Use of crosslinked mucilage prepared from ruredzo (Dicerocaryum zanguebarium) in the purification of polygalacturonase extracted from tomato

The mucilage from ruredzo (Dicerocaryum zanguebarium) was crosslinked in alkaline conditions with epichlorohydrin. Polygalacturonase (PG) was extracted from ripe tomatoes by first macerating at pH 3 at low ionic strength. After washing off sugars, the enzyme was extracted in buffer containing 1.7 M NaCl. PG was bound to crosslinked mucilage (CLM) at pH 4 and released by eluting with buffer containing 1 M NaCl after washing off unbound proteins. A six-fold purification of PG was achieved. The purified enzyme showed two main bands of molecular weights 30 000 and 44 000. Used CLM was regenerated by treatment in 1 M NaCl and 1 M HCl. Treatment of PG with regenerated CLM produced results that were similar to the purification with fresh CLM.