Purification of endo-polygalacturonase by affinity precipitation using alginate.
The precipitation of alginate by Ca2+ at pH 3.8 was found to occur concomitantly with the precipitation of endo-polygalacturonase from Aspergillus niger. Under optimum conditions, 92% of the enzyme activity was precipitated. The enzyme could be recovered from the precipitate by washing with 0.5 M NaCl/0.2 M Ca2+. All the precipitated endo-polygalacturonase activity could be recovered in this way. The enzyme thus obtained was purified 10-fold. A comparison of SDS/PAGE gel patterns of the crude preparation and enzyme purified by the affinity precipitation also showed a significant purification of the enzyme.
Gupta
M N
MN
Department of Biotechnology, Lund University, Sweden.
Dong
G
G
Mattiasson
B
B
D000464:Alginates / Q000737:chemistry*; D002118:Calcium / Q000378:metabolism; D011232:Chemical Precipitation; D004591:Electrophoresis, Polyacrylamide Gel; D020723:Glucuronic Acid; D006603:Hexuronic Acids; D006863:Hydrogen-Ion Concentration; D011096:Polygalacturonase / Q000302:isolation & purification* / Q000378:metabolism
D000464:Alginates; D006603:Hexuronic Acids; D020723:Glucuronic Acid; D011096:Polygalacturonase; D002118:Calcium