Difference between revisions of "Part:BBa K590022"

Revision as of 23:24, 24 September 2011

Kumamolisin-As_G319S, D358G, D368H

A mutated Kumamolisin-As enzyme aimed to break down gluten by increased activity with the PQLP peptide, an antigenic epitope in gliadin.

Usage and Biology

This part was constructed by the [http://2011.igem.org/Team:Washington 2011 University of Washington] iGEM team to break down gluten, the primary cause of Celiac's disease. To test BBa _K590021, it was inserted into a protein expression vector, pET29b+. . Kumamolisin-As_G319S, D358G, D368H was then produced and purified as described in the [http://2011.igem.org/Team:Washington/Protocols/50mL_UW 2011 iGEM Team's Small Scale Protein Expression and Purification Protocol]. The purified protein was then tested for activity. For a detailed description of the assay, please see the [http://2011.igem.org/Team:Washington/Protocols/Purified_Enzyme_Assay 2011 UW iGEM Purified Enzyme Assay Protocol]. The resulting data is shown below.

The relative activity plot was produced by measuring the quantity of PQLP peptide cleaved per enzyme per hour. Error bars represent a 95% confidence interval from triplicate data.

Sequence and Features