Team:U of Guelph/Description
Project Description
Beerstone, FRC and OXC
Calcium oxalate build up (commonly referred to as Beerstone) is a major ongoing problem in the brewing industry. Beerstone is difficult to remove from brewing equipment, requiring the use of harsh cleaning chemicals, and provides a porous, nutrient rich growing environment for bacterial biofilms. Our project is to develop a biological, enzyme based cleaning method for the removal of Beerstone from brewing equipment. We propose to use the enzymes FRC and OXC from Oxalobacter formigenes oxalate degrading metabolic pathway to breakdown Beerstone.
Project Overview
Step 1: Cloning of frc and oxc into DH5α
-Synthesize frc and oxc
-Add PstI cut site to pET-28a
-Ligate frc and oxc into pET-28a
-Transform pET-28afrc/oxc into DH5α
Step 2: Clone frc and oxc into BL21
-Purify pET-28afrc/oxc from DH5α
-Transform pET-28afrc/oxc into BL21
Step 3: Express and Purify FRC and OXC
-Induce expression with IPTG and extract crude proteins
-Purify proteins using Ni-NTA chromatography
Step 4: Characterize FRC and OXC
-Characterize enzyme function using Sodium Oxalate
-Characterize enzyme function using Calcium Oxalate
Step 5: Design a Cleaning Solution and Test on Beerstone
-Test ability of enzymes to break down Beerstone
-Design a functional cleaning solution
The 2017 Aspect
Due to setbacks and unforeseen circumstances our team was only able to accomplish Step 1 this year. We will however be continuing our project next year where we left off. This winter we will be re-grouping in an effort to better plan for next year so that our project execution occurs more smoothly and we are able to accomplish more.
