Purification of polygalacturonase from solid-state cultures of Aspergillus carbonarius.
Purification of polygalacturonase (PG) from the cultures of Aspergillus carbonarius obtained by acetate buffer extraction after solid-state fermentation was attempted by integrated membrane process and alginate affinity precipitation. The carbohydrates were completely eliminated (98%-99%) with a PG recovery of 72%-80% during integrated membrane process, which would otherwise interfere with the purification process and lead to enzyme loss. However, specific activity of PG did not improve (1.19-1.21 fold) due to the presence of other similar molecular mass proteins. Under optimum conditions of affinity precipitation, the specific activity of PG enhanced to 2450 U/mg (4 fold) with almost complete elimination of carbohydrates and colour compounds resulting in a PG recovery of 61%. PG purity obtained with ultrafiltration (UF) was comparable with the conventional dialysis during desalting eluted PG, besides UF rendered a concentrated PG. The enzyme purity stated was as descend by SDS-PAGE. The results suggested suitability of affinity precipitation for PG purification from solid-state cultures and the potential of UF as a single step process for handling eluted PG.
Department of Food Engineering, Central Food Technological Research Institute, Council of Scientific and Industrial Research, Mysore, India.
Ekambaram
E
Nakkeeran
Rangaswamy
R
Subramanian
Sukumaran
S
Umesh-Kumar
D001230:Aspergillus / Q000201:enzymology*; D004591:Electrophoresis, Polyacrylamide Gel; D005590:Fractional Precipitation; D008954:Models, Biological; D011096:Polygalacturonase / Q000302:isolation & purification*; D014462:Ultrafiltration
D011096:Polygalacturonase