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Team:UChicago/Project
From 2013.igem.org
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Contents |
Overall project
The goal of our project is to make the tanning industry more environmentally sustainable by making an enzymatically based hide de-hairing system. Ideally, our de-hairing system will involve the expression of a temperature optimized keratinase in B. subtilis. Our experiment is divided into several parts. Expression of keratinase in B. subtilis. First, we isolated the keratinase gene from B. licheniformis by using specially designed primers. Then, we attached constitutive promoter veg to the gene in order to optimize expression. Optimization of keratinase activity. The keratinase protein works optimally at 50ºC; however, heating is cost prohibitive as it means that the protein must first be isolated and purified in large amounts in order to be used for de-hairing. Optimization of protein activity at optimal B. subtilis growth conditions would reduce the cost in time and money of a keratinase based de-hairing system by allowing the B. subtilis to be cultured right on the hide. The strategy we will be using to optimize the protein is directed evolution as not enough is known about the biophysical properties of the protein. Creation of a keratin-dependent B. subtilis strain. We also want to make a B. subtilis strain that can only survive if it has a source of keratin. We would do this by knocking out amino acid synthesis genes for several amino acids that can be obtained from keratin. This way, likelihood of escape into the environment is virtually eliminated.
Project Details
Synthesizing kerA from IDT gBlocks
Expression of KerA in B. subtilis
Optimization of KerA activity
Creation of a Keratin-dependent B. subtilis strain
Results
