Purification of Alpha-Amylase in Red Pitaya

Presenter: Danielle Bruner

Co-Presenter(s):
Katherine Flynn, Valerie Crawford

Presenter Status: Undergraduate student

Academic Year: 20-21

Semester: Spring

Faculty Mentor: Monica Lares

Department: Chemistry

Funding Source/Sponsor: Class Project

President's Strategic Plan Goal: Sustainability and Environmental Inquiry

Screenshot URL: https://drive.google.com/uc?id=1YQ7Nv2Ypf52ZnSV1p22o_U0YIkB4uoV8

Abstract:
While α-amylase is usually purified from bacterial and fungal sources in industrial uses, we sought to understand how it could be purified from a plant source, Hylocereus polyrhizus (red pitaya). Our study consists of three protein purification techniques, ammonium sulfate precipitation, dialysis, and gel filtration chromatography, to isolate α-amylase from red pitaya peel. We used a 3,5-dinitrosalicylic (DNS) assay and Bradford assay to measure the enzymatic activity and protein content of our purification samples. Assays were performed after dialysis due to ammonium sulfate being removed and no longer interfering with our enzyme activity. Overall, specific activity increased by the final purification step, however total activity also increased. Further experiments should be performed before industrial use.