Macroaffinity ligand-facilitated three-phase partitioning (MLFTPP) for purification of xylanase |
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Authors: | Sharma Aparna Gupta M N |
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Affiliation: | Chemistry Department, Indian Institute of Technology, Hauz Khas, New Delhi 110016, India. |
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Abstract: | It is shown that eudragit S-100, a copolymer of methylacrylic acid and methylmethacrylate, undergoes three-phase partitioning. It was found that 95% eudragit S-100 could be recovered as the interfacial precipitate by using 30% (w/v) ammonium sulfate, 1:1 ratio of t-butanol to polymer solution at 40 degrees C. Three-phase partitioning of proteins uses simultaneous addition of ammonium sulfate and t-butanol to precipitate proteins in an interfacial layer separating the aqueous phase and organic solvent. Exploiting the affinity of xylanases towards eudragit S-100, it was possible to purify xylanase from Aspergillus niger; 60% recovery of activity with 95-fold purification could be obtained by this process. The purified enzyme showed A single band on SDS-PAGE. The technique shows promise to develop into a general method that could be termed "macroaffinity ligand-facilitated three-phase partitioning (MLFTPP). |
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Keywords: | eudragit S‐100 protein separation xylanases three phase partitioning |
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