Submission Title

Rescue Mutations for an X-Prolyl Dipeptidyl Aminopeptidase (PEPX) W425G Variant

Presenter Information

Cody Kaiser, Whitworth University

Session Number

PS1

Location

Graves Gym

Abstract Number

PS1-k

Abstract

Celiac disease is an autoimmune disease that is caused by an allergic response to the gluten ingested in food. Modern treatment mainly consists of avoidance of dietary gluten but potentially oral treatment using enzyme therapy could be developed. One such enzyme is PEPX from Lactobacillus helveticus. Initial studies using a W425G PEPX variant from L. helveticus, designed for pepsin resistance, exhibited instability. Three rescue mutations were made to stabilize the PEPX variant and hopefully retain activity. Mutations L424A, F230A, and I433A were successfully made by site-directed mutagenesis to help rescue the W425G. However activity assays showed that the rescue mutations did not provide the stability needed to retain activity. Further testing is needed to conclusively determine the mechanism of activity loss.

This document is currently not available here.

COinS
 
Apr 23rd, 10:45 AM Apr 23rd, 12:15 PM

Rescue Mutations for an X-Prolyl Dipeptidyl Aminopeptidase (PEPX) W425G Variant

Graves Gym

Celiac disease is an autoimmune disease that is caused by an allergic response to the gluten ingested in food. Modern treatment mainly consists of avoidance of dietary gluten but potentially oral treatment using enzyme therapy could be developed. One such enzyme is PEPX from Lactobacillus helveticus. Initial studies using a W425G PEPX variant from L. helveticus, designed for pepsin resistance, exhibited instability. Three rescue mutations were made to stabilize the PEPX variant and hopefully retain activity. Mutations L424A, F230A, and I433A were successfully made by site-directed mutagenesis to help rescue the W425G. However activity assays showed that the rescue mutations did not provide the stability needed to retain activity. Further testing is needed to conclusively determine the mechanism of activity loss.