Loading...
Confirming the Mannanase Function of Protein Structure 3CBW
Journal Title
Keywords
Readers/Advisors
Koeppe, Julia
Journal Title
Term and Year
Publication Date
2025
Book Title
Publication Volume
Publication Issue
Publication Begin
Publication End
Number of pages
Collections
Files
Loading...
Quest2025_044.pdf
Adobe PDF, 413.88 KB
Research Projects
Organizational Units
Journal Issue
Abstract
This project aimed to verify the mannanase activity of 3CBW, an enzyme with a known structure but unknown function. Expression is done by adding plasmid DNA containing the gene for 3CBW to E. coli to perform transformation. Enzyme 3CBW is purified from lysed E. coli cells by Ni-NTA chromatography. Kinetic assays confirm the computational predictions made prior to testing, which are performed using Azcl-galactomannan (AZCL-GM), a substrate that is hydrolyzed by endo-1,4-β-mannanases. This enzyme successfully hydrolyzes this substrate, measured by the formation of a soluble blue product from the insoluble substrate.