Free polymeric bioligands in aqueous two-phase affinity extractions of microbial xylanases and pullulanase.
Two reversibly soluble-insoluble polymers (viz. Eudragit S-100 and alginate) were used as free macroaffinity bioligands in polyethylene glycol (PEG)/salt two-phase systems for separation of enzymes. Incorporation of Eudragit S-100 and alginate in the PEG phase led to considerable selectivity in separation of microbial xylanases and pullulanase, respectively. Xylanase from Aspergillus niger was recovered 93% with 56-fold purification, whereas the enzyme from Trichoderma reesei and Bacillus amyloliquefaciens was obtained with 93% activity recovery (31-fold purification) and 90% activity recovery (32-fold purification), respectively. From Bacillus acidopullulyticus pullulanase, 85% enzyme activity recovery with 44-fold purification was obtained. The approach described here shows the potential of developing into a general approach for use of reversibly soluble-insoluble macroaffinity ligand in two-phase affinity extraction.
Teotia
S
S
Chemistry Department, Indian Institute of Technology, Hauz Khas, New Delhi 110016, India.
Lata
R
R
Gupta
M N
MN
D000464:Alginates / Q000378:metabolism; D001234:Aspergillus niger / Q000201:enzymology*; D001407:Bacillus / Q000201:enzymology*; D004591:Electrophoresis, Polyacrylamide Gel; D020723:Glucuronic Acid; D006026:Glycoside Hydrolases / Q000302:isolation & purification*; D006603:Hexuronic Acids; D008024:Ligands; D011092:Polyethylene Glycols; D011109:Polymethacrylic Acids / Q000737:chemistry*; D014242:Trichoderma / Q000201:enzymology*; D043325:Xylan Endo-1,3-beta-Xylosidase; D014995:Xylosidases / Q000302:isolation & purification*
D000464:Alginates; D006603:Hexuronic Acids; D008024:Ligands; D011109:Polymethacrylic Acids; C038300:methylmethacrylate-methacrylic acid copolymer; D011092:Polyethylene Glycols; D020723:Glucuronic Acid; D006026:Glycoside Hydrolases; D014995:Xylosidases; D043325:Xylan Endo-1,3-beta-Xylosidase; C100181:pullulanase