Screening, Purification and Characterization of Pullulanase from Soil Bacteria and Enhancement of Its Stability by Immobilization and Nanoforms

Pullulanase is an extracellular carbohydrase responsible for the hydrolysis of pullulan and amylopectin to
produce maltotriose. The product maltotriose is used in detergent industry, bakery industry and in the
production of biotechnological products. In the present investigation pullulanase was purified from bacillus
species isolated from soil sample. The pullulanase was purified by salt precipitation (70%) followed by anion exchange chromatography (DEAE- Cellulose) and gel filtration (Sephadex G75) with final yield of 75%
and purification fold 16.14. The molecular mass of pullulanase enzyme was 110 kDa as estimated by SDSPAGE. The enzyme activity of purified sample was found to be 1294 IU and specific activity 8271.8. This
study also aimed to determine the stability of immobilized pullulanase and showed high stability using Cu
Nanoparticle.