Cloning and Expressing PEPN in Lactobacillus helveticus
Faculty Sponsor
Deanna Ojennus, dojennus@whitworth.edu
Session Type
Poster Presentation
Research Project Abstract
Celiac disease is an autoimmune disorder triggered by gluten, a protein found in wheat. Currently there is no treatment for celiac disease besides a gluten free diet. This research was directed toward creating a potential treatment for celiac disease by examining bacterial peptidase that could break down gluten. Aminopeptidase N (PEPN) is an enzyme that hydrolyzes N terminal amino acids from a peptide chain. It cuts most amino acids, but it is slow acting at proline residues. The goal of this research was to clone PEPN gene from the bacterium Lactobacillus helveticus to construct a recombinant enzyme for gluten digestion studies. The PEPN gene was amplified by PCR and ligated into a pET-14b E. coli expression plasmid. PEPN expression was induced with IPTG. Overall, the research was successful in cloning and expressing PEPN.
Session Number
PS1
Location
HUB Multipurpose Room
Abstract Number
PS1-d
Cloning and Expressing PEPN in Lactobacillus helveticus
HUB Multipurpose Room
Celiac disease is an autoimmune disorder triggered by gluten, a protein found in wheat. Currently there is no treatment for celiac disease besides a gluten free diet. This research was directed toward creating a potential treatment for celiac disease by examining bacterial peptidase that could break down gluten. Aminopeptidase N (PEPN) is an enzyme that hydrolyzes N terminal amino acids from a peptide chain. It cuts most amino acids, but it is slow acting at proline residues. The goal of this research was to clone PEPN gene from the bacterium Lactobacillus helveticus to construct a recombinant enzyme for gluten digestion studies. The PEPN gene was amplified by PCR and ligated into a pET-14b E. coli expression plasmid. PEPN expression was induced with IPTG. Overall, the research was successful in cloning and expressing PEPN.
