Separation and Property of Chitinase Extracted from Rape Seeds by Chitin Affinity Columns with Three Different Regenerated Chitins

The chitinases of rape seeds were separated by chitin affinity columns regenerated from three different molecular chitosans of 20 cPs, 276 cPs and 1,200 cPs. The chitinase of rape seeds occurred in the order of Ch1, Ch2, and Ch3 on SDS-PAGE. The protein contents recovered were 3.95, 1.59, and 1.90% from chitin affinity columns of 20, 276, and 1200 cPs, while the respective chitinase activities were 47.2, 80.9, and 60.2%. The hydrolysis products of seed chitinase observed on TLC plates primarily consisted of dimers and trimers after 12 hrs of incubation with swollen chitin, while they were primarily dimers and monomers on TLC plates after 12 hrs of incubation with chitin-oligomers. In conclusion, the highest recovery rate of the chitinase activity from rape was obtained using a chitin affinity column filled with 276 cPs chitosan, and the chitinase was a very specific enzyme for the selective production of GlcNAc monomers and dimers.