Purification of Pectin Lyase Enzyme from Bacillus pumilus Bacteria by Three-Phase Partitioning Method (TPP), Nanoflower Preparation and Investigation of Fruit Juice Clarification

Abstract

In this study, firstly, Bacillus pumilus bacteria were isolated from tomato vegetables and identified. Then, the new pectin lyase enzyme was purified from B. pumilus, characterized, and hybrid nanoflower pectin lyase (hNF-PL) was synthesized the first time in this study. For this purpose, PL enzyme was produced in a solid culture medium using B. pumilus bacterium, and PL was purified in 191.8 folds with a yield of 78.2% using the three-phase partitioning (TPP) technique. Using SDS-PAGE, PL enzyme was determined to have a single subunit, and molecular weight was defined as 32.88 kDa with gel chromatography technique. This is the very first study to easily immobilize purified PL enzyme onto nanoflower chitosan/calcium pyrophosphate hybrid NPs. The synthesized nanoflower hNF-PL structure was characterized by SEM, FT-IR, XRD, and TEM chromatographic methods. In the final phase of the study, the effects of the pure PL and hNF-PL enzymes on the clarification and cleavage rate of fruit juices obtained from black grape, pomegranate, peach, red apple, and plum were investigated. Under the light shed by this study determined that the hNF-PL enzyme clarified the fruit juices more effectively than the pure PL enzyme.