phase partitioning (MLFTPP)
Issue Date: 2003
Citation: Protein Expression and Purification, 28(1), 190-195
Abstract: Starch-degrading enzymes glucoamylase (from Aspergillus niger), and pullulanase (from Bacillus acidopullulyticus) were purified using alginates (polysaccharides consisting of mannuronic acids and guluronic acids) by a recently developed technique called macroaffinity ligand-facilitated three-phase partitioning (MLFTPP). In this process, a crude preparation of the enzyme was mixed with alginate. On addition of appropriate amounts of ammonium sulfate and t-butanol, the alginate bound enzyme appeared as an interfacial precipitate between the lower aqueous and the upper t-butanol phase. Enzyme activity from this interfacial precipitate was recovered using 1 M maltose. Glucoamylase and pullulanase were purified 20- and 38-fold with 83% and 89% activity recovery, respectively. Both the purified preparations showed a single band on SDS–PAGE.
URI: http://eprint.iitd.ac.in/dspace/handle/2074/1146
Appears in Collections:Chemistry

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