Three-phase partitioning for simultaneous renaturation and partial purification of Aspergillus niger xylanase.
Three-phase partitioning (TPP) is carried out by mixing ammonium sulfate and t-butanol to obtain organic phase, interfacial precipitate and aqueous phase. It is shown that TPP of an 8 M urea/100 mM dithiothreitol-denatured xylanase preparation resulted in simultaneous renaturation and purification. This integrated novel approach gave recovery of 93% enzyme activity with 21-fold purification. The implications of this in the context of recovering activity from inclusion bodies are discussed.
Roy
Ipsita
I
Chemistry Department, Indian Institute of Technology, Hauz Khas, New Delhi, 110016, India.
Sharma
Aparna
A
Gupta
Munishwar Nath
MN
D001234:Aspergillus niger / Q000201:enzymology* / Q000378:metabolism; D043364:Endo-1,4-beta Xylanases / Q000302:isolation & purification / Q000378:metabolism*; D007700:Kinetics; D011489:Protein Denaturation; D020673:Protein Renaturation; D014508:Urea
D014508:Urea; D043364:Endo-1,4-beta Xylanases