Purification, Thermostability and Chemical Modification of Pullulanase from Sunflower

eng Excellent Publishers International Journal of Current Microbiology and Applied Sciences (IJCMAS) CODEN(USA)-IJCMO9 2319-7692 2319-7706 2015-12-20 4 12 284 296 article Purification, Thermostability and Chemical Modification of Pullulanase from Sunflower H. M. El-Shora 1 S. E. Ghada 2 H. S. Amany 3 Botany Department, Faculty of Science, Mansoura University, Egypt

Pullulanase (EC 3. 2. 1. 41) activity was measured in sunflower (Helianthus annus L.) cotyledons. The enzyme was purified from sunflower cotyledons with specific activity of 580 units mg-1 protein. The enzyme was purified to homogeneity with single band. The optimal pH 6.0 and the optimal temperature was 45ºC. Sarcosine offered pullulanase appreciable thermostability at 60ºC. The enzyme was modified by various concentrations of dansyl chloride (DnsCl), N-bromosuccinimide (NBS) and N-acetylimidazole (NAI). The modification resulted in an inhibition of enzyme activity with the three tested compounds. The inhibition reveals the presence of lysyl, tryptophenyl and tyrosyl residues. Inclusion of pullulan as a substrate during the incubation offered the enzyme a protection against inactivation.

http://ijcmas.com/vol-4-12/H. M. El-Shora, et al.pdf Pullulanase Sunflower Purification Thermostability