Macroaffinity ligand-facilitated three-phase partitioning (MLFTPP) for purification of xylanase

✍ Scribed by Aparna Sharma; M. N. Gupta

• Book ID: 101724436
• Publisher: John Wiley and Sons
• Year: 2002
• Tongue: English
• Weight: 118 KB
• Volume: 80
• Category: Article
• ISSN: 0006-3592
• DOI: 10.1002/bit.10364

✦ Synopsis

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).